研究者詳細 - 岡野　栄之

論文 - 岡野　栄之

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Disorders in myelination in the twitcher mutant

Mikoshiba Katsuhiko, Fujishiro Masatoshi, Kohsaka Shinichi, Okano Hideyuki, Takamatsu Ken, Tsukada Yasuzo

Neurochemical Research 10 （ 8 ） 1129 - 1141 1985年08月

ISSN 0364-3190

The twitcher is an autosomal recessive mutant mouse characterized by absence of galactosylceramidase. The twitcher shows clinical and histological features similar to those of human Krabbe-type leukodystrophy. We here present the results of a neurochemical and immunohistochemical analysis of the twitcher. Electrophoretic analysis revealed that in the particulate fraction of the spinal cord, myelin basic proteins (MBP) and proteolipid protein were decreased, and in the sciatic nerve fibers, PO protein, X, Y and MBP were clearly decreased. 2′,3′-Cyclic nucleotide 3′-phosphodiesterase (CNPase) activities of the pallium cerebri, brain stem and spinal cord of the twitcher were about 20% less than those of the control. However, in the sciatic nerve, the activity was half that of the control. Immunohistochemical studies were carried out by means of antisera against MBP and CNPase. There were clear patches indicating both MBP- and CNPase-negative reactions in the white matter of the central nervous system from the twitcher. The reaction on the section of sciatic nerve fibers from the twitcher showed a positive reaction only in a very limited number of fibers with both MBP and CNPase antisera. A clear astrocytic hypertrophy was detected by the antiserum against glial fibrillary acidic protein (GFAP). Even in the grey matter of the cerebral cortex, strong GFAP-positive astrocytes were clearly observed. © 1985 Plenum Publishing Corporation.

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Molecular genetic analysis of myelin-deficient mice: shiverer mutant mice show deletion in gene(s) coding for myelin basic protein.

1) Kimura M, Inoko H, Katsuki M, Ando, A, Sato T, Hirose T, Takashima H, Inayama S, Okano H, Takamatsu K, Mikoshiba K, Tsukada Y, Watanabe I.

Journal of Neurochemistry 44 692 - 696 1985年

研究論文（学術雑誌）, 共著, 査読有り

The gene expression of myelin basic proteins (MBPs) in shiverer mutant mice was investigated by the Northern and Southern hybridization techniques. In the control mice RNA molecules from the brains which were about 2,300 nucleotides in length were hybridized to cDNA of 1.8 kb encoding for a mouse MBP, but RNA from the brains of 3-week-old shiverer mutant mice contained no detectable amount of MBP transcripts hybridizing to this probe. Moreover the shiverer mutant mice lost several restriction fragments that hybridized to the same probe in the control mice when each of the five restriction enzymes, i.e., HindIII, PstI, PvuII, AccI, and StuI, was used. These data suggest that the shiverer mutation may correspond to the deletion of a large portion of MBP exon(s) in the gene, and this deletion causes inefficient transcription leading to the depletion of MBPs in the myelin and the dysmyelination observed in these mice.

P₄₀₀ protein characteristic to Purkinje cells and related proteins in cerebella from neuropathological mutant mice

Mikoshiba K., Okano H., Tsukada Y.

Developmental Neuroscience 7 （ 3 ） 179 - 187 1985年

ISSN 0378-5866

P₄₀₀ protein has been found in the cerebellum and localizes to Purkinje cells. It is a glycoprotein and reacts with plant lectins. Autoradiographic patterns of the protein profiles of mutant mice cerebella after intracranial injections of ¹⁴C-leucine were analyzed. P₄₀₀ protein was one of the dominant proteins labeled in the control cerebellum. No incorporation of ¹⁴C-leucine to the position of P₄₀₀ protein was found in the nervous and pcd mutant mice where no Purkinje cells and no P₄₀₀ protein are present. P₄₀₀ protein in weaver cerebellum was labelled more intensely than that in the control. In the staggerer mice, where dendritic arborization of Purkinje cell is poor and no spines are formed on the dendrites, no P₄₀₀ protein was found and no ¹⁴C-leucine incorporation was observed to the position of P₄₀₀ protein, suggesting that staggerer is a mutation where no P₄₀₀ protein accumulated in the fractions being studied. P₄₀₀ protein was phosphorylated independently of the presence of Ca²⁺. Among the proteins of the isolated Purkinje cells, P₄₀₀ protein was one of the major proteins phosphorylated.

Immunohistochemical, biochemical and electron microscopic analysis of myelin formation in the central nervous system of myelin deficient (mld) mutant mice.

Mikoshiba K., Okano H., Inoue Y., Fujishiro M., Takamatsu K., Lachapelle F., Baumann N., Tsukada Y.

Brain Research 432 （ 1 ） 111 - 121 1987年09月

ISSN 0006-8993

Myelin deficiency (mld) is an autosomal recessive mutation in mice and is considered to be allelic to the shiverer (shi) mutation. Mld mice are characterized by hypomyelination of the central nervous system (CNS). They show typical symptoms such as tremor, tonic convulsion and ataxic movement. Subcellular fractionation of the CNS revealed that the MBP bands were greatly decreased in the P2A (myelin) fraction and the total content of myelin basic protein (MBP) was much lower than that in the control in all parts of the CNS. Sections from mld mice were examined by immunohistochemical tests with MBP antiserum, and a mosaic expression of MBP was found in the myelin of the mld mice. Since the major dense line is considered to be composed mainly of MBP, we investigated the myelin of mld mice by electron microscopy and found that there were 3 types of myelin: (1) a normal type compact myelin with a major dense line, (2) a shiverer-type myelin with no major dense line, and (3) a mixed-type myelin, in which within a myelin lamella the major dense line abruptly changes to cytoplasm of oligodendrocytes.

Inefficient transcription of the myelin basic protein gene possibly causes hypomyelination in myelin-deficient mutant mice

Okano H., Miura M., Moriguchi A., Ikenaka K., Tsukada Y., Mikoshiba K.

Journal of Neurochemistry 48 （ 2 ） 470 - 476 1987年

ISSN 0022-3042

A hereditary dysmyelination mutation, named myelin deficient (mld), is considered to be allelic to shiverer, a deletion mutation of the myelin basic protein (MBP) gene. The present study showed that MBP expression is greatly reduced in mld, but that it is still detectable. Northern blot analysis revealed that the pronounced decrease in the MBP level in mld resulted from a reduced mRNA level and was not caused by deletion of a large portion of the MBP gene as in shiverer. Southern blod studies with BamHI-digested chromosomal DNA suggested some part of the MBP gene, at least the 5'-portion, was duplicated in mld. These results indicated that the mld and shiverer mutations were different from each other, even though genetic allelism between the two was reconfirmed. We also examined the developmental pattern of the gene expression of MBP and that of another protein, myelin proteolipid protein (PLP), specifically expressed in the oligodendrocyte, in mld by RNA dot blot study. The mRNA level of MBP in mld was greatly reduced during the active myelination stages, gradually increasing and remaining constant in the later stages. The PLP-mRNA content in mld was almost normal (60-80% that of control) at any stage of development. All these findings imply that the primary defect in mld is due to reduced transcriptional activity of the MBP gene.

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The fifth exon of the myelin proteolipid protein-coding gene is not utilized in the brain of jimpy mutant mice

Moriguchi Akira, Ikenaka Kazuhiro, Furuichi Teiichi, Okano Hideyuki, Iwasaki Yasuno, Mikoshiba Katsuhiko

Gene 55 （ 2-3 ） 333 - 337 1987年

ISSN 0378-1119

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Recombination within the upstream gene of duplicated myelin basic protein genes of myelin deficient shimld mouse results in the production of antisense RNA.

Okano H., Ikenaka K., Mikoshiba K.

EMBO Journal 7 （ 11 ） 3407 - 3412 1988年11月

ISSN 0261-4189

The myelin deficient shimld mouse is an autosomal recessive mutant, characterized by hypomyelination in the central nervous system. The expression of the myelin basic protein (MBP) gene is inhibited transcriptionally. The MBP gene is duplicated tandemly in mld, and exons 3 to 7 of the upstream copy is inverted. In the present studies, we determined the approximate position of the 5' boundary and the nucleotide sequence surrounding the 3' boundary of the inversion and found a number of sequences homologous to the switching regions of mouse immunoglobulin heavy chain gene and J regions of human T cell receptor genes. Antisense RNA complementary to exons 3 and 7, which correspond to the inverted segment, was detected by RNase protection studies. This abnormal transcript was also shown to elongate through the inverted segment to reach the transcription initiation site of the downstream gene.

Central myelin in the first hybrid mice produced by intercrossing homozygotes of shiverer and myelin-deficient mutants

Inoue Yoshiro, Takahashi Shiro, Takayama Chitoshi, Inoue Kaoru, Okano Hideyuki, Mikoshiba Katsuhiko

Brain Research 449 （ 1-2 ） 271 - 280 1988年05月

ISSN 0006-8993

The first hybrid mice ('shiverer*mld' mice) produced by intercrossing the homozygotes of the shiverer (BALB/c strain) and mld (MDB/Dt strain) were used for investigating the fine structure of the myelin lamellae, immunoreactive pattern for myelin basic proteins (MBP) and Golgi impregnated images of oligodendrocytes, with special reference to the influence of aging. All of the hybrid mice had an intermediate coat color between the white of the shiverer and black-brown of the mld, and revealed the same neurological symptoms, intention tremor, ataxic behavior, etc., as those of the shiverer and mld. The central myelin lamellae of the 'shiverer*mld' mouse exhibited the similar characteristics to the shiverer type rather than the mld type from the standpoint of the infrequent occurrence of major dense lines, although they did display a tendency to increase major dense lines with aging like the mld. Observation of the immunohistochemical preparations for MBP showed that immunopositive myelin sheaths were present in the white matter, although they were far more infrequent than those of the mld mutant, probably reflecting the amount of major dense lines. Thus, in the CNS of the 'shiverer*mld' mouse, the MBP-synthesis was possibly much more disturbed than in the mld mutant, or at least, revealed an intermediate pattern between the mld and shiverer. © 1988.

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Myelin proteolipid protein gene structure and its regulation of expression in normal and jimpy mutant mice

Ikenaka Kazuhiro, Furuichi Teiichi, Iwasaki Yasuno, Moriguchi Akira, Okano Hideyuki, Mikoshiba Katsuhiko

Journal of Molecular Biology 199 （ 4 ） 587 - 596 1988年02月

ISSN 0022-2836

The mouse proteolipid protein (PLP) gene was cloned into the λ bacteriophage Charon 4A. The organization and the nucleotide sequence of the exons of the mouse PLP gene were quite similar to those of their human counterparts, consisting of seven exons. The transcription of the PLP gene started from multiple sites. There was a unique sequence tandemly repeated four times, sharing homology with the herpes simplex virus DR2 sequence, upstream from the transcribed region. Expression of the myelin basic protein (MBP) is also restricted to the oligodendrocytes in the central nervous system as is the PLP expression. Homology search against the mouse MBP gene revealed that several boxes in the 5′-flanking region of PLP show a high degree of homology with the sequence present in the MBP 5′-flanking region, possibly of importance in the concomitant expression of both genes in the central nervous system. PLP-mRNA in jimpy mutant mice does not contain exon 5 and its content is greatly reduced. We analyzed the jimpy PLP-mRNA and showed that the transcription initiated from the same sites as those in normal mice. Cloning and sequencing of the 5′-flanking region of the jimpy PLP gene revealed that there were no mutations in the promoter region of the jimpy PLP gene. Therefore, it is likely that a mutation, presumably existing within the jimpy PLP gene, caused the skipping of exon 5 and directly affected the mRNA level. © 1987.

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Gene organization and transcription of duplicated MBP genes of myelin deficient (shi(mld)) mutant mouse.

Okano H., Tamura T., Miura M., Aoyama A., Ikenaka K., Oshimura M., Mikoshiba K.

EMBO Journal 7 （ 1 ） 77 - 83 1988年01月

ISSN 0261-4189

A hereditary dysmyelinating mutation, named myelin deficient (shi(mld)), is characterized by reduced expression of myelin basic protein (MBP). In shi(mld), the MBP gene is duplicated and its reduced expression is mainly determined by the level of mRNA. We have characterized the structure and function of the promoter regions of the duplicated MBP genes in shi(mld). Among the lambda clones containing promoter regions of the duplicated MBP genes in shi(mld), one (gene 1) had the same restriction enzyme pattern as that in control mice, but another (gene 2) had a rearrangement on a distal part of the promoter. A 712-bp nucleotide sequence upstream of the first exons of both of the duplicated MBP genes of shi(mld) was completely consistent with that of the control. Promoter activities of 1.3-kb 5'-flanking regions from respective genes of shi(mld) measured by in vitro run-off assay using HeLa whole-cell extracts were indistinguishable from that of the control MPB gene. Chromosomal mapping by in situ hybridization suggested that the duplicated MBP genes were located closely to each other at the distal part of chromosome 18. A recombinational event including the inversion seemed to have occurred within gene 1 and its possible relationship to the reduced expression of MBP is discussed.

Tissue-specific in vitro transcription from the mouse myelin basic protein promoter

Tamura T. A., Aoyama A., Inoue T., Miura M., Okano H., Mikoshiba K.

Molecular and Cellular Biology 9 （ 7 ） 3122 - 3126 1989年

ISSN 0270-7306

Chimeric and molecular genetic analysis of myelin-deficiency (shiverer and mld) mutant mice

Mikoshiba K., Okano H., Aruga J., Tamura T. A., Miura M., Ikenaka K., Nakagawa T.

Annals of the New York Academy of Sciences 605 166 - 182 1990年

ISSN 0077-8923

The shiverer and myelin-deficient (mld) mutants are two allelic mutations. Both are characterized by hypomyelination in the CNS and deficient expression of the MBP gene. Chimeric analysis of the pathogenesis of shiverer showed that shiverer mutation acts intrinsic to the oligodendrocyte, which is the only cell type expressing. The MBP gene in the CNS. Molecular genetic studies by several groups demonstrated that shiverer is a deletion mutation in the MBP gene. Consequently, no MBP is produced in the shiverer mutant. In mld, however, partial expression of the MBP gene was observed. Interestingly, MBP was expressed in a mosaic fashion in the CNS of mld mice. Molecular genetic studies revealed that the mld mutant has duplicated MBP genes in tandem on a distal part (E2-4) of chromosome 18q. The reduced expression was based on the level of mRNA. A large portion is inverted in the upstream copy, and an intact copy is located downstream. We showed in mld mutants that antisense RNA corresponding to the inverted segment is transcribed, and it forms RNA duplex, with the RNA transcribed from the normal gene located downstream.

Differential localization of alternative spliced transcripts encoding inositol 1,4,5-trisphosphate receptors in mouse cerebellum and hippocampus

Nakagawa Toshiyuki, Shiota Chiyo, Okano Hideyuki, Mikoshiba Katsuhiko

Journal of Neurochemistry 57 （ 5 ） 1807 - 1810 1991年11月

ISSN 0022-3042

Expression of inositol 1,4,5-trisphosphate (InsP₃) receptor subtypes was determined in mouse brain using oligonucleotide-specific in situ hybridization. All subtypes, except one that deletes 120 bp from the full-length InsP₃ receptor cDNA, were expressed in cerebellar Purkinje cells. In hippocampus, various subtypes showed distinct expression patterns. These results suggest that regionally selective expression of InsP₃ receptor subtypes may result in the generation of functionally distinct channels.

Dysmyelination in transgenic mice resulting from expression of class I histocompatibility molecules in oligodendrocytes

Turnley Ann M., Morahan Grant, Okano Hideyuki, Bernard Ora, Mikoshiba Katsuhiko, Allison Janette, Bartlett Perry F., Miller J. F A P

Nature 353 （ 6344 ） 566 - 569 1991年10月

ISSN 0028-0836

MAJOR histocompatibility complex (MHC) molecules are not normally expressed in the central nervous system (CNS)^1-3. However, aberrant expression has been observed in multiple sclerosis lesions and could contribute to the destruction of myelin or the myelinating cells known as oligodendrocytes^4,5. The mechanism of cell damage associated with aberrant MHC molecule expression is unclear: for example, overexpression of class I (ref. 6) and class II (refs 7, 8) MHC molecules in pancreatic β cells in transgenic mice leads to nonimmune destruction of the cells and insulin-dependent diabetes mellitus. We have generated transgenic mice that express class I H-2K^b MHC molecules, under the control of the myelin basic protein promoter, specifically in oligodendrocytes. Homozygous transgenic mice have a shivering phenotype, develop tonic seizures and die at 15-22 days. This phenotype, which we term 'wonky', is due to hypomyelination in the CNS, and not to involvement of the immune system. The primary defect appears to be a shortage of myelinating oligodendrocytes resulting from overexpression of the class I MHC molecules.

The subtypes of the mouse inositol 1,4,5-trisphosphate receptor are expressed in a tissue-specific and developmentally specific manner

Nakagawa Toshiyuki, Okano Hideyuki, Furuichi Teiichi, Aruga Jun, Mikoshiba Katsuhiko

Proceedings of the National Academy of Sciences of the United States of America 88 （ 14 ） 6244 - 6248 1991年07月

ISSN 0027-8424

Additional subtypes of the inositol 1,4,5-trisphosphate (InsP₃) receptor are expressed in a tissue-specific and developmentally specific manner. They differ from the InsP₃ receptor structure previously reported in two small variably spliced segments. One segment (SI) is located within the InsP₃ binding site, whereas another segment (SII) is located near putative sites for phosphorylation and ATP binding to modulate InsP₃ action on Ca²⁺ flux. Therefore, we speculate that selective use of InsP₃ receptor subtypes permits a tissue-specific and developmentally specific expression of functionally distinct channels.

Structure and function of myelin protein genes

Mikoshiba Katsuhiko, Okano Hideyuki, Tamura Taka Aki, Ikenaka Kazuhiro

Annual Review of Neuroscience 14 201 - 217 1991年

ISSN 0147-006X

Identification of the new isoforms of mouse myelin basic protein

Aruga J., Okano H., Mikoshiba K.

Journal of Neurochemistry 56 （ 4 ） 1222 - 1226 1991年

ISSN 0022-3042

Myelin basic protein (MBP) is a major constituent in the myelin of the CNS. In mice, five forms of MBPs (14 kDa, two types of 17 kDa, 18.5 kDa, and 21.5 kDa) encoded by separate mRNAs have been identified based on cDNA cloning studies. These mRNAs are considered to be produced by alternative splicing from a single gene composed of seven exons. Here we report the existence of two novel MBP mRNAs encoding 19.7-kDa and 21-kDa MBPs identified by cDNA cloning using the polymerase chain reaction. Both of these MBPs contain a sequence of a previously unidentified exon of 66 nucleotides, which was mapped to be just 5′ of exon 5 in the MBP gene. MBP mRNAs containing this novel exon (exon 5a) belong to a minor population in the whole brain and PNS and are somewhat enriched in the spinal cord. Exon 5a encodes a very hydrophobic segment rich in valine residues, which presumably forms a β-pleated sheet.

Molecular biology of myelin basic protein

Katsuhiko Mikoshiba, Jun Aruga, Hideyuki Okano

Comparative Biochemistry and Physiology. Part C, Comparative 98 （ 1 ） 51 - 61 1991年

ISSN 0306-4492

1. Myelin is an important structure for facilitating the conduction of impulses along the axons both in the central nervous system (CNS) and peripheral nervous system (PNS). 2. Myelin basic protein (MBP) is a major protein in CNS myelin. 3. MBP is expressed specifically in the nervous system. 4. The MBP gene has been cloned and characterized. 5. Two mutant mice, Shiverer (shi) and myelin-deficient (mld, shi^mid), are autosomal recessive mutants that show severe symptoms such as intentional tremor. They have been found to have a mutation in the MBP gene that results in poor myelination in the central nervous system. 6. It was found that rearrangement within the MBP gene results in low expression of the gene. 7. In Shiverer, the MBP gene is partially deleted (from exons 3 to 7), and in mld, the gene is duplicated tandemly and a large portion of the duplication is inverted upstream of the intact copy. 8. In mld, anti-sense RNA complementary to exons 3-7, which correspond to the inverted segment, was detected by RNase protection studies, and presumed to be responsible for the reduced expressions of MBP. 9.The mechanism of gene rearrangement in MBP was also characterized. 10. This article reviews the recent progress in the study of the MBP gene, especially the rearrangement of the gene and its expression in mutant mice. © 1991.

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Myelin basic protein gene and the function of antisense RNA in its repression in myelin-deficient mutant mouse

Okano Hideyuki, Aruga Jun, Nakagawa Toshiyuki, Shiota Chiyo, Mikoshiba Katsuhiko

Journal of Neurochemistry 56 （ 2 ） 560 - 567 1991年

ISSN 0022-3042

The myelin-deficient (mld) mouse is an autosomal recessive mutant characterized by hypomyelination of the CNS due to reduced expression of the myelin basic protein (MBP) gene. In the mld mutant, the MBP gene is duplicated in tandem. One gene is intact, but a large portion is inverted upstream of the other copy, and its transcription yields the antisense RNA. This antisense RNA was shown to be localized in the nucleus and to form an RNA:RNA duplex with sense RNA. These findings suggested that inhibition of transport from the nucleus or selective degradation of the duplex is responsible for the reduced expression of the MBP gene in the mld mutant. The mechanism of gene rearrangement at the MBP locus was also characterized. Cosmid clones encompassing whole MBP gene loci from control and mld genomic DNA libraries were isolated. The recombination points indicated that the duplication and inversion observed in mld occurred due to nonhomologous recombination.

Recent advances in studies on genes for myelin proteins

Ikenaka K., Okano H., Tamura T. A., Mikoshiba K.

Development Growth and Differentiation 33 （ 3 ） 181 - 192 1991年

ISSN 0012-1592

The inositol 1,4,5-trisphosphate receptor.

Mikoshiba K., Furuichi T., Miyawaki A., Yoshikawa S., Maeda N., Niinobe M., Nakade S., Nakagawa T., Okano H., Aruga J.

Ciba Foundation symposium 164 17 - 29 1992年

ISSN 0300-5208

Inositol 1,4,5-trisphosphate (InsP3) is a second messenger that releases Ca2+ from its intracellular stores. The InsP3 receptor has been purified and its cDNA has been cloned. We have found that the InsP3 receptor is identical to P400 protein, first identified as a protein enriched in cerebellar Purkinje cells. We have generated an L-fibroblast cell transfectant that produces cDNA-derived InsP3 receptors. The protein displays high affinity and specificity for InsP3. InsP3 induces greater Ca2+ release from membrane vesicles from transfected cells than from those from control L-fibroblasts. After incorporation of the purified InsP3 receptor into lipid bilayers InsP3-induced Ca2+ currents were demonstrated. These results suggest that the InsP3 receptor is involved in physiological Ca2+ release. Immunogold labelling using monoclonal antibodies against the receptor showed that it is highly concentrated on the smooth-surfaced endoplasmic reticulum and slightly on the outer nuclear membrane and rough endoplasmic reticulum; no labelling of Golgi apparatus, mitochondria and plasmalemma was seen. Cross-linking experiments showed that the receptor forms a homotetramer. The approximately 650 N-terminal amino acids are highly conserved between mouse and Drosophila, and this region contains the critical sequences for InsP3 binding. We have investigated the heterogeneity of the InsP3 receptor using the polymerase chain reaction and have found novel subtypes of the mouse InsP3 receptor that are expressed in a tissue-specific and developmentally specific manner.

Inositol 1,4,5-trisphosphate (IP3) receptors.

Mikoshiba K., Furuichi T., Miyawaki A., Yoshikawa S., Nakagawa T., Aruga J., Okano H., Yamada N.

Clinical Neuropharmacology 15 Suppl 1 Pt A 1992年

ISSN 0362-5664

The Xenopus IP₃ receptor

Kume Shoen, Muto Akira, Aruga Jun, Nakagawa Toshiyuki, Michikawa Takayuki, Furuichi Teiichi, Nakade Shinji, Okano Hideyuki, Mikoshiba Katsuhiko

Cell 73 （ 3 ） 555 - 570 1993年05月

ISSN 0092-8674

To study the role of the IP₃ receptor (IP₃R) upon egg activation, cDNA clones encoding IP₃R expressed in the Xenopus oocytes were isolated. By analyses of the primary structure and functional expression of the cDNA, Xenopus IP₃R (XIP₃R) was shown to have an IP₃-binding domain and a putative Ca²⁺ channel region. Immunocytochemical studies revealed polarized distribution of XIP₃R in the cytoplasm of the animal hemisphere in a well-organized endoplasmic reticulum-like structure and intensive localization in the perinuclear region of stage VI immature oocytes. In ovulated unfertilized eggs, XIP₃R was densely enriched in the cortical region of both hemispheres in addition to its polarized localization. After fertilization, XIP₃R showed a drastic change in its distribution in the cortical region. These results imply the predominant role of the XIP₃R in both the formation and propagation of Ca²⁺ waves at fertilization. © 1993.

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Isolation of a Drosophila gene encoding a head-specific guanylyl cyclase

Yoshikawa Shingo, Miyamoto Issei, Aruga Jun, Furuichi Teiichi, Okano Hideyuki, Mikoshiba Katsuhiko

Journal of Neurochemistry 60 （ 4 ） 1570 - 1573 1993年04月

ISSN 0022-3042

We have isolated and characterized a new guanylyl cyclase gene (dgc1) in Drosophila. The deduced amino acid sequence (683 amino acids) most closely resembled the mammalian soluble-type guanylyl cyclase α subunit. The cyclase catalytic domain was highly conserved between the mammalian and Drosophila guanylyl cyclases. The dgc1 mRNA was detected in wild-type heads but not in bodies, and its level was reduced in the mutant eyes absent (eya), indicating that dgc1 is preferentially expressed in the CNS and in the eye. The enriched distribution in the eye suggests that dgc1 may have a role in phototransduction.

Regulation of Drosophila neural development by a putative secreted protein

Okano H., Hayashi S., Tanimura T., Sawamoto K., Yoshikawa S., Watanabe J., Iwasaki M., Hirose S., Mikoshiba K., Montell C.

Differentiation 52 （ 1 ） 1 - 11 1993年

ISSN 0301-4681

The Drosophila strawberry (sty) locus was isolated by P-element insertion mutagenesis in a screen for mutations affecting eye development. Analysis of the mutant phenotype and the putative expression pattern of the sty gene suggested that it has multiple functions. Mutations in the sty gene lead to irregular spacing of ommatidia, an increase in the number of photoreceptor cells, as well as abnormal axonal projections to the lamina and disrupted structure of the optic lobes in the adult fly. The sty mutation also causes abnormal head involution, a change in a number of sensilla in the antennomaxillary complex in the embryonic stage and abnormal morphogenesis of the maxillary palp and wings in later stages. We examined the presumptive expression of the sty gene during development by histochemical staining for lacZ expression from enhancer trap elements inserted within the sty gene. During embryogenesis, expression of lacZ showed a segmental pattern in the ectoderm and in the nervous system. In the eye imaginal discs, lacZ was expressed in photoreceptor cells beginning a few rows posterior to the morphogenetic furrow. The lacZ was also expressed in the wing disc. In the adult, lacZ was expressed in the retina and lamina. We cloned the sty gene by P-element tagging and found that it encodes a putative secreted protein containing a cysteine-rich region similar to the epidermal growth factor (EGF) repeat. On the basis of the loss of function phenotype, the expression pattern and the predicted structure of its product, we propose that sty encodes a diffusible protein acting as a signal involved in lateral inhibition within the developing nervous system and also as a factor involved either directly or indirectly in axonal guidance and optic lobe development.

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Structure and function of inositol 1,4,5-trisphosphate receptor

Mikoshiba K., Furuichi T., Miyawaki A., Yoshikawa S., Nakade S., Michikawa T., Nakagawa T., Okano H., Kume S., Muto A., Aruga J., Yamada N., Hamanaka Y., Fujino I., Kobayashi M.

Annals of the New York Academy of Sciences 707 178 - 197 1993年

ISSN 0077-8923

Cloning of a Drosophila melanogaster homologue of the mouse type-I bone morphogenetic proteins-2/-4 receptor

Hideyuki Okano, Shingo Yoshikawa, Atsushi Suzuki, Naoto Ueno, Masahiko Kaizu, Masataka Okabe, Takuya Takahashi, Mineo Matsumoto, Kazunobu Sawamoto, Katsuhiko Mikoshiba

Gene 148 （ 2 ） 203 - 209 1994年10月

ISSN 0378-1119

The Drosophila melanogaster (Dm) decapentaplegic (dpp) gene product plays an essential role during several stages of Dm development. The DPP protein is a member of the transforming growth factor-β (TGF-β) superfamily and an orthologue of mammalian bone morphogenetic proteins (BMP-2 and -4). Recently, a cDNA clone encoding the mouse Ser/Thr kinase receptor specific for BMP-2/-4 (mTFRll) was isolated. Here, we describe the deduced primary structure, the cytogenetic position and expression pattern of the Dm homologue of mTFR11 (DTFR), a putative DPP receptor. The cytogenetic position of the Dm dtfr gene was mapped to 25D. DTFR has striking homology to mTFR11, especially in the cytoplasmic domain (approx. 63%), including a Ser+Gly-rich box that is characteristic of type-I receptors for the TGF-β superfamily. Although the amino acid (aa) sequence of the extracellular domain is less conserved than that of the cytoplasmic domain, the extracellular domains of these two molecules were more homologous (approx. 27%) to each other than any other receptors for the TGF-β superfamily. The spacing of Cys residues in the extracellular domain, which is considered crucial to ligand specificity, is highly conserved in these two receptors. During Dm embryonic development, its expression pattern changes in a dynamic fashion with high levels of expression in mesoderm and midgut, with some relation to dpp mutant phenotypes. © 1994.

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repo encodes a glial-specific homeo domain protein required in the Drosophila nervous system

Xiong Wen Cheng, Okano Hideyuki, Patel Nipam H., Blendy Julie A., Montell Craig

Genes and Development 8 （ 8 ） 981 - 994 1994年04月

ISSN 0890-9369

We report the identification of a Drosophila locus, reversed polarity (repo). Weak repo alleles were viable but affected glia in the optic lobe, resulting in a reversal in polarity of the electrophysiological response to light in the adult. Strong repo alleles caused defects in embryonic glia and resulted in embryonic lethality. Expression of repo appeared to be specific to glia throughout development. In the adult visual system, repo was expressed in laminal glia, medullar glia, and subretinal cells; in the embryo, repo was expressed in nearly all of the identified glia in the central and peripheral nervous systems except midline glia. The repo gene encoded a homeo domain protein suggesting that it might be a transcriptional regulator of genes required for glial development.

The function of argos in regulating cell fate decisions during Drosophila eye and wing vein development

Sawamoto Kazunobu, Okano Hideyuki, Kobayakawa Yoshitaka, Hayashi Shigeo, Mikoshiba Katsuhiko, Tanimura Teiichi

Developmental Biology 164 （ 1 ） 267 - 276 1994年

ISSN 0012-1606

The Drosophila argos gene, which encodes a secreted protein with an EGF motif, is involved in several developmental processes regulating cell-cell interactions such as eye morphogenesis. Loss-of-function mutations in the argos gene cause an increase in the number of photoreceptor cells and cone cells, impaired retinal projections to the optic lobe, and the formation of extra veins. We show here that ubiquitously expressed argos product restored all these loss-of-function phenotypes. Overexpression of argos in the wild- type background resulted in the reduced number of photoreceptor cells, cone cells, and pigment cells, which are phenotypes opposite to those of the loss- of-function mutants. The argos gene is expressed in developing wing veins. Ubiquitous argos expression caused loss of veins in a dose-dependent manner. This phenotype was enhanced by the loss-of-function rhomboid mutation, implying the possibility that argos and rhomboid play key roles in a common pathway for normal wing vein formation. We propose that argos acts as an inhibitory signal for cellular differentiation in the developing eye and wing.

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Musashi, a neural RNA-binding protein required for drosophila adult external sensory organ development

Nakamura Makoto, Okano Hideyuki, Blendy Julie A., Montell Craig

Neuron 13 （ 1 ） 67 - 81 1994年

ISSN 0896-6273

A family of neural RNA-binding proteins has recently been described in both vertebrates and invertebrates. We have identified a new member of this family, the Drosophila musashi (msi) locus, which is required for development of adult external sensory organs (sensilla). In contrast with wild-type sensilla, which contain two outer support cells, the msi mutation typically results in the appearance of extra outer support cells. The msi putative RNA-binding protein is localized to the nucleus and appears to be expressed in all cells in each sensillum and predominantly in neurons during embryogenesis. We propose that the msi protein regulates sensillum development by controlling the expression of target genes at the posttranscriptional level. © 1994.

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Glial cell degeneration and hypomyelination caused by overexpression of myelin proteolipid protein gene

Kagawa Tetsushi, Ikenaka Kazuhiro, Inoue Yoshiro, Kuriyama Shigeki, Tsujii Tadasu, Nakao Junji, Nakajima Kazunori, Aruga Jun, Okano Hideyuki, Mikoshiba Katsuhiko

Neuron 13 （ 2 ） 427 - 442 1994年

ISSN 0896-6273

Myelin proteolipid protein (PLP), the major myelin protein in the CNS, has been thought to function in myelin assembly. Thus, mutations within the gene coding for PLP (Plp) cause hypomyelination, such as the jimpy phenotype in mice and Pelizaeus-Merzbacher disease in humans. However, these mutants often exhibit premature death of oligodendrocytes, which form CNS myelin. To elucidate the functional roles of Pip gene products in the maturation and/or survival of oligodendrocytes, we produced transgenic mice overexpressing the Plp gene by introducing extra wild-type mouse Pip genes. Surprisingly, transgenic mice bearing 4 more Plp genes exhibited dysmyelination in the CNS, whereas those with 2 more Plp genes showed normal myelination at an early age (3 weeks after birth), but later developed demyelination. Overexpression of the Plp gene resulted in arrested maturation of oligodendrocytes, and the severity of arrest was dependent on the extent of overexpression. Overexpression also led to oligodendrocyte cell death, apparently caused by abnormal swelling of the Golgi apparatus. Thus, tight regulation of Plpgene expression is necessary for normal oligodendrocyte differentiation and survival, and its overexpression can be the cause of both dys- and demyelination. © 1994.

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神経系形成の普遍的システムと分岐点

今井眞一郎，岡野栄之

細胞工学 14 （ 12 ） 1434-1443 1995年12月

研究論文（学術雑誌）, 共著

Two major mechanisms regulating cell-fate decisions in the developing nervous system

Okano H.

Development Growth and Differentiation 37 （ 6 ） 619 - 629 1995年

ISSN 0012-1592

Two types of determinants appear to be responsible for the generation of neural cell diversity: non-cell-autonomous and cell-autonomous cues. We have identified both types of determinants through the intensive screening of P-element induced Drosophila mutants affected in neural development. As a member of the first category argos (also referred to as strawberry or giant lens), which regulates cell-cell interaction in the developing nervous system, needs to be mentioned. On the basis of the phenotype on loss of function of argos, its expression pattern and the predicted structure of its product (a secreted protein with a putative epidermal growth factor (EGF) motif) we propose that argos encodes a diffusible protein with pleiotropic functions that acts as a signal involved in lateral inhibition within the developing nervous system and also as a factor involved in axonal guidance. As a member of the second category, I refer to the Drosophila musashi gene that is required for lineage formation and asymmetric division of precursor cells in the developing nervous system. The musashi gene encodes a neural RNA-binding protein and is thus likely to regulate the asymmetric cell division of neural precursor cells by controlling the expression of target genes at the post-transcriptional level. We have identified its mammalian homologue, mouse-musashi-1 (m-Msi-1). In the developing central nervous system (CNS), m-Msi-1 expression was highly enriched to neural precursor cells as is the expression of nestin. Based on the results of a single cell culture experiment, m-Msi-1 expression appears to be associated with multipotent cells that are capable of self-renewal and with the generation of committed precursor cells of both neurons and glia. However, fully differentiated neuronal and glial cells lost their m-Msi-1 expression. The expression of m-Msi protein showed a complementary pattern to that of another mammalian RNA-binding protein Hu, which is localized in differentiated neurons in the CNS. Based on such differential expression patterns and its similarity to the Drosophila musashi, we propose that a combination of neural RNA-binding proteins are required for the asymmetric distribution of intrinsic determinants in the developing mammalian nervous system. The Drosophila glial-specific homeobox protein, Repo, can also be classified as a cell-autonomous cue regulating cell-fate decision during neural development. Repo expression is required for terminal differentiation and for the survival of glial cells.

Molecular genetic analyses of myelin deficiency and cerebellar ataxia

Mikoshiba K., Okano H., Miyawaki A., Furuichi T., Ikenaka K.

Progress in Brain Research 105 23 - 41 1995年

ISSN 0079-6123

Cell-cell interactions during neural development

Sawamoto Kazunobu, Okano Hideyuki

Neuroscience Research 26 （ 3 ） 205 - 214 1996年11月

ISSN 0168-0102

Inhibitory signals of cellular differentiation from differentiating cells play an important role in regulating the number and spatial distribution of distinctive types of cells in developing tissues. Several types of inhibitory mechanisms of cellular differentiation have been identified by making full use of the developmental genetics of Drosophila compound eyes. These inhibitory mechanisms are distinct from each other in their signal transduction cascades and/or their role in the pattern formation of the developing Drosophila eye. The following events occur: firstly a diffusible protein, Scabrous (Sea), is required to confer regular spacings of the founder cells, R8 cells, or preommatidial clusters in the developing eye disc via an unknown signal transduction cascade, secondly the Notch-signalling is at least required for the single-out of the R8 cells within the pre-ommatidial cluster possibly by preventing other cells in the equivalent groups from adapting fates as R8 cells. Notch-signalling activates a simple signal cascade mediating communication between the plasma membrane and nucleus not via protein phosphorylation. In contrast, a novel diffusible ligand, Argos, was likely to be required subsequently to the selection of R8 cells. Argos was shown to inhibit the activation of a receptor tyrosine kinase, DER, and the subsequent signal transduction in the Ras/MAPK cascade (the third inhibitory mechanism). We proposed that the role of Argos is to regulate the number of differentiated cells by controlling cellular differentiation and subsequent programmed cell death. The distinct roles of these inhibitory signals in the developing Drosophila eye are discussed in detail.

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Calcium waves along the cleavage furrows in cleavage-stage Xenopus embryos and its inhibition by heparin

Muto Akira, Kume Shoen, Inoue Takafumi, Okano Hideyuki, Mikoshiba Katsuhiko

Journal of Cell Biology 135 （ 1 ） 181 - 190 1996年10月

ISSN 0021-9525

Calcium signaling is known to be associated with cytokinesis; however, the detailed spatio-temporal pattern of calcium dynamics has remained unclear. We have studied changes of intracellular free calcium in cleavage- stage Xenopus embryos using fluorescent calcium indictor dyes, mainly Calcium Green-1. Cleavage formation was followed by calcium transients that localized to cleavage furrows and propagated along the furrows as calcium waves. The calcium transients at the cleavage furrows were observed at each cleavage furrow at least until blastula stage. The velocity of the calcium waves at the first cleavage furrow was ~3 μ/s, which was much slower than that associated with fertilization/egg activation. These calcium waves traveled only along the cleavage furrows and not in the direction orthogonal to the furrows. These observations imply that there exists an intracellular calcium releasing activity specifically associated with cleavage furrows. The calcium waves occurred in the absence of extracellular calcium and were inhibited in embryos injected with heparin, an inositol 1,4,5-trisphosphate (InsP₃) receptor antagonist. These results suggest that InsP₃ receptor mediated calcium mobilization plays an essential role in calcium wave formation at the cleavage furrows.

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The Drosophila secreted protein Argos regulates signal transduction in the Ras/MAPK pathway

Sawamoto Kazunobu, Okabe Masataka, Tanimura Teiichi, Mikoshiba Katsuhiko, Nishida Yasuyoshi, Okano Hideyuki

Developmental Biology 178 （ 1 ） 13 - 22 1996年08月

ISSN 0012-1606

The Drosophila argos gene encodes a secreted protein with an EGF motif which acts as an inhibitor of cellular differentiation in multiple developmental processes. To investigate the cellular pathways regulated by Argos we screened for mutations which could modify the phenotype caused by overexpression of argos. We show that the effects of argos overexpression on the eye and wing vein development are suppressed by gain-of-function mutations of the MAPKK/D-MEK gene (Dsor1/D-mek) and the MAPK/ERK-A gene (rolled) and were enhanced by loss-of-function mutations of Star. Loss-of-function mutations in components of the Ras/MAPK signaling cascade act as dominant suppressors of the phenotype caused by the argos null mutation. A loss-of-function argos mutation enhanced the overproduction of R7 neurons caused by gain-of-function alleles of Son of sevenless and Dsor1. Conversely, overexpression of argos inhibited formation of the extra R7 cells that was caused by high-level MAPK/ERK-A activity. A phenotype of the sev; argos double mutants revealed that sev is epistatic to argos. These results provide evidence that Argos negatively regulates signal transduction events in the Ras/MAPK cascade.

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Mouse-Musashi-1, a neural RNA-Binding protein highly enriched in the mammalian CNS stem cell

Sakakibara Shin Ichi, Imai Takao, Hamaguchi Kayoko, Okabe Masataka, Aruga Jun, Nakajima Kazunori, Yasutomi Daisuke, Nagata Takashi, Kurihara Yasuyuki, Uesugi Seiichi, Miyata Takaki, Ogawa Masaharu, Mikoshiba Katsuhiko, Okano Hideyuki

Developmental Biology 176 （ 2 ） 230 - 242 1996年06月

ISSN 0012-1606

There is increasing interest in the role of RNA-binding proteins during neural development. Drosophila Musashi is one of the neural RNA-binding proteins essential for neural development and required for asymmetric cell divisions in the Drosophila adult sensory organ development. Here, a novel mammalian neural RNA-binding protein, mouse Musashi-1, was identified based on the homology to Drosophila Musashi and Xenopus NRP-1. In the developing CNS, mouse-Musashi-1 protein was highly enriched in the CNS stem cell. Single-cell culture experiments indicated that mouse Musashi-1 expression is associated with neural precursor cells that are capable of generating neurons and glia. In contrast, in fully differentiated neuronal and glial cells mouse-Musashi-1 expression is lost. This expression pattern of mouse-Musashi-1 is complementary to that of another mammalian neural RNA-binding protein, Hu (a mammalian homologue of a Drosophila neuronal RNA-binding protein Elav), that is expressed in postmitotic neurons within the CNS. In vitro studies indicated that mouse-Musashi-1 possesses binding preferences on poly(G) RNA homopolymer, whereas Hu is known to preferentially bind to short A/U-rich regions in RNA. Based on their differential expression patterns and distinct preferential target RNA sequences, we believe that the mouse-Musashi-1 and Hu proteins may play distinct roles in neurogenesis, either through sequential regulatory mechanisms or differential sorting of mRNA populations during asymmetric division of neural precursor cells.

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The function of the Drosophila argos gene product in the development of embryonic chordotonal organs

Okabe Masataka, Sawamoto Kazunobu, Okano Hideyuki

Developmental Biology 175 （ 1 ） 37 - 49 1996年04月

ISSN 0012-1606

We characterized the embryonic expression pattern and mutant phenotypes of the Drosophila gene argos, which encodes a secreted protein with an epidermal growth factor motif. The argos null mutation caused an increase in chordotonal (Ch) organs in both the thoracic and the abdominal segments, whereas overexpression of the argos gene resulted in a decrease in these organs. We showed that the argos transcripts are expressed transiently in the cells surrounding the Ch organ precursor and that the gene rhomboid (rho), which is involved in the regulation of the number of Ch organs, acts epistatically to argos in this event. Our findings suggest that argos plays a role in Ch organ precursor formation and regulates the final number of Ch organs.

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Ataxia and epileptic seizures in mice lacking type 1 inositol 1,4,5-trisphosphate receptor

Matsumoto M., Nakagawa T., Inoue T., Nagata E., Tanaka K., Takano H., Minowa O., Kuno J., Sakakibara S., Yamada M., Yoneshima H., Miyawaki A., Fukuuchi Y., Furuichi T., Okano H., Mikoshiba K., Noda T.

Nature 379 （ 6561 ） 168 - 171 1996年01月

ISSN 0028-0836

The inositol 1,4,5-trisphosphate (InsP₃) receptor acts as an InsP₃-gated Ca²⁺ release channel in a variety of cell types. Type 1 InsP₃ receptor (IP₃R1) is the major neuronal member of the IP₃R family in the central nervous system, predominantly enriched in cerebellar Purkinje cells but also concentrated in neurons in the hippocampal CA1 region, caudate-putamen, and cerebral cortex. Here we report that most IP₃R1-deficient mice generated by gene targeting die in utero, and born animals have severe ataxia and tonic or tonic-clonic seizures and die by the weaning period. An electroencephalogram showed that they suffer from epilepsy, indicating that IP₃R1 is essential for proper brain function. However, observation by light microscope of the haematoxylin-eosin staining of the brain and peripheral tissues of IP₃R1-deficient mice showed no abnormality, and the unique electrophysiological properties of the cerebellar Purkinje cells of IP₃R1 deficient mice were not severely impaired.

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Augmentation by calmodulin of ADP-ribosylation factor-stimulated phospholipase D activity in permeabilized rabbit peritoneal neutrophils

Takahashi K. I., Tago K., Okano H., Ohya Y., Katada T., Kanaho Y.

Journal of Immunology 156 （ 3 ） 1229 - 1234 1996年

ISSN 0022-1767

Activation of the membrane-bound phospholipase D (PLD) requires cytosolic factor(s), and ADP-ribosylation factor (ARF) has been identified as a cytosolic PLD activator. In the present study, we demonstrate that calmodulin (CaM) and ARF are both involved in PLD activation in rabbit peritoneal neutrophils. The PLD activity of streptolysin O-permeabilized, cytosol- depleted rabbit neutrophils was significantly enhanced when the permeabilized cells were reconstituted with bovine brain cytosol in the presence of guanosine 5'-O-(3-thiotriphosphate) (GTPγS), whereas there was little activation of the enzyme in the absence of cytosol. The GTPγS-stimulated PLD activity in the presence of cytosol was augmented on increasing the concentration of free Ca²⁺. The PLD activity stimulated by GTPγS and Ca²⁺ in this system was inhibited by the calmodulin inhibitor W-7. These findings suggest that CaM plays a role as a cytosolic PLD activator. Moreover, highly purified CaM alone, as well as partially purified ARF alone, promoted a slight stimulation of the PLD activity in permeabilized neutrophils. Interestingly, ARF-stimulated PLD activity was augmented by CaM in the presence of GTPγS and Ca²⁺. This augmentation was again inhibited by W-7, as well as by the structurally unrelated CaM inhibitor trifluoperazine. These data imply that CaM stimulates the PLD activity of rabbit neutrophils in concert with ARF.

hiiragi, a gene essential for wing development in Drosophila melanogaster, affects the Notch cascade

Murata Takehide, Ogura Keiji, Murakami Ryutaro, Okano Hideyuki, Yokoyama Kazunari K.

Genes and Genetic Systems 71 （ 4 ） 247 - 254 1996年

ISSN 1341-7568

A recessive mutation, hiiragi(P1), on the second chromosome of Drosophila was obtained by P element insertion mutagenesis. Flies homozygous for hiiragi(P1) have notched wing margins. Genetic interactions between hiiragi and the genes that encode components of Notch signaling, such as Notch, Hairless, Serrate and deltex, strongly support the involvement of hiiragi in the signal transduction cascade of Notch. It has been reported that Serrate and Delta, other components of Notch signaling, share EGF-like repeats and a second conserved cysteine-rich motif, and that these components interact physically with the same region of Notch. In hiiragi(P1); Serrate(D)/+ double mutants, we observed synergistic enhancement of the notched phenotype of wing margins. In contrast, Delta(FX3) had no phenotypic effect on hiiragi(P1) in hiiragi(P1); Delta(FX3)/+ double mutants. Taken together, these results indicate that hiiragi is involved in the Notch signaling cascade induced by Serrate rather than by Delta.

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argos Is required for projection of photoreceptor axons during optic lobe development in Drosophila

Sawamoto Kazunobu, Okabe Masataka, Tanimura Teiichi, Hayashi Shigeo, Mikoshiba Katsuhiko, Okano Hideyuki

Developmental Dynamics 205 （ 2 ） 162 - 171 1996年

ISSN 1058-8388

The Drosophila argos gene encodes a secreted protein with an epidermal growth factor (EGF) motif, which acts as an inhibitor of cell recruitment in the developing eye and wing. Here, we have analyzed the role of argos during optic lobe development, argos expression was observed in the optic lobes throughout the developmental stages. In argos mutants, neuropiles failed to develop normally during embryonic and larval stages, and photoreceptor axons did not project properly into the lamina. Ubiquitous expression of argos, under control of the hsp70 promoter, rescued the defects in optic lobes. We have found that glial cells failed to differentiate in the larval optic lobes of argos mutants. Correspondingly, in loss-of-function repo mutants, whose glial cells also fail to differentiate, photoreceptor axons showed the impaired projection pattern similar to the argos phenotype. These results suggest that glial cells play a role for guidance of photoreceptor axons. The loss-of-function Star mutation (Star(X155)) dominantly suppressed the defects in the argos optic lobes, suggesting that these two genes act in an antagonistic fashion during optic lobe development.

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Role of inositol 1,4,5-trisphosphate receptor in ventral signaling in xenopus embryos

Kume Shoen, Muto Akira, Inoue Takafumi, Suga Kei, Okano Hideyuki, Mikoshiba Katsuhiko

Science 278 （ 5345 ） 1940 - 1943 1997年12月

ISSN 0036-8075

The inositol 1,4,5-trisphosphate (IP₃) receptor is a calcium ion channel involved in the release of free Ca²⁺ from intracellular stores. For analysis of the role of IP₃-induced Ca²⁺ release (IICR) on patterning of the embryonic body, monoclonal antibodies that inhibit IICR were produced. Injection of these blocking antibodies into the ventral part of early Xenopus embryos induced modest dorsal differentiation. A close correlation between IICR blocking potencies and ectopic dorsal axis induction frequency suggests that an active IP₃-Ca²⁺ signal may participate in the modulation of ventral differentiation.

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Bcl-2 prevents TNF- and Fas-induced cell death but does not inhibit initial processing of caspase-3

Shiraiwa Nobuko, Okano Hideyuki, Miura Masayuki

Biomedical Research 18 （ 6 ） 405 - 411 1997年12月

ISSN 0388-6107

The members of ICE/CED3 family of cysteine proteases (caspases) play important roles in the regulation of programmed cell death. Recent studies have revealed that processing and activation of CPP32 (CASP-3) are crucial for executing cell death in TNF-α- and Fas-induced apoptosis. Overexpression of Bcl-2 in HeLa cells inhibits the cell death and CASP-3 like protease activities which are induced by TNF-α or anti-Fas antibody. Preform of CASP-3 disappered after the treatment with TNF-α or anti-Fas antibody in both HeLa cells and HeLa/bcl-2 cells. These results suggest Bcl-2 prevents cell death and activation of CASP-3 but does not inhibit initial processing of CASP-3.

Recognition of tissue- and subtype- specific modulation of angiotensin II receptors using antibodies against AT1 and AT2 receptors

Rakugi Hiromi, Okamura Atsushi, Kamide Koichi, Ohishi Hiroshi, Sasamura Hiroyuki, Morishita Ryuichi, Higaki Jitsuo, Ogihara Toshio

Hypertens Res 20 （ 5 ） 51-56 1997年12月

研究論文（学術雑誌）, 共著

Evolutionarily conserved mechanisms regulating neural development

Okano H., Okabe M., Taguchi A., Sawamoto K.

Human cell : official journal of Human Cell Research Society 10 （ 3 ） 139 - 150 1997年09月

ISSN 0914-7470

Functions of genes regulating the development of Drosophila peripheral nervous systems are summarized herein. These genes can be classified into 6 groups: <1> prepattern genes, <2> proneural genes, <3> neurogenic genes, <4> neuronal precursor genes, <5> neuronal precursor type selector genes, and <6> cell-division and lineage genes. The mechanisms described herein provide excellent paradigms in the regulation of the development of other tissues in Drosophila, as well as in other organisms, including vertebrates. The roles of two different inhibitory mechanisms, i.e. Notch-signaling and Argos, in the development of Drosophila neural precursor cells are also discussed.

Demonstration of an E-box and its CNS-related binding factors for transcriptional regulation of the mouse type inositol 1,4,5-trisphosphate receptor gene

Konishi Yoshiyuki, Kobayashi Yasushi, Kishimoto Toshihiko, Makino Yasutaka, Miyawaki Atsushi, Furuichi Teiichi, Okano Hideyuki, Mikoshiba Katsuhiko, Tamura Taka Aki

Journal of Neurochemistry 69 （ 2 ） 476 - 484 1997年08月

ISSN 0022-3042

The type 1 inositol 1,4,5-trisphosphate receptor (IP₃R1) is expressed abundantly in the CNS, such as in cerebellar Purkinje cells and the hippocampus. We established a tissue-specific cell-free transcription system and studied regulatory properties of the 5' upstream region of the IP₃R1 gene by use of this system. Deletion analyses of the promoter revealed several cis elements that function significantly in brain nuclear extracts. Among those elements, sequences from -396 to -295 showed the most predominant cerebellum-specific positive function. Footprint analyses demonstrated a factor-binding region from -334 to -318, termed box-I, that contained an E- box consensus sequence. Electrophoretic mobility shift assay revealed CNS- related basic helix-loop-helix proteins for the box-I. Mutational studies using the function assay end competitive electrophoretic mobility shift assays demonstrated a good correlation between the box-I-binding factors and the activated transcription. Box-I-binding factors were present abundantly in adult mouse CNS, whereas their existence was restricted in embryonic and nonneural tissues. Transient chloramphenicol acetyltransferase assay for the IP₃R1 promoter revealed the requirement of box-I in Neuro2a neuroblastoma cells. In the postnatal CNS, multiple basic helix-loop-helix factors are expressed abundantly, some of which are suggested to activate IP₃R1 gene expression in the mammalian CNS.

Developmental expression of the inositol 1,4,5-trisphosphate receptor and localization of inositol 1,4,5-trisphosphate during early embryogenesis in Xenopus laevis

Kume Shoen, Muto Akira, Okano Hideyuki, Mikoshiba Katsuhiko

Mechanisms of Development 66 （ 1-2 ） 157 - 168 1997年08月

ISSN 0925-4773

To study the role of inositol 1,4,5-trisphosphate (IP₃) receptors during early embryogenesis in Xenopus, we examined the temporal-spatial localization of Xenopus IP₃ receptor (XIP3R), XIP3R protein is enriched in the animal hemisphere of early cleavage stage embryos and becomes localized in the ectoderm and involuted mesoderm in gastrula stage embryos. Up to tailbud stages, expression of XIP3R is observed in the mesodermal tissues and in most subregions of the central nervous system. A quantitative analysis of endogenous IP₃ mass during normal early embryogenesis revealed an increase in IP₃ mass first observed at early gastrula stage of 10.5 with an enrichment in the ectoderm throughout the gastrula stages, implying a potential role during gastrulation.

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ICE/CED-3 family executes oligodendrocyte apoptosis by tumor necrosis factor

Hisahara Shin, Shoji ShiN'ichi, Okano Hideyuki, Miura Masayuki

Journal of Neurochemistry 69 （ 1 ） 10 - 20 1997年07月

ISSN 0022-3042

Tumor necrosis factor (TNF) is thought to be one of the mediators responsible for the damage of oligodendrocytes (OLGs) in multiple sclerosis (MS). We report here the involvement of the interleukin 1β-converting enzyme (ICE)/Caenorhabditis elegans gene ced-3 (CED-3) family in TNF-mediated cell death of OLGs. The addition of TNF-α to primary cultures of OLGs that express ice and cpp32 significantly decreased the number of live OLGs in 72 h. DNA fragmentation was detected in TNF-treated OLGs at 36 h with the terminal deoxynucleotidyl transferase dUTP nick end-labeling assay. Benzyloxycarbonyl-Asp-CH ₂OC(O)-2,6-dichlorobenzene, an inhibitor of the ICE/CED-3 family that shows p35-like inhibitory specificity, protected against the TNF-induced cell death of OLGs. Furthermore, acetyl-YVAD-CHO (a specific inhibitor of ICE-like proteases) as well as acetyl-DEVD-CHO (a specific inhibitor of CPP32-like proteases) enhanced the survival of OLGs treated with TNF-α, indicating that ICE- and the CPP32-mediated cell death pathways are activated in TNF-induced OLG cell death. Our results suggest that the inhibition of ICE/CED-3 proteases may be a novel approach to treat neurodegenerative diseases such as MS.

Two-step induction of chordotonal organ precursors in Drosophila embryogenesis

Okabe Masataka, Okano Hideyuki

Development 124 （ 5 ） 1045 - 1053 1997年03月

ISSN 0950-1991

The chordotonal (Ch) organ, an internal stretch receptor located in the subepidermal layer, is one of the major sensory organs in the peripheral nervous system of Drosophila melantogaster. Although the cell lineage of the Ch organ has been well characterized in many studies, the determination machinery of Ch organ precursor cells (COPs) remains largely unresolved. Here we report that the rhomboid (rho) gene and the activity of the Drosophila EGF receptor (DER) signaling pathway are necessary to induce specifically three of the eight COPs in an embryonic abdominal hemisegment. The cell-lineage analysis of COPs using the yeast flpase (flp/FRT) method indicated that each of the eight COPs originated from an individual undifferentiated ectodermal cell. The eight COPs in each abdominal hemisegment seemed to be determined by a two-phase induction: first, five COPs are determined by the action of the proneural gene atonal and neurogenic genes. Subsequently, these five COPs start to express the rho gene, and rho activates the DER-signaling pathway in neighboring cells and induces argos expression. Three of these argos-expressing cells differentiate into the three remaining COPs and they prevent neighboring cells from becoming extra COPs.

Structural properties and RNA-binding activities of two RNA recognition motifs of a mouse neural RNA-binding protein, mouse-Musashi-1

Kurihara Yasuyuki, Nagata Takashi, Imai Takao, Hiwatashi Ado, Horiuchi Masataka, Sakakibara Shin ichi, Katahira Masato, Okano Hideyuki, Uesugi Seiichi

Gene 186 （ 1 ） 21 - 27 1997年02月

ISSN 0378-1119

mouse-Musashi-1 (m-Msi-1) is an RNA-binding protein, abundantly expressed in the developing mammalian central nervous system (CNS). m-Msi-1 contains two RNA recognition motifs (RRMs). In this study, we found that the N-terminal RRM of m-Msi-1 (MMA) binds strongly to poly(G) and weakly to poly(U) in a way similar to that of the full-length m-Msi-1 protein characterized previously. The C-terminal RRM of m-Msi-1 (MMB), however, does not bind to RNA. In addition, the circular dichroism (CD) spectra of the two RRMs showed that the α-helical content of MMA is significantly higher than that of MMB, indicating that some differences in the secondary structure may be responsible for the distinct RNA binding properties of MMA and MMB.

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Developmental expression of the inositol 1,4,5-trisphosphate receptor and structural changes in the endoplasmic reticulum during oogenesis and meiotic maturation of Xenopus laevis

Kume Shoen, Yamamoto Akitsugu, Inoue Takafumi, Muto Akira, Okano Hideyuki, Mikoshiba Katsuhiko

Developmental Biology 182 （ 2 ） 228 - 239 1997年02月

ISSN 0012-1606

To study the development of the calcium release mechanism, we examined the temporal and spatial expression of inositol 1,4,5-trisphosphate receptor (IP3R) during the oogenesis and meiotic maturation of Xenopus laevis. Observation of a series of fixed samples by immunofluorescence microscopy revealed a relocalization of Xenopus IP3R (XIP3R)-positive structures during meiotic maturation. We visualized the endoplasmic reticulum (ER) using ER-sensitive dye, DiI, by observation under confocal laser scanning microscopy. Time-lapse visualization of the living oocytes revealed that while the ER of immature fully grown oocytes underwent relatively little movement, the ER of maturing oocytes and mature eggs moved rapidly. A possible role for the increase of ER mobility in the dynamic redistribution of XIP3R during oocyte maturation is also discussed.

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Intrinsic and extrinsic determinants regulating cell fate decision in developing nervous system

Okabe Masataka, Sawamoto Kazunobu, Imai Takao, Sakakibara Shin Ichi, Yoshikawa Shingo, Okano Hideyuki

Developmental Neuroscience 19 （ 1 ） 9 - 16 1997年01月

ISSN 0378-5866

Cell fate in the nervous system is determined by environmental signals and/or heredity. They can be referred to as extrinsic and intrinsic determinants, respectively. The molecular and cellular mechanisms underlying these processes are well conserved from invertebrates to vertebrates. We have identified both types of determinants through the intensive screening of P-element-induced Drosophila mutants affected in neural development. As a member of the first category, argos gene product which regulates cell-cell interaction in the developing nervous system gives a good example. As a member of the second category, we refer to Drosophila musashi gene encoding a neural RNA-binding protein. The functions of these gene products and application to the mammalian nervous system will be discussed.

Expression of neural RNA-binding proteins in the postnatal CNS

Sakakibara Shin Ichi, Okano Hideyuki

Journal of Neuroscience 17 （ 21 ） 8300 - 8312 1997年

ISSN 0270-6474

There is an increasing interest in the role of RNA-binding proteins during neural development. Mouse-Musashi-1 (m-Msi-1) is a mouse neural RNA- binding protein with sequence similarity to Drosophila musashi (d-msi), which is essential for neural development. m-Msi-1 is highly enriched in neural precursor cells that are capable of generating both neurons and glia during embryonic CNS development. The present study characterized m-Msi-1-expressing cells in the postnatal and adult CNS. Postnatally, m-Msi-1 was expressed in proliferative neuronal precursors in the external granule cell layer of the cerebellum and in the anterior corner of the subventricular zone of the lateral ventricles. In gliogenesis, the persistent expression of m-Msi-1 was observed in cells of the astrocyte lineage ranging from proliferative glial precursors in the subventricular zone (SVZ), to differentiated astrocytes in the parenchyma. In addition, we showed that m-Mis-1 was still expressed in proliferating cells in the adult SVZ, which may contain neural precursor or stem cells. Another neural RNA-binding protein Hu (the mammalian homolog of Drosophila neuronal RNA-binding protein Elav) was present in postmitotic neurons throughout the development of the CNS, and its pattern of expression was compared with that of m-Msi-1. These observations imply that these two RNA-binding proteins may be involved in the development of neurons and glia by regulating gene expression at the post-transcriptional level.

Execution mechanisms of programmed cell death by caspase (ICE/CED-3) family proteases

Miura Masayuki, Hisahara Shin, Araki Takashi, Okano Hideyuki

Heart and Vessels 12 （ SUPPL. 12 ） 66 - 70 1997年

ISSN 0910-8327

Molecular genetic study of Caenorhabditis elegans revealed the cell death-executing gene ced-3. Structural and functional similarities between CED-3 and mammalian ICE family (caspase) gene products provide the evidence that mammalian caspases are common mediators in executing programmed cell death. These findings suggest that the execution mechanisms of cell death by caspase family protease may be evolutionarily conserved from worms to mammals. This report summarizes the unique properties of caspases in apoptosis.

A novel Drosophila paired-like homeobox gene related to Caenorhabditis elegans unc-4 is expressed in subsets of postmitotic neurons and epidermal cells

Tabuchi Katsuhiko, Yoshikawa Shingo, Yuasa Yoshihiro, Sawamoto Kazunobu, Okano Hideyuki

Neuroscience Letters 257 （ 1 ） 49 - 52 1998年11月

ISSN 0304-3940

We have isolated a novel Drosophila paired-like homeobox gene, DPHD-1. The homeodomain of DPHD-1 showed 85% amino-acid identity with that of the C. elegans Unc-4 protein. Whole-mount in situ hybridization of embryos and third-instar larvae revealed that the DPHD-1 mRNA is specifically localized in subsets of postmitotic neurons in the central nervous system (CNS) and in the developing epidermis with a segmentally repeated pattern. Double staining with a posterior compartment marker, an anti-Engrailed antibody, showed that DPHD-1 expressing neurons in the CNS were present in the posterior compartment, whereas DPHD-1 expression in the epidermis was restricted to the anterior compartment in each segment. This temporal and spatial expression pattern suggests that DPHD-1 may play a role in determining the distinct cell types in each segment.

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The human Musashi homolog 1 (MSI1) gene encoding the homologue of Musashi/Nrp-1, a neural RNA-binding protein putatively expressed in CNS stem cells and neural progenitor cells

Good Peter, Yoda Akinori, Sakakibara Shin Ichi, Yamamoto Atsuyo, Imai Takao, Sawa Hitoshi, Ikeuchi Takeshi, Tsuji Shoji, Satoh Hitoshi, Okano Hideyuki

Genomics 52 （ 3 ） 382 - 384 1998年09月

ISSN 0888-7543

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Mammalian BarH homologue is a potential regulator of neural bHLH genes

Saito Tetsuichiro, Sawamoto Kazunobu, Okano Hideyuki, Anderson David J., Mikoshiba Katsuhiko

Developmental Biology 199 （ 2 ） 216 - 225 1998年07月

ISSN 0012-1606

Vertebrate neurogenesis involves sequential actions of transcription factors. neurogenins, encoding Atonal-related bHLH transcription factors, function as neuronal determination genes in Xenopus. neurogenins and another bHLH factor gene, Mash1, are expressed in distinct subsets or areas of cells giving rise to neurons, suggesting that these genes play important roles to generate distinct populations of neurons. A mammalian homologue of BarH (MBH1) is expressed in a complementary pattern to Mash1 expression in the developing nervous system like neurogenins. Forced expression of MBH1 down- regulates expression of Mash1 and up-regulates neurogenin2/Math4A, a member of neurogenins, in P19 cells during neuronal differentiation. This suggests that MBH1 is a potential regulator of mammalian neural bHLH genes, thereby establishing distinct pathways of neuronal differentiation.

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Fibroblast growth factor-2/brain-derived neurotrophic factor-associated maturation of new neurons generated from adult human subependymal cells

Pincus David W., Keyoung H. Michael, Harrison-Restelli Catherine, Goodman Robert R., Fraser Richard A R, Edgar Mark, Sakakibara Shin Ichi, Okano Hideyuki, Nedergaard Maiken, Goldman Steven A.

Annals of Neurology 43 （ 5 ） 576 - 585 1998年05月

ISSN 0364-5134

The adult mammalian forebrain harbors neuronal precursor cells in the subependymal zone (SZ). Neuronal progenitors also persist in the adult human SZ and have been cultured from epileptic temporal lobe. In the present study, we sought to identify these neural progenitors in situ, and to direct their expansion and neuronal differentiation in vitro. We prepared explants of adult human SZ, obtained from temporal lobe resections of refractory epileptics. The resultant cultures were treated with fibroblast growth factor-2 (FGF-2) for a week, with concurrent exposure to [³H]thymidine, then switched to media containing brain-derived neurotrophic factor (BDNF) for up to 2 months. Sporadic neuronal outgrowth, verified antigenically and physiologically, was observed from SZ cultures regardless of FGF-2/BDNF treatment; however, only FGF-2/BDNF-treated cultures exhibited profuse outgrowth, and these displayed neuronal survival as long as 9 weeks in vitro. In addition, cortical cultures derived from two brains generated microtubule- associated protein-2⁺ neurons, which incorporated [³H]thymidine and exhibited significant calcium increments to depolarization. In histological sections of the subependyma, both uncommitted and restricted progenitors, defined respectively by musashi and Hu protein expression, were identified. Thus, the adult human subependyma harbors neural progenitors, which are able to give rise to neurons whose numbers can be supported for prolonged periods in vitro.

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Argos induces programmed cell death in the developing Drosophila eye by inhibition of the Ras pathway

Sawamoto Kazunobu, Taguchi Akiko, Hirota Yuki, Yamada Chiharu, Jin Ming Hao, Okano Hideyuki

Cell Death and Differentiation 5 （ 4 ） 262 - 270 1998年04月

ISSN 1350-9047

We studied the role of Ras signaling in the regulation of cell death during Drosophila eye development. Overexpression of Argos, a diffusible inhibitor of the EGF receptor and Ras signaling, caused excessive cell death in developing eyes at pupal stages. The Argos-induced cell death was suppressed by coexpression of the anti-apoptotic genes p35, diap1, or diap2 in the eye as well as by the Df(3L)H99 chromosomal deletion that lacks three apoptosis-inducing genes, reaper, head involution defective (hid) and grim. Transient misexpression of the activated Ras1 protein (Ras1(V12)) later in pupal development suppressed the Argos-induced cell death. Thus, Argos-induced cell death seemed to have resulted from the suppression of the anti-apoptotic function of Ras. Conversely, cell death induced by overexpression of Hid was suppressed by gain-of-function mutations of the genes coding for MEK and ERK. These results support the idea that Ras signaling functions in two distinct processes during eye development, first triggering the recruitment of cells and later negatively regulating cell death.

Localization of 5-HT2A receptor in rat cerebral cortex and olfactory system revealed by immunohistochemistry using two antibodies raised in rabbit and chicken

Hamada Shun, Senzaki Kouji, Hamaguchi-Hamada Kayoko, Tabuchi Katsuhiko, Yamamoto Hideko, Yamamoto Toshifumi, Yoshikawa Shingo, Okano Hideyuki, Okado Nobuo

Brain Research Molecular Brain Research 54 （ 2 ） 199 - 211 1998年03月

ISSN 0169-328X

Serotonin 2A receptor (5-HT2A receptor) is widely distributed in the central nervous system, and has been suggested to be involved in a variety of behavioral conditions and neuropsychiatric disorders. Two polyclonal antibodies were raised against the N-terminus peptide of rat 5-HT2A receptor in chickens (5-HT2A-N) and a glutathione S-transferase fusion protein that contained the C-terminus of the mouse 5-HT2A receptor in rabbits (5-HT2A-C). Affinity-purified 5-HT2A-N and -C antibodies reacted strongly with a single band of 77-78 kDa in postsynaptic density proteins prepared from the rat cortex. The distribution pattern of immunoreactive structures in the rat brain was virtually the same for the two antibodies. The highest levels of immunoreactivity were observed in the olfactory bulb, neocortex, claustrum, piriform cortex, mamillary bodies, pontine nuclei, red nucleus and cranial motor nuclei. In the olfactory bulb, mitral cells were intensely labeled. In the neocortex, many immunoreactive neurons were found in layers II-VI. In layer IV of the neocortex, strong neuropil labeling was observed. In a double-labeling study using chicken 5-HT2A-N and rabbit anti-glial fibrillary acidic protein (GFAP) antibody, a considerable number of GFAP positive cells also showed 5-HT2A immunoreactivity. By using an immunoelectron microscopic technique, 5-HT2A receptor immunoreaction was shown to be localized just beneath the postsynaptic membrane thickening of asymmetric synapses.

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Inositol 1,4,5-trisphosphate receptor mediates ventralising signals in Xenopus embryos

Kume Shoen, Muto Akira, Okano Hideyuki, Mikoshiba Katsuhiko

Zygote 6 （ 1 SUPPL. ） 1998年

ISSN 0967-1994

Caenorhabditis elegans anti-apoptotic gene ced-9 prevents ced-3-induced cell death in Drosophila cells

Hisahara Shin, Kanuka Hirotaka, Shoji Shin Ichi, Yoshikawa Shingo, Okano Hideyuki, Miura Masayuki

Journal of Cell Science 111 （ 6 ） 667 - 673 1998年

ISSN 0021-9533

ced-9, a member of the bcl-2 gene family in Caenorhabditis elegans plays a central roles in preventing cell death in worms. Overexpression of human bcl-2 can partially prevent cell death in C. elegans. However, it remains to be elucidated whether ced-9 can regulate cell death when expressed in other organisms. We demonstrated that the CED-9 protein is co-localized with BCL-2 in COS cells and Drosophila Schneider's L2 (SL2) cells, suggesting that the site of CED-9 action is located to specific cytoplasmic compartments. Overexpression of ced-9 only poorly protected cells from the death induced by ced-3 in HeLa cells, but ced-9 significantly reduced the cell death induced by ced-3 in Drosophila SL2 cells. Furthermore, apoptosis of SL2 cells that was induced by a Drosophila cell-death gene, reaper, was shown to be partially prevented by ced-9, bcl-2 and bcl-xL. These results suggest that the signaling pathway that is required for the anti-apoptotic function of bcl-2 family members, including ced-9, is conserved in Drosophila cells. In addition, SL2 cells provide a unique systems for dissecting the main machinery of cell death.

Control of the cell death pathway by Dapaf-1, a Drosophila Apaf-1/CED-4-related caspase activator

Kanuka Hirotaka, Sawamoto Kazunobu, Inohara Naohiro, Matsuno Kenji, Okano Hideyuki, Miura Masayuki

Molecular Cell 4 （ 5 ） 757 - 769 1999年11月

ISSN 1097-2765

We identified a Drosophila Apaf-1/CED-4 homolog gene, dapaf-1. Alternative splicing results in two dapaf-1 mRNA species, which encode distinct forms of caspase activator, Dapaf-1L (Apaf-1 type) and Dapaf-1S (CED-4 type). Distinct caspases were activated by these Dapaf-1 isoforms. Loss of Dapaf-1 function resulted in defective cytochrome c-dependent caspase activities and reduced apoptosis in embryo and in larval brain. Dapaf-1 activities were also involved in cell death induced by ectopic expression of reaper in the compound eye. These data suggest that Dapaf-1/cytochrome c-dependent cell death-inducing machinery is present in Drosophila, and the requirement of Dapaf-1/Apaf-1 in neural cell death is conserved through evolution.

Cloning and characterization of Dfak56, a homolog of focal adhesion kinase, in Drosophila melanogaster

Fujimoto Jiro, Sawamoto Kazunobu, Okabe Masataka, Takagi Yasumitsu, Tezuka Tohru, Yoshikawa Shingo, Ryo Haruko, Okano Hideyuki, Yamamoto Tadashi

Journal of Biological Chemistry 274 （ 41 ） 29196 - 29201 1999年10月

ISSN 0021-9258

The focal adhesion kinase (FAK) protein-tyrosine kinase plays important roles in cell adhesion in vertebrates. Using polymerase chain reaction-based cloning strategy, we cloned a Drosophila gene that is homologous to the vertebrate FAK family of protein-tyrosine kinases. We designated this gene Dfak56 and characterized its gene product. The overall protein structure and deduced amino acid sequence of Dfak56 show significant similarity to those of FAK and PYK2. Dfak56 has in vitro autophosphorylation activity at tyrosine residues. Expression of the Dfak56 mRNA and the protein was observed in the central nervous system and the muscle-epidermis attachment site in the embryo, where Drosophila position-specific integrins are localized. The results suggest that like FAK in vertebrates, Dfak56 functions downstream of integrins. Dfak56 was tyrosine-phosphorylated upon integrin-dependent attachment of the cell to the extracellular matrix. We conclude that the Dfak56 tyrosine kinase is involved in integrin-mediated cell adhesion signaling and thus is a functional homolog of vertebrate FAK.

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Musashi and Seven in absentia downregulate Tramtrack through distinct mechanisms in Drosophila eye development

Hirota Yuki, Okabe Masataka, Imai Takao, Kurusu Mitsuhiko, Yamamoto Atsuyo, Miyao Sachiyo, Nakamura Makoto, Sawamoto Kazunobu, Okano Hideyuki

Mechanisms of Development 87 （ 1-2 ） 93 - 101 1999年09月

ISSN 0925-4773

We have examined the roles played by the Drosophila neural RNA-binding protein Musashi (MSI) in eye development. MSI expression was observed in the nuclei of all photoreceptor cells (R1-R8). Although a msi loss-of-function mutation resulted in only weak abnormalities in photoreceptor differentiation, we found that the msi eye phenotype was significantly enhanced in a seven in absentia (sina) background. sina is known to be involved in the degradation of the Tramtrack (TTK) protein, leading to the specification of the R7 fate. We demonstrated that MSI also functions to regulate TTK expression. The sina msi mutants showed significantly high ectopic expression of TTK69 and failure in the determination of the R1, R6, and R7 fates. Other photoreceptor cells also failed to differentiate with abnormalities occurring late in the differentiation process. These results suggest that MSI and SINA function redundantly to downregulate TTK in developing photoreceptor cells. Copyright (C) 1999 Elsevier Science Ireland Ltd.

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Mammalian ELAV-like neuronal RNA-binding proteins HuB and HuC promote neuronal development in both the central and the peripheral nervous systems

Akamatsu Wado, Okano Hirotaka J., Osumi Noriko, Inoue Takayoshi, Nakamura Shun, Sakakibara Shin Ichi, Miura Masayuki, Matsuo Nobutake, Darnell Robert B., Okano Hideyuki

Proceedings of the National Academy of Sciences of the United States of America 96 （ 17 ） 9885 - 9890 1999年08月

ISSN 0027-8424

Hu proteins are mammalian embryonic lethal abnormal visual system (ELAV)-like neuronal RNA-binding proteins that contain three RNA recognition motifs. Although Drosophila ELAV is required for the correct differentiation and survival of neurons, the roles played by the Hu genes in the mammalian nervous system remain largely unknown. To explore the in vivo functions of mouse Hu proteins, we overexpressed them in rat pheochromocytoma PC12 cells, where they induced neuronal phenotype in the absence of nerve growth factor. We have characterized the functions of various forms of mHuB and mHuC bearing point mutations or deletions. Mutants of mHuC that had amino acid exchanges in the RNP1 domain of the first or second RNA recognition motifs (RRMs) lost biologic activity as well as RNA-binding activity. In addition, the mutants containing only the third RRM failed to induce the neuronal phenotype in PC12 cells and inhibited the biologic activity of cotransfected wild-type mHuB and mHuC, thus acting as a dominant-negative form. However, these mutants could not suppress the nerve growth factor-induced differentiation of PC12 cells. Further, we misexpressed wild-type and dominant-negative Hu in E9.5 mouse embryos, by using electroporation into the neural tube at the level of the rhombencephalon, mHuB and mHuC induced the ectopic expression of neuronal markers, whereas the dominant-negative forms of mHuB and mHuC suppressed the differentiation of central nervous system motor neurons. From these results, we suggest that Hu proteins are required for neuronal differentiation in the mammalian nervous system.

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The Drosophila Ral GTPase regulates developmental cell shape changes through the Jun NH₂-terminal kinase pathway

Sawamoto Kazunobu, Winge Per, Koyama Shinya, Hirota Yuki, Yamada Chiharu, Miyao Sachiyo, Yoshikawa Shingo, Jin Ming Hao, Kikuchi Akira, Okano Hideyuki

Journal of Cell Biology 146 （ 2 ） 361 - 372 1999年07月

ISSN 0021-9525

The Ral GTPase is activated by RalGDS, which is one of the effector proteins for Ras. Previous studies have suggested that Ral might function to regulate the cytoskeleton; however, its in vivo function is unknown. We have identified a Drosophila homologue of Ral that is widely expressed during embryogenesis and imaginal disc development. Two mutant Drosophila Ral (DRal) proteins, DRal(G20V) and DRal(S25N), were generated and analyzed for nucleotide binding and GTPase activity. The biochemical analyses demonstrated that DRal(G20V) and DRal(S25N) act as constitutively active and dominant negative mutants, respectively. Overexpression of the wild-type DRal did not cause any visible phenotype, whereas DRal(G20V) and DRal(S25N) mutants caused defects in the development of various tissues including the cuticular surface, which is covered by parallel arrays of polarized structures such as hairs and sensory bristles. The dominant negative DRal protein caused defects in the development of hairs and bristles. These phenotypes were genetically suppressed by loss of function mutations of hemipterous and basket, encoding Drosophila Jun NH₂-terminal kinase kinase (JNKK) and Jun NH₂-terminal kinase (JNK), respectively. Expression of the constitutively active DRal protein caused defects in the process of dorsal closure during embryogenesis and inhibited the phosphorylation of JNK in cultured S2 cells. These results indicate that DRal regulates developmental cell shape changes through the JNK pathway.

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Caspase activity is required for nephrogenesis in the developing mouse metanephros

Araki Takashi, Saruta Takao, Okano Hideyuki, Miura Masayuki

Experimental Cell Research 248 （ 2 ） 423 - 429 1999年05月

ISSN 0014-4827

Programmed cell death is a mechanism through which organisms get rid of unwanted cells and is thought to be an important process in organogenesis. Although large-scale cell death is observed in the developing kidney, the precise roles of cell death in kidney organogenesis remain to be elucidated. To address this question, we prevented cell death in metanephric explants by applying caspase inhibitors. Administration of caspase inhibitors (Z-D- CH₂DCB and Ac-DEVD-CHO) effectively prevented the cell death that is normally observed in nondifferentiating mesenchymal cells. Both ureteric bud branching and nephrogenesis were prevented by caspase inhibition. Our results suggest that caspases are crucial in kidney organogenesis and cell death in the nondifferentiating mesenchyme.

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Structure, backbone dynamics and interactions with RNA of the C-terminal RNA-binding domain of a mouse neural RNA-binding protein, Musashi1

Nagata Takashi, Kanno Reisuke, Kurihara Yasuyuki, Uesugi Seiichi, Imai Takao, Sakakibara Shin ichi, Okano Hideyuki, Katahira Masato

Journal of Molecular Biology 287 （ 2 ） 315 - 330 1999年03月

ISSN 0022-2836

Musashi1 is an RNA-binding protein abundantly expressed in the developing mouse central nervous system. Its restricted expression in neural precursor cells suggests that it is involved in the regulation of asymmetric cell division. Musashi1 contains two ribonucleoprotein (RNP)-type RNA-binding domains (RBDs), RBD1 and RBD2. Our previous studies showed that RBD1 alone binds to RNA, while the binding of RBD2 is not detected under the same conditions. Joining of RBD2 to RBD1, however, increases the affinity to greater than that of RBD1 alone, indicating that RBD2 contributes to RNA-binding. We have determined the three-dimensional solution structure of the C-terminal RBD (RBD2) of Musashi1 by NMR. It folds into a compact up structure comprising a four-stranded antiparallel β-sheet packed against two α-helices, which is characteristic of RNP-type RBDs. Special structural features of RBD2 include a β-bulge in β2 and a shallow twist of the β-sheet. The smaller ¹H-¹⁵N nuclear Overhauser enhancement values for the residues of loop 3 between β2 and β3 suggest that this loop is flexible in the time-scale of nano- to picosecond order. The smaller ¹⁵N T₂ values for the residues around the border between α2 and the following loop (loop 5) suggest this region undergoes conformational exchange in the milli- to microsecond time-scale. Chemical shift perturbation analysis indicated that RBD2 binds to an RNA oligomer obtained by in vitro selection under the conditions for NMR measurements, and thus the nature of the weak RNA-binding of RBD2 was successfully characterized by NMR, which is otherwise difficult to assess. Mainly the residues of the surface composed of the four-stranded β-sheet, loops and C-terminal region are involved in the interaction. The appearance of side-chain NH proton resonances of arginine residues of loop 3 and imino proton resonances of RNA bases upon complex formation suggests the formation of intermolecular hydrogen bonds. The structural arrangement of the rings of the conserved aromatic residues of β2 and β3 is suitable for stacking interaction with RNA bases, known to be one of the major protein-RNA interactions, but a survey of the perturbation data suggested that the stacking interaction is not ideally achieved in the complex, which may be related to the weaker RNA-binding of RBD2.

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Ectopic expression of constitutively activated Ral GTPase inhibits cell shape changes during Drosophila eye development

Sawamoto Kazunobu, Yamada Chiharu, Kishida Shosei, Hirota Yuki, Taguchi Akiko, Kikuchi Akira, Okano Hideyuki

Oncogene 18 （ 11 ） 1967 - 1974 1999年03月

ISSN 0950-9232

The small GTP-binding protein Ral is activated by RalGDS, one of the effector molecules for Ras. Active Ral binds to a GTPase activating protein for CDC42 and Rac. Although previous studies suggest a role for Ral in the regulation of CDC42 and Rac, which are involved in arranging the cytoskeleton, its in in vivo function is largely unknown. To examine the effect of overexpressing Ral on development, transgenic Drosophila were generated that overexpress wild-type or mutated Ral during eye development. While wild-type Ral caused no developmental defects, expression of a constitutively activated protein resulted in a rough eye phenotype. Activated Rat did not affect cell fate determination in the larval eye discs but caused severe disruption of the ommatidial organization later in pupal development. Phalloidin staining showed that activated Ral perturbed the cytoskeletal structure and cell shape changes during pupal development. This phenotype is similar to that caused by RhoA overexpression. In addition, the phenotype was synergistically enhanced by the coexpression of RhoA. These results suggest that Ral functions to control the cytoskeletal structure required for cell shape changes during Drosophila development.

Stimulation of myelin basic protein gene transcription by fyn tyrosine kinase for myelination

Umemori Hisashi, Kadowaki Yasunori, Hirosawa Kazushige, Yoshida Yutaka, Hironaka Katsunori, Okano Hideyuki, Yamamoto Tadashi

Journal of Neuroscience 19 （ 4 ） 1393 - 1397 1999年02月

ISSN 0270-6474

Myelin is synthesized about the time of birth. The Src-family tyrosine kinase Fyn is involved in the initial events of myelination. Fyn is present in myelin-forming cells and is activated through stimulation of celt surface receptors such as large myelin-associated glycoprotein (L-MAG). Here we show that Fyn stimulates transcription of the myelin basic protein (MBP) gene for myelination. MBP is a major component of the myelin membrane. In 4-week-old Fyn-deficient mice, MBP is significantly reduced, and electron microscopic analysis showed that myelination is delayed, compared with wild-type mice. The Fyn-deficient mice had thinner, more irregular myelin than the wild- type. We found that Fyn stimulates the promoter activity of the MBP gene by approximately sevenfold. The region responsible for the transactivation by Fyn is located between nucleotides -675 and -647 with respect to the transcription start site. Proteins binding to this region were found by gel shift study, and the binding activity correlates with Fyn activity during myelination. These results suggest that transactivation of the MBP gene by Fyn is important for myelination.

Proapoptotic activity of Caenorhabditis elegans CED-4 protein in Drosophila

Kanuka Hirotaka, Hisahara Shin, Sawamoto Kazunobu, Shoji Shin Ichi, Okano Hideyuki, Miura Masayuki

Proceedings of the National Academy of Sciences of the United States of America 96 （ 1 ） 145 - 150 1999年01月

ISSN 0027-8424

CED-4 protein plays an important role in the induction of programmed cell death in Caenorhabditis elegans through the activation of caspases. However, the precise mechanisms by which it activates caspases remain unknown. To investigate the conservation of CED-4 function in evolution, transgenic Drosophila lines that express CED-4 in the compound eye were generated. Ectopic expression of CED-4 in the eyes induced massive apoptotic cell death through caspase activation. An ATP-binding site (P-loop) mutation in CED-4 (K165R) causes a loss of function in its ability to activate Drosophila caspase, and an ATPase inhibitor blocks the CED-4-dependent caspase activity in Drosophila S2 cells. Immunoprecipitation analysis showed that both CED-4 and CED-4 (K165R) bind directly to Drosophila caspase drICE, and the overexpression of CED-4 (K165R) inhibits CED-4-, ecdysone-, or cycloheximide-dependent caspase activation in S2 cells. Furthermore, CED-4 (K165R) partially prevented cell death induced by CED-4 in Drosophila compound eyes. Thus, CED-4 function is evolutionarily conserved in Drosophila, and the molecular mechanisms by which CED-4 activates caspases might require ATP binding and direct interaction with the caspases.

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A genetic approach to visualization neurotechnique of multisynaptic neural pathways using plant lectin transgene

Yoshihara Yoshihiro, Mizuno Takeo, Nakahira Masakiyo, Kawasaki Miwa, Watanabe Yasuyoshi, Kagamiyama Hiroyuki, Jishage Kou ichi, Ueda Otoya, Suzuki Hiroshi, Tabuchi Katsuhiko, Sawamoto Kazunobu, Okano Hideyuki, Noda Tetsuo, Mori Kensaku

Neuron 22 （ 1 ） 33 - 41 1999年01月

ISSN 0896-6273

The wiring patterns among various types of neurons via specific synaptic connections are the basis of functional logic employed by the brain for information processing. This study introduces a powerful method of analyzing the neuronal connectivity patterns by delivering a tracer selectively to specific types of neurons while simultaneously transsynaptically labeling their target neurons. We developed a novel genetic approach introducing cDNA for a plant lectin, wheat germ agglutinin (WGA), as a transgene under the control of specific promoter elements. Using this method, we demonstrate three examples of visualization of specific transsynaptic neural pathways: the mouse cerebellar efferent pathways, the mouse olfactory pathways, and the Drosophila visual pathways. This strategy should greatly facilitate studies on the anatomical and functional organization of the developing and mature nervous system.

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WRM-1 activates the LIT-1 protein kinase to transduce anterior/posterior polarity signals in C. elegans

Rocheleau C. E., Yasuda J., Tae Ho Shin Ho Shin, Lin R., Sawa H., Okano H., Priess J. R., Davis R. J., Mello C. C.

Cell 97 （ 6 ） 717 - 726 1999年

ISSN 0092-8674

During C. elegans development, Wnt/WG signaling is required for differences in cell fate between sister cells born from anterior/posterior divisions. A β-catenin-related gene, wrm-1, and the lit-1 gene are effectors of this signaling pathway and appear to downregulate the activity of POP-1, a TCF/LEF-related protein, in posterior daughter cells. We show here that lit- 1 encodes a serine/threonine protein kinase homolog related to the Drosophila tissue polarity protein Nemo. We demonstrate that the WRM-1 protein binds to LIT-1 in vivo and that WRM-1 can activate the LIT-1 protein kinase when coexpressed in vertebrate tissue culture cells. This activation leads to phosphorylation of POP-1 and to apparent changes in its subcellular localization. Our findings provide evidence for novel regulatory avenues for an evolutionarily conserved Wnt/WG signaling pathway.

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Involvement of caspases in cytotoxic cytokine-mediated oligodendrocyte cell death

Miura M., Hisahara S., Takano R., Shoji S., Okano H.

Protoplasma 206 （ 4 ） 219 - 223 1999年

ISSN 0033-183X

Oligodendrocytes are myelin-forming cells in the mammalian central nervous system. About 50% of oligodendrocytes undergo cell death in normal development. In addition, massive oligodendrocyte cell death has been observed in multiple sclerosis. Tumor necrosis factor (TNF) is thought to be one of the mediators responsible for the damage of oligodendrocytes in multiple sclerosis. The addition of TNF-α to primary cultures of oligodendrocytes significantly decreased the number of live cells in 72 h. DNA fragmentation was detected in TNF-treated oligodendrocytes at 36h by TUNEL assay. Chemical inhibitors Ac-YVAD-CHO (a specific inhibitor of caspase-1[ICE]-like proteases) as well as Ac-DEVD-CHO (a specific inhibitor of caspase-3[CPP32]-like proteases) enhanced the survival of oligodendrocytes treated with TNF-α, indicating that caspase-1- and the caspase-3-mediated cell-death pathways are activated in TNF-induced oligodendrocyte cell death. Caspase-11 is involved in activation of caspase-1. Oligodendrocytes from CASP-11-deficient mice are partially resistant to TNF-induced oligodendrocyte cell death. Our results suggest that the inhibition of caspases may be a novel approach to treat multiple sclerosis.

Learning and memory

Okano Hideyuki, Hirano Tomoo, Balaban Evan

Proceedings of the National Academy of Sciences of the United States of America 97 （ 23 ） 12403 - 12404 2000年11月

ISSN 0027-8424

Memory is one of the most fundamental mental processes. Neuroscientists study this process by using extremely diverse strategies. Two different approaches aimed at understanding learning and memory were introduced in this symposium. The first focuses on the roles played by synaptic plasticity, especially in long-term depression in the cerebellum in motor learning, and its regulatory mechanism. The second approach uses an elegant chick-quail transplantation system on defined brain regions to study how neural populations interact in development to form behaviorally important neural circuits and to elucidate neurobiological correlates of perceptual and motor predispositions.

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神経細胞の再生

柴田護，岡野栄之

日本老年医学会雑誌 37 （ 10 ） 763-770 2000年10月

研究論文（学術雑誌）, 共著

Caspases determine the vulnerability of oligodendrocytes in the ischemic brain

Shibata Mamoru, Hisahara Shin, Hara Hideaki, Yamawaki Takemori, Fukuuchi Yasuo, Yuan Junying, Okano Hideyuki, Miura Masayuki

Journal of Clinical Investigation 106 （ 5 ） 643-653 2000年09月

研究論文（学術雑誌）, 共著

神経形成における幹細胞と移植

岩本範顕，小川祐人，戸山芳昭，岡野栄之

今日の移植 13 （ 4 ） 343-348 2000年07月

研究論文（学術雑誌）, 共著

Nerve growth factor protects oligodendrocytes from tumor necrosis factor-α-induced injury through Akt-mediated signaling mechanisms

Takano Riya, Hisahara Shin, Namikawa Kazuhiko, Kiyama Hiroshi, Okano Hideyuki, Miurat Masayuki

Journal of Biological Chemistry 275 （ 21 ） 16360 - 16365 2000年05月

ISSN 0021-9258

Tumor necrosis factor-α is thought to be one of the most important inflammatory cytokines associated with the demyelinating disease multiple sclerosis. We determined whether neurotrophins could protect oligodendrocytes from tumor necrosis factor-α-mediated cytotoxicity. Among the neurotrophins tested, nerve growth factor was most effective at preventing cell death. Nerve growth factor also prevented the tumor necrosis factor-induced loss of mitochondrial membrane potential. Overexpression of constitutively active Akt, a downstream target of phosphatidylinositol 3-kinase, but not of constitutively active MEK, protected oligodendrocytes from tumor necrosis factor-induced injury. Moreover, overexpression of dominant-negative Akt negated the protective effects of nerve growth factor on tumor necrosis factor-mediated oligodendrocyte cytotoxicity. These findings indicate that the Akt pathway is crucial in nerve growth factor-mediated oligodendrocyte protection.

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Conditional expression of anti-apoptotic protein p35 by Cre-mediated DNA recombination in cardiomyocytes from loxP-p35-transgenic mice

Araki Takashi, Shibata Mamoru, Takano Riya, Hisahara Shin, Imamura Si, Fukuuchi Yasuo, Saruta Takao, Okano Hideyuki, Miura Masayuki

Cell Death and Differntiation 7 （ 5 ） 485-492 2000年05月

研究論文（学術雑誌）, 共著

Mutations modulating the Argos-regulated signaling pathway in Drosophila eye development

Taguchi Akiko, Sawamoto Kazunobu, Okano Hideyuki

Genetics 154 （ 4 ） 1639 - 1648 2000年04月

ISSN 0016-6731

Argos is a secreted protein that contains an EGF-like domain and acts as an inhibitor of Drosophila EGF receptor activation. To identify genes that function in the Argos-regulated signaling pathway, we performed a genetic screen for enhancers and suppressors of the eye phenotype caused by the overexpression of argos. As a result, new alleles of known genes encoding components of the EGF receptor pathway, such as Star, sprouty, bulge, and clown, were isolated. To study the role of clown in development, we examined the eye and wing phenotypes of the clown mutants in detail. In the eye discs of clown mutants, the pattern of neuronal differentiation was impaired, showing a phenotype similar to those caused by a gain-of-function EGF receptor mutation and overexpression of secreted Spitz, an activating ligand for the EGF receptor. There was also an increased number of pigment cells in the clown eyes. Epistatic analysis placed clown between argos and Ras1. In addition, we found that clown negatively regulated the development of wing veins. These results suggest that the clown gene product is important for the Argos-mediated inhibition of EGF receptor activation during the development of various tissues. In addition to the known genes, we identified six mutations of novel genes. Genetic characterization of these mutants suggested that they have distinct roles in cell differentiation and/or survival regulated by the EGF receptor pathway.

外傷性脊髄損傷に対する神経幹細胞移植の試み

小川祐人，宮田卓樹，澤本和延，渡辺雅彦，戸山芳昭，岡野栄之

日本パラプレジア医学会雑誌 13 （ 1 ） 144-145 2000年04月

研究論文（学術雑誌）, 共著

The interaction between the Drosophila secreted protein Argos and the epidermal growth factor receptor inhibits dimerization of the receptor and binding of secreted spitz to the receptor

Jin Ming Hao, Sawamoto Kazunobu, Ito Mikiko, Okano Hideyuki

Molecular and Cellular Biology 20 （ 6 ） 2098 - 2107 2000年03月

ISSN 0270-7306

Drosophila Argos (Aos), a secreted protein with an epidermal growth factor (EGF)-like domain, has been shown to inhibit the activation of the Drosophila EGF receptor (DER). However, it has not been determined whether Aos binds directly to DER or whether regulation of the DER activation occurs through some other mechanism. Using DER-expressing cells (DER/S2) and a recombinant DER extracellular domain-Fc fusion protein (DER-Fc), we have shown that Aos binds directly to the extracellular domain of DER with its carboxyl-terminal region, including the EGF-like domain. Furthermore, Aos can block the binding of secreted Spitz (sSpi), a transforming growth factor α- like ligand of DER, to the extracellular domain of DER. We observed that sSpi stimulates the dimerization of both the soluble DER extracellular domain (sDER) and the intact DER in the DER/S2 cells and that Aos can block the sSpi-induced dimerization of both sDER and intact DER. Moreover, we have shown that, by directly interacting with DER, Aos and SpiAos (a chimeric protein that is composed of the N-terminal region of Spi and the C-terminal region of Aos) inhibit the dimerization and phosphorylation of DER that are induced by DER's overexpression in the absence of sSpi. These results indicate that Aos exerts its inhibitory function through dual molecular mechanisms: by blocking both the receptor dimerization and the binding of activating ligand to the receptor. This is the first description of this novel inhibitory mechanism for receptor tyrosine kinases.

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In vitro neurogenesis by progenitor cells isolated from the adult human hippocampus

Roy Neeta Singh, Wang Su, Jiang Li, Kang Jian, Benraiss Abdellatif, Harrison-Restelli Catherine, Fraser Richard A R, Couldwell William T., Kawaguchi Ayano, Okano Hideyuki, Nedergaard Maiken, Goldman Steven A.

Nature Medicine 6 （ 3 ） 271 - 277 2000年03月

ISSN 1078-8956

Neurogenesis persists in the adult mammalian hippocampus. To identify and isolate neuronal progenitor cells of the adult human hippocampus, we transfected ventricular zone-free dissociates of surgically-excised dentate gyrus with DNA encoding humanized green fluorescent protein (hGFP), placed under the control of either the nestin enhancer (E/nestin) or the Tα1 tubulin promoter (P/Tα1), two regulatory regions that direct transcription in neural progenitor cells. The resultant P/Tα1:hGFP⁺, and E/nestin:enhanced (E)GFP⁺ cells expressed βIII-tubulin or microtubule- associated protein-2; many incorporated bromodeoxyuridine, indicating their genesis in vitro. Using fluorescence-activated cell sorting, the E/nestin:EGFP⁺ and P/Tα1:hGFP⁺ cells were isolated to near purity, and matured antigenically and physiologically as neurons. Thus, the adult human hippocampus contains mitotically competent neuronal progenitors that can be selectively extracted. The isolation of these cells may provide a cellular substrate for re-populating the damaged or degenerated adult hippocampus.

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Targeted expression of baculovirus p35 caspase inhibitor in oligodendrocytes protects mice against autoimmune-mediated demyelination

Hisahara Shin, Araki Takashi, Sugiyama Fumihiro, Yagami Ken Ichi, Suzuki Misao, Abe Kuniya, Yamamura Ken Ichi, Miyazaki Jun Ichi, Momoi Takashi, Saruta Takao, Bernard Claude C A, Okano Hideyuki, Miura Masayuki

EMBO Journal 19 （ 3 ） 341 - 348 2000年02月

ISSN 0261-4189

The mechanisms underlying oligodendrocyte (OLG) loss and the precise roles played by OLG death in human demyelinating diseases such as multiple sclerosis (MS), and in the rodent model of MS, experimental autoimmune encephalomyelitis (EAE), remain to be elucidated. To clarify the involvement of OLG death in EAE, we have generated transgenic mice that express the baculovirus anti-apoptotic protein p35 in OLGs through the Cre-loxP system. OLGs from cre/p35 transgenic mice were resistant to tumor necrosis factor-α-, anti-Fas antibody- and interferon-γ-induced cell death. cre/p35 transgenic mice were resistant to EAE induction by immunization with the myelin oligodendrocyte glycoprotein. The numbers of infiltrating T cells and macrophages/microglia in the EAE lesions were significantly reduced, as were the numbers of apoptotic OLGs expressing the activated form of caspase-3. Thus, inhibition of apoptosis in OLGs by p35 expression alleviated the severity of the neurological manifestations observed in autoimmune demyelinating diseases.

Promoter-targeted selection and isolation of neural progenitor cells from the adult human ventricular zone

Roy Neeta S., Benraiss Abdellatif, Wang Su, Fraser Richard A R, Goodman Robert, Couldwell William T., Nedergaard Maiken, Kawaguchi Ayano, Okano Hideyuki, Goldman Steven A.

Journal of Neuroscience Research 59 （ 3 ） 321 - 331 2000年02月

ISSN 0360-4012

Adult humans, like their nonhuman mammalian counterparts, harbor persistent neural progenitor cells in the forebrain ventricular lining. In the absence of adequate surface markers, however, these cells have proven difficult to isolate for study. We have previously identified and selected neural progenitor cells from both the fetal and adult rodent ventricular zone (VZ), by sorting forebrain cells transfected with plasmid DNA encoding the gene for green fluorescent protein driven by the early neuronal promoter for Tα1 tubulin (P/Tα1:hGFP). We have now extended this approach by purifying both P/Tα1:hGFP tubulin-defined neuronal progenitors, as well as potentially less committed E/nestin:hGFP-defined neural progenitor cells, from the adult human VZ. The ventricular wall of the temporal horn of the lateral ventricle was dissected from temporal lobes obtained from four adult patients undergoing therapeutic lobectomy. These samples were dissociated, and the cultured cells transduced with either P/Tα1:hGFP or E/nestin:EGFP plasmid DNA. A week later, the cells were redissociated, selected via fluorescence- activated cell sorting (FACS) on the basis of neural promoter-driven GFP expression, and replated. The majority of these cells expressed the early neuronal protein βIII-tubulin upon FACS; within the week thereafter, most matured as morphologically evident neurons that coexpressed βIII-tubulin and microtubule-associated protein (MAP)-2. Many of these neurons had incorporated bromodeoxyuridine in vitro in the days before FACS, indicating their mitogenesis in vitro. Thus, the use of fluorescent transgenes under the control of early neural promoters permits the enrichment of neuronal progenitor cells from the adult human ventricular zone. The specific acquisition, in both purity and number, of residual neural progenitor cells from the adult human brain may now permit hitherto unfeasible studies of both their biology and practical application. (C) 2000 Wiley-Liss, Inc.

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神経幹細胞の分化制御機構と再生医学への応用

赤松和土，岡野栄之

医学のあゆみ 192 （ 8 ） 843-846 2000年02月

研究論文（学術雑誌）, 共著

Drob-1, a Drosophila member of the Bcl-2/CED-9 family that promotes cell death

Igaki Tatsushi, Kanuka Hirotaka, Inohara Naohiro, Sawamoto Kazunobu, Núñez Gabriel, Okano Hideyuki, Miura Masayuki

Proceedings of the National Academy of Sciences of the United States of America 97 （ 2 ） 662 - 667 2000年01月

ISSN 0027-8424

The Bcl-2/CED-9 family of proteins, which includes both antiapoptotic and proapoptotic members, plays key regulating roles in programmed cell death. We report here the identification and characterization of Drob-1, the first Drosophila member of the Bcl-2/CED-9 family to be isolated. Drob-1 contains four conserved Bcl-2 homology domains (BH1, BH2, BH3, and BH4) and a C-terminal hydrophobic domain. Ectopic expression of Drob-1 in the developing Drosophila eye resulted in a rough-eye phenotype. Furthermore, when overexpressed in Drosophila S2 cells, Drob-1 induced apoptosis accompanied by elevated caspase activity. This Drob-1-induced cell death, however, could not be antagonized by baculovirus p35, a broad-spectrum caspase inhibitor. Drob-1 was localized to the intracytoplasmic membranes, predominantly to the mitochondrial membranes, and a mutant Drob-1 lacking the hydrophobic C terminus lost both its mitochondrial localization and its proapoptotic activity. These results suggest that Drob-1 promotes cell death by inducing both caspase-dependent and -independent pathways at the mitochondria. Our identification of Drob-1 and further genetic analysis should provide increased understanding of the universal mechanisms by which the Bcl-2/CED-9 family members and other related proteins regulate apoptosis.

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GAL4/UAS-WGA system as a powerful tool for tracing Drosophila transsynaptic neural pathways

Tabuchi Katsuhiko, Sawamoto Kazunobu, Suzuki Emiko, Ozaki Koichi, Sone Masaki, Hama Chihiro, Tanifuji-Morimoto Takako, Yuasa Yoshihiro, Yoshihara Yoshihiro, Nose Akinao, Okano Hideyuki

Journal of Neuroscience Research 59 （ 1 ） 94 - 99 2000年01月

ISSN 0360-4012

Visualization of specific transsynaptic neural pathways is an indispensable technique for understanding the relationship between structure and function in the nervous system. Here, we demonstrate the application of the wheat germ agglutinin (WGA) transgene technique for tracing transsynaptic neural pathways in Drosophila. The intracellular localization of WGA was examined by immunoelectron microscopy. WGA signals were detected in granule- like structures in both the outer photoreceptor cells expressing WGA and the second-order laminar neurons. Misexpression of tetanus toxin (TNT), which inactivates N-synaptobrevin, in the outer photoreceptor cells resulted in the elimination of on/off transients in electroretinogram (ERG) recordings and in a great reduction in WGA transfer into laminar neurons, suggesting that anterograde WGA transsynaptic transfer is dependent mainly on synaptic transmission. Retrograde WGA transfer was also detected upon its forced expression in muscle cells. WGA primarily expressed in muscle cells was taken up by motoneuron axons and transported to their cell bodies in the ventral nerve cord, suggesting that WGA can trace motoneuronal pathways in combination with the muscle-specific GAL4 driver. Thus, the GAL4/UAS-WGA system should facilitate the dissection of the Drosophila neural circuit formation and/or synaptic activity in various regions and at various developmental stages.

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The bHLH gene Hes1 as a repressor of the neuronal commitment of CNS stem cells

Nakamura Yuki, Sakakibara Shin Ichi, Miyata Takaki, Ogawa Masaharu, Shimazaki Takuya, Weiss Samuel, Kageyama Ryoichiro, Okano Hideyuki

Journal of Neuroscience 20 （ 1 ） 283 - 293 2000年01月

ISSN 0270-6474

Hes1 is one of the basic helix-loop-helix transcription factors that regulate mammalian CNS development, and its loss- and gain-of-function phenotypes indicate that it negatively regulates neuronal differentiation. Here we report that Has1(-/-) mice expressed both early (TuJ1 and Hu) and late (MAP2 and Neurofilament) neuronal markers prematurely, and that there were approximately twice the normal number of neurons in the Hes1 (-/-) brain during early neural development. However, immunochemical analyses of sections and dissociated cells using neural progenitor markers, including nestin, failed to detect any changes in Hes1 (-/-) progenitor population. Therefore, further characterization of neural progenitor cells that discriminated between multipotent and monopotent cells was performed using two culture methods, low-density culture, and a neurosphere assay. We demonstrate that the self-renewal activity of multipotent progenitor cells was reduced in the Hes1 (-/-) brain, and that their subsequent commitment to the neuronal lineage was accelerated. The Hes1 (-/-) neuronal progenitor cells were functionally abnormal, in that they divided, on average, only once, and then generated two neurons, (instead of one progenitor cell and one neuron), whereas wild-type progenitor cells divided more. In addition, some Hes1(-/-) progenitors followed an apoptotic fate. The overproduction of neurons in the early Hes1(-/-) brains may reflect this premature and immediate generation of neurons as well as a net increase in the number of neuronal progenitor cells. Taken together, we conclude that Hes1 is important for maintaining the self- renewing ability of progenitors and for repressing the commitment of multipotent progenitor cells to a neuronal fate, which is critical for the correct number of neurons to be produced and for the establishment of normal neuronal function.

Dissection of signaling cascades through gp130 in vivo

Ohtani Takuya, Ishihara Katsuhiko, Atsumi Toru, Nishida Keigo, Kaneko Yukiko, Miyata Takaki, Itoh Shousaku, Narimatsu Masahiro, Maeda Hisoka, Fukada Toshiyuki, Itoh Motoyuki, Okano Hideyuki, Hibi Masahiko, Hirano Toshio

Immunity 12 （ 1 ） 95 - 105 2000年01月

ISSN 1074-7613

We generated a series of knockin mouse lines, in which the cytokine receptor gp130-dependent STAT3 and/or SHP2 signals were disrupted, by replacing the mouse gp130 gene with human gp130 mutant cDNAs. The SHP2 signal-deficient mice (gp130(F759/F759)) were born normal but displayed splenomegaly and lymphadenopathy and an enhanced acute phase reaction. In contrast, the STAT3 signal-deficient mice (gp130(FXXQ/FXXQ)) died perinatally, like the gp130-deficient mice (gp130(D/D)). The gp13(F759/F759) mice showed prolonged gp130-induced STAT3 activation, indicating a negative regulatory role for SHP2. Th1 -type cytokine production and IgG2a and IgG2b production were increased in the gp13(F759/F759) mice, while they were decreased in the gp130(FXXQ/FXXQ) immune system. These results indicate that the balance of positive and negative signals generated through gp130 regulates the immune responses.

Regulation of neural development mediated by neural RNA binding proteins

Akamatsu W., Okano H.

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 45 （ 3 Suppl ） 227 - 232 2000年

ISSN 0039-9450

Identification and characterization of human brain tumor antigens: a potential target for specific immunotherapy and diagnostic marker

Toda M, Iizuka Y, Takahashi S, Fujita T, Ikeda E, Yoshida K, Kawase T, Okano H, Uyemura K, Kawakami Y

Proceedings of the American Association for Cancer Research 41 710 2000年

研究論文（学術雑誌）, 共著

Musashi 1;An evolutionally conserved marker forCNS progenitor cells including neural stem cells.

Kaneko Yukiko, Imai T, Nakamura Y, Sawamoto K, Ogawa Yuuto, Toyama Yoshiaki, Miyata T, Okano Hideyuki

Developmental Neuroscience 22 139-153 2000年

研究論文（学術雑誌）, 共著

Neural stem cells

Okano H.

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 45 （ 13 Suppl ） 2063 - 2077 2000年

ISSN 0039-9450

Role of caspase-1 subfamily in cytotoxic cytokine-induced oligodendrocyte cell death

Hisahara S., Takano R., Shoji S., Okano H., Miura M.

Journal of Neural Transmission, Supplement （ 58 ） 135 - 142 2000年

ISSN 0303-6995

Oligodendrocytes are myelin forming cells in mammalian central nervous system. About 50% of oligodendrocytes (OLGs) undergo cell death in normal development. In addition, OLG cell deaths have been observed in demyelinating diseases including multiple sclerosis (MS). Clinical observations and in vitro cell culture studies have suggested that cytokines mediate OLG cell damage in multiple sclerosis (MS). Among the cytokines, tumor necrosis factor (TNF) is thought to be one of the mediators responsible for the damage of OLGs in MS. The administration of TNF-α to primary cultures of OLGs induced DNA fragmentation, and significantly decreased the number of live OLGs. Chemical inhibitors Ac-YVAD-CHO (a specific inhibitor of caspase-1 (ICE)-like proteases) enhanced the survival of TNF-α treated OLGs better than Ac-DEVD-CHO (a specific inhibitor of caspase-3 (CPP32)-like proteases). These results indicate that caspase-1-mediated cell-death pathway are activated in TNF-induced OLG cell death. Caspase-11 is involved in activation of caspase-1. Oligodendrocytes from caspase-11-deficient mice are partially resistant to TNF-induced OLG cell death. Our results suggest that the inhibition of caspase-1 sufamily may be a novel therapeutic approach to treat MS.

Neural stem cells

Okano H.

Japanese Journal of Psychopharmacology 20 （ 1 ） 21 - 26 2000年

ISSN 1340-2544

Neural stem cells (NSCs) are multipotential progenitor cells that have self-renewal activities. A single NSC is capable of generating various kinds of cells within the CNS, including neurons, astrocytes, and oligodendrocytes. Because of these characteristics, there is an increasing interest in NSCs and neural progenitor cells from the aspects of both basic developmental biology and therapeutic applications for damaged brain. By understanding the nature of NSCs present in the CNS, extracellular factors and signal transduction cascades involved in the differentiation and maintenance of NSCs, population dynamics and localization of NSCs in embryonic and adult brains, prospective identification and isolation of NSCs, and induction of NSCs into particular neuronal phenotypes, it would be possible to develop a feasible strategy to manipulate cells in situ to treat damaged brain.

Expression patterns of musashi homologs of the ascidians, Halocynthia roretzi and Ciona intestinalis

Kawashima Takeshi, Murakami Akikazu R., Ogasawara Michio, Tanaka Kimio J., Isoda Ryuji, Sasakura Yasunori, Nishikata Takahito, Okano Hideyuki, Makabe Kazuhiro W.

Development Genes and Evolution 210 （ 3 ） 162 - 165 2000年

ISSN 0949-944X

The gene family encoding RNA-binding proteins includes important regulators involved in the neurogenesis in both protostomes and deuterostomes. We isolated cDNAs of the ascidian homolog of one of the RNA-binding proteins, MUSASHI, from Halocynthia roretzi and Ciona intestinalis. The predicted amino acid sequences contained two RNA-recognition and RNA-binding motifs in the N-terminus and an ascidian-specific YG-rich domain in the C-terminus. Maternal transcripts of musashi were ubiquitous in early cleavage-stage embryos. Ascidian musashi had three domains of zygotic expression: the brain, nerve cord, and mesenchyma. The temporal order of the onset in these domains was highly divergent between the two species of ascidian examined.

Identification and selection of neural progenitor cells

Okano H., Goldman S. A.

NeuroScience News 3 （ 6 ） 27 - 31 2000年

ISSN 1027-6599

Neural stem cells (NSCs) are self-renewing and multipotential neural progenitor cells, which have received strong attention as potential tools to treat the injured or diseased brain. To develop therapeutic strategies intended to capitalize upon the plasticity and propagability of NSCs, we need both to better understand their biology, and to develop better means for their prospective identification and harvest. We summerize here recent findings from our laboratories. Specifically, we describe the prospective identification of NSCs, using fluorescent reporters placed under the control of the nestin enhancer. Together, these findings provide us both new conceptual and operational strategies for studying NSCs, and as such should greatly accelerate their development for the treatment of neurologic disease.

MSI-1, a neural RNA-binding protein, is involved in male mating behaviour in Caenorhabditis elegans

Yoda Akinori, Sawa Hitoshi, Okano Hideyuki

Genes to Cells 5 （ 11 ） 885 - 895 2000年

ISSN 1356-9597

Background: Neural RNA-binding proteins are thought to play important roles in neural development and the functional regulation of postmitotic neurones by mediating post-transcriptional gene regulation. RNA-binding proteins belonging to the Musashi family are highly expressed in the nervous system; however, their roles are poorly understood. Results: We identified a Caenorhabditis elegans Musashi homologue, MSI-1, whose RNA-recognition motifs show extensive similarity to those of Drosophila and vertebrate Musashi proteins. We isolated a msi-1 mutant and found males with this mutation to have a mating defect. C. elegans male mating behaviour includes a distinct series of steps: response to contact, backing, turning, vulva location, spicule insertion, and sperm transfer. msi-1 is required for the turning and vulva location steps. Like other Musashi family members, MSI-1 is expressed specifically in neural cells, including male-specific neurones required for turning and vulva location. However, msi-1 was not expressed in proliferating neural progenitors in C. elegans, unlike the Musashi family genes in other systems. Conclusions: Our results suggest that msi-1 is expressed specifically in postmitotic neurones in C. elegans. msi-1 is required for full development of male mating behaviour, possibly through regulation of msi-1 expressing neurones.

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Components of the SWI/SNF complex are required for asymmetric cell division in C. elegans

Sawa Hitoshi, Kouike Hiroko, Okano Hideyuki

Molecular Cell 6 （ 3 ） 617 - 624 2000年

ISSN 1097-2765

Asymmetric cell division is a fundamental process that produces cellular diversity during development. We have identified two mutants in C. elegans (psa-1 and psa-4) in which the asymmetry of T cell division is disrupted. psa-1 and psa-4 encode homologs of yeast SW13 and SW12/SNF2, respectively, which are components of the SWI/SNF complex. We show by RNA interference assay that homologs of other components of SWI/SNF are also involved in T cell division. psa-1 and psa-4 are likely to be required in the T cell during mitosis to cause asymmetric cell division. Because the SWI/SNF complex is required for asymmetric division in S. cerevisiae, these results demonstrate that at least some aspects of the mechanism of asymmetric cell division are conserved between yeast and a multicellular organism.

Neural stem cells

Okano Hideyuki

Clinical Neurology 41 （ 12 ） 1036 - 1040 2001年12月

ISSN 0009-918X

Neural stem cells (NSCs) are multipotential progenitor cells that have self-renewal activities. A single NSC is capable of generating various kinds of cells within the CNS, including neurons, astrocytes, and oligodendrocytes. Because of these characteristics, there is an increasing interest in NSCs and neural progenitor cells from the aspects of both basic developmental biology and therapeutic applications to the damaged brain. By understanding of nature of NSCs present in CNS, extracellular factors and signal transduction cascades involved in the differentiation and maintenance of NSCs, population dynamics and localizations of NSCs in embryonic and adult brains, prospective identification and isolation of NSCs, and induction of NSCs into particular neuronal phenotypes, which will be introduced in this review, it would be possible to develop a feasible strategy to manipulate cells in situ to treat the damaged brain.

In vitro selection法によるRNA結合タンパク質の標的配列の同定

今井貴雄，岡野ジェイムス洋尚，岡野栄之

細胞工学 20 （ 12 ） 1690-1697 2001年12月

研究論文（学術雑誌）, 共著

パーキンソン病治療への新たな布石

澤本和延，岡野栄之

Bio ベンチャー 1 （ 3 ） 90-91 2001年11月

研究論文（学術雑誌）, 共著

Role of Deltex-1 as a transcriptional regulator downstream of the Notch receptor

Yamamoto Naoya, Yamamoto Shin-ichi, Inagaki Fuyuki, Kawaichi Masashi, Fukamizu Akiyoshi, Kishi Noriyuki, Matsuno Kenji, Nakamura Kozo, Weinmaster Gerry, Okano Hideyuki, Nakafuku Masato

Journal of Biological Chemistry 276 （ 48 ） 45031-45040 2001年11月

研究論文（学術雑誌）, 共著

再生治療の海外基礎研究開発状況―特に幹細胞生物学研究を中心に

金村米博，岡野栄之

Science&Technology Jounal 10 （ 00 ） 14-17 2001年11月

研究論文（学術雑誌）, 共著

神経幹細胞の同定・分離の確立と神経再生への挑戦

岡野栄之

神経治療学 18 （ 5/6合併号） 443-449 2001年11月

研究論文（学術雑誌）, 単著

神経幹細胞と再生

今井貴雄，岡野ジェイムス洋尚，岡野栄之

治療学 35 29-35 2001年10月

研究論文（学術雑誌）, 共著

RNA-binding protein Musashi2: Developmentally regulated expression in neural precursor cells and subpopulations of neurons in mammalian CNS

Sakakibara Shin-ich, Nakamura Yuki, Satoh Hitoshi, Okano Hideyuki

Journal of Neuroscince 21 （ 20 ） 8091-8107 2001年10月

研究論文（学術雑誌）, 共著

再生医学の基本戦略：中枢神経系を中心として

岡野栄之

KAST Report 13 （ 1 ） 24-29 2001年10月

研究論文（学術雑誌）, 単著

神経幹細胞：その基礎生物学と高次神経機能・精神神経疾患への応用

岡野栄之

精神神経学雑誌 103 （ 10 ） 759-763 2001年10月

研究論文（学術雑誌）, 単著

幹細胞システムを用いた中枢神経系の再生医学

岡野栄之

Neurologia medico-chirurgica 41 （ extra issue ） 23 2001年10月

研究論文（学術雑誌）, 単著

Brain from bone:Efficient'meta-differentiation'of marrow stroma-derived mature osteoblasts to neurons with Noggin or a demethylating agent

Jun Kohyama, Hitoshi Abe, Takuya Shimazaki, Amane Koizumi, Kin-ichi Nakashima, Satoshi Gojo, Tetsuya Taga, Hideyuki Okano, Jun-ichi Hata, Akihiro Umezawa

Differentiation 68 （ 4-5 ） 235-244 2001年10月

研究論文（学術雑誌）, 共著

IP₃ receptor type 1 mRNAの3'末端非翻訳領域に結合する遺伝子産物と小脳purkinje細胞におけるmRNA局在化機構

飯島崇利，今井貴雄，木村有喜，Bernstein Alan，岡野栄之

神経化学 40 （ 2,3 (Suppl) ） 311 2001年09月

研究論文（学術雑誌）, 共著

An imaging-based 'time-table' of melecular expression in cultured murine neuroepthelial cells

Kawaguchi Ayano, Ogawa Maaharu, Okano Hideyuki, Saito Kaneko, Miyata Takayuki

Neurosciene Research 25 （ Suppl ） S128 2001年09月

研究論文（学術雑誌）, 共著

High-yield selection and extraction of two promoter-defined phenotypes of neural stem cells from the fetal human brain

Keyoung Michael H, Roy Neeta S, Benraiss Abdellatif, Louissaint Abner Jr, Suzuki Akira, Hashimoto Mitsuhiro, Rashbaum William K, Okano Hideyuki, Goldman Steven A

Nature Biotechnology 19 （ 9 ） 843-850 2001年09月

研究論文（学術雑誌）, 共著

Asymmetric inheritance of radial glial fibers by cortical neurons

Miyata Takaki, Kawaguchi Ayano, Okano Hideyuki, Ogawa Masaharu

Neuron 31 （ 5 ） 727-741 2001年09月

研究論文（学術雑誌）, 共著

How does the developing cerebral wall thicken?

Miyata Takaki, Kawaguchi Ayano, Okano Hideyuki, Ogawa Masaharu

Neurosciene Research 25 （ Suppl ） S22 2001年09月

研究論文（学術雑誌）, 共著

Transplantation of FACS-sorted mesencephalic neural progenitor cells and DA neurons into a rat model of Parkinsonism

Okano Hideyuki

Neurosciene Research 25 （ Suppl ） S36 2001年09月

研究論文（学術雑誌）, 単著

中枢神経系の幹細胞生物学

岡野栄之

実験医学　増刊 19 （ 15 ） 2002-2012 2001年09月

研究論文（学術雑誌）, 単著

神経特異的RNA結合蛋白質Huの機能解析

岡野ジェイムス洋尚，赤松和土，矢野真人，早川佳芳，Robert Darnell，岡野栄之

神経化学 40 （ 2,3 (Suppl) ） 367 2001年09月

研究論文（学術雑誌）, 共著

Effect of Mammalian Numb to neural development

Tokunaga Akinori, Nakao Keiko, Okano Hideyuki

Neurosciene Research 25 （ Suppl ） S49 2001年09月

研究論文（学術雑誌）, 共著

Musashi1, an evolutionarily conserved neural RNA-binding protein, is a versatile marker of human glioma cells in determining their cellular origin, malignancy, and proliferative activity

Kanemura Yonehiro, Mori Kanji, Sakakibara Shin-ichi, Fujikawa Hirokazu, Hayashi Hiroshi, Nakano Atsuhisa, Matsumoto Tsuyoshi, Tamura Kazuyoshi, Imai Takao, Ohnishi Takanori, Fushiki Shinji, Nakamura Yasuhiro, Yamasaki Mami, Okano Hideyuki, Arita Norio

Differentiation 68 （ 2-3 ） 141-152 2001年09月

研究論文（学術雑誌）, 共著

The molecular mechanism of mRNA localization in cerebellular purkinje cell

Iijima Takatoshi, Imai Takao, Kimura Yuki, Bernstein Alan, Okano Hideyuki

Neurosciene Research 25 （ Suppl ） S40 2001年09月

研究論文（学術雑誌）, 共著

musashi遺伝子を用いた脳再生治療への道

今井貴雄，岡野栄之

Molecular Medicine 38 （ 8 ） 922-926 2001年08月

研究論文（学術雑誌）, 共著

Self-renewal and isolation of neural stem cells

Okano Hideyuki

Journal of Neurochemisty 78 （ Suppl 1 ） 114 2001年08月

研究論文（学術雑誌）, 単著

Neurogenesis by progenitor cells in the ischemic adult rat hippocampus

Yagita Yoshiki, Kitagawa Kazuo, Otsuki Toshio, Takasawa Ken-ichiro, Miyata Takaki, Okano Hideyuki, Hori Masatsugu, Matsumoto Masayasu

Stroke 32 （ 8 ） 1890-1896 2001年08月

研究論文（学術雑誌）, 共著

Direct isolation of committed neuronal progenitor cells from transgenic mice coexpressing spectrally distinct fluorescent proteins regulated by stage-specific neural promoters

Sawamoto Kazunobu, Yamamoto Atsuyo, Kawaguchi Ayano, Yamaguchi Masahiro, Mori Kensaku, Goldman Steven A, Okano Hideyuki

Journal of Neuroscience Research 65 （ 3 ） 220-227 2001年08月

研究論文（学術雑誌）, 共著

Genetic and functional analysis of PARP, a DNA strand break-binding enzyme

Uchida Masahiro, Hanai Shuji, Uematsu Naoya, Sawamoto Kazunobu, Okano Hideyuki, Miwa Masanao, Uchida Kazuhiko

Mutation Research 477 （ 1-2 ） 89-96 2001年07月

研究論文（学術雑誌）, 共著

神経幹細胞―脳の発生と神経の再生―

吉崎崇仁，岡野栄之

遺伝別冊（ 13 ） 72-80 2001年07月

研究論文（学術雑誌）, 共著

The neural RNA-binding protein Musashi1 translationally regulaties mammalian numb gene expression by interacting with its mRNA

Imai Takao, Tokunaga Akinori, Yoshida Tetsu, Hashimoto Mitsuhiro, Mikoshiba Katsuhiko, Gerry Weinmaster, Nakafuku Masato, Okano Hideyuki

Molecular and Cellular Biology 21 （ 12 ） 3888-3900 2001年06月

研究論文（学術雑誌）, 共著

Activation of murine cytomegalovirus immediate-early promoter in cerebral ventricular zone and glial progenitor cells in transgenic mice

Li Ren-Yong, Baba Satoshi, Kosugi Isao, Arai Yoshifumi, Kawasaki Hideya, Shinmura Yuichiro, Sakakibara Shin-Ichi, Okano Hideyuki, Tsutsui Yoshihiro

Glia 35 （ 1 ） 41-52 2001年06月

研究論文（学術雑誌）, 共著

発達脳における神経細胞の移動：新しいニューロン移動法とその原理

宮田卓樹，川口綾乃，岡野栄之，小川正晴

生体の科学 52 （ 3 ） 224-229 2001年06月

研究論文（学術雑誌）, 共著

神経幹細胞

村山綾子，岡野栄之

bios 6 （ 3 ） 7-8 2001年06月

研究論文（学術雑誌）, 共著

Generation of dopaminergic neurons in the adult brain from mesencephalic precursor cells labeled with a nestin-GFP transgene

Sawamoto Kazunobu, Nakao Naoyuki, Kakishita Koji, Ogawa Yuto, Toyama Yoshiaki, Yamamoto Atsuyo, Yamaguchi Masahiro, Mori Kensaku, Steven A Goldman, Itakura Toru, Okano Hideyuki

Journal of Neurosciense 21 （ 11 ） 3895-3903 2001年06月

研究論文（学術雑誌）, 共著

Translational repression determines a neuronal potential in Drosophila asymmetric cell division

Okabe Masataka, Imai Takao, Kurusu Mitsuhiko, Hiromi Yasushi, Okano Hideyuki

Nature 411 （ 6833 ） 94-98 2001年05月

研究論文（学術雑誌）, 共著

The hiiragi gene encodes a poly(A) polymerase, which controls the formation of the wing margin in Drosophila melanogaster

Murata Takehide, Nagaso Hideyuki, Kashiwabara Shin-ichi, Baba Tadashi, Okano Hideyuki, Yokoyama K Kazunari

Developmental Biology 233 （ 1 ） 137-147 2001年05月

研究論文（学術雑誌）, 共著

Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons

Sawamoto Kazunobu, Nakao Naoyuki, Kobayashi Kazuto, Matsushita Natsuki, Takahashi Hiroshi, Kakishita Koji, Yamamoto Atsuyo, Yoshizaki Takahito, Terashima Toshio, Murakami Fujio, Itakura Toru, Okano Hideyuki

Proceedings of National Academy of Sciences of the United States of America 98 （ 11 ） 6423-6428 2001年05月

研究論文（学術雑誌）, 共著

Expression of mouse igf2 mRNA-binding protein 3 and its implications for the developing central nervous system

Mori Hiroshi, Sakakibara Shin-ichi, Imai Takao, Nakamura Yuki, Iijima Takatoshi, Suzuki Akira, Yuasa Yoshihiro, Takeda Masatoshi, Okano Hideyuki

Journal of Neuroscience Research 64 （ 2 ） 132-143 2001年04月

研究論文（学術雑誌）, 共著

Expression of the neural RNA-binding protein Musashi1 in human gliomas

Toda M, Iizuka Y, Yu W, Imai T, Ikeda E, Yoshida K, Kawase T, Kawakami Y, Okano H, Uyemura K

Glia 34 （ 1 ） 1-7 2001年04月

研究論文（学術雑誌）, 共著

ES細胞を用いた再生医療への戦略

岸憲幸，岡野栄之

産婦人科の世界 53 （増刊号） 264-269 2001年04月

研究論文（学術雑誌）, 共著

神経幹細胞と臨床応用

森裕，武田雅俊，岡野栄之

Dementia Japan 15 （ 1 ） 68-74 2001年04月

研究論文（学術雑誌）, 共著

Murine homologs of deltex define a novel gene family involved in vertebrate Notch signaling and neurogenesis

Kishi Noriyuki, Tang Zhenyu, Maeda Yusuke, Hirai Asami, Mo Rong, Ito Mikiko, Suzuki Satoshi, Nakao Keiko, Kinoshita Tsutomu, Kadesch Tom, Hui Chi Chung, Artavanis-Tsakonas Spyros, Okano Hideyuki, Matsuno Kenji

International Journal of Developmental Neuroscience 19 （ 1 ） 21 - 35 2001年02月

ISSN 0736-5748

Notch signaling plays an important role in cell-fate specification in multicellular organisms by regulating cell-cell communication. The Drosophila deltex gene encodes a modulator of the Notch pathway that has been shown to interact physically with the Ankyrin repeats of Notch. We isolated four distinct cDNAs corresponding to mouse homologs of deltex - mouse Deltex1 (MDTX1), mouse Deltex2 (MDTX2), mouse Deltex2ΔE (MDTX2ΔE), and mouse Deltex3 (MDTX3). Deduced amino acid sequences of these four cDNAs showed a high degree of similarity to Drosophila Deltex and its human homolog, DTX1 throughout their lengths, even though they possess distinct structural features. MDTX proteins formed homotypic and heterotypic multimers. We found that these genes were expressed in the central, peripheral nervous system and in the thymus, overlapping with those of mouse Notch1. In mammalian tissue culture cells, overexpression of any of the four mouse deltex homologs suppressed the transcriptional activity of E47, a basic helix-loop-helix (bHLH) protein, in a manner similar to suppression by an activated form of human Notch1 or human DTX1. In addition, overexpression of MDTX2 and MDTX2ΔE in C2C12 cells under differentiation-inducing conditions suppressed the expression of myogenin, one of the myogenic transcriptional factors; this was also similar to a previously reported activity of constitutively activated Notch. Furthermore, misexpression of any of the MDTX genes in Xenopus embryos resulted in an expansion of the region expressing the neural cell adhesion molecule (N-CAM) gene, a marker for the neuroepithelium. Collectively, our results suggest that these mouse deltex homologs are involved in vertebrate Notch signaling and regulation of neurogenesis. Copyright © 2001 Elsevier Science Ltd.

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Caspase-11 mediates oligodendrocyte cell death and pathogenesis of autoimmune-mediated demyelination

Hisahara S., Yuan J., Momoi T., Okano H., Miura M.

Journal of Experimental Medicine 193 （ 1 ） 111 - 122 2001年01月

ISSN 0022-1007

Multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), are inflammatory diseases of the central nervous system (CNS) characterized by localized areas of demyelination. The mechanisms underlying oligodendrocyte (OLG) injury in MS and EAE remain unknown. Here we show that caspase-11 plays crucial roles in OLG death and pathogenesis in EAE. Caspase-11 and activated caspase-3 were both expressed in OLGs in spinal cord EAE lesions. OLGs from caspase-11-deficient mice were highly resistant to the cell death induced by cytotoxic cytokines. EAE susceptibility and cytokine concentrations in the CNS were significantly reduced in caspase-11-deficient mice. Our findings suggest that OLG death is mediated by a pathway that involves caspases-11 and -3 and leads to the demyelination observed in EAE.

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Nestin-EGFP transgenic mice

Kawaguchi Ayano, Miyata Takaki, Sawamoto Kazunobu, Takashita Noriko, Murayama Ayako, Akamatsu Wado, Ogawa Masaharu, Okabe Masaru, Tano Yasuo, Goldman Steven A., Okano Hideyuki

Molecular and Cellular Neuroscience 17 （ 2 ） 259 - 273 2001年

ISSN 1044-7431

We generated transgenic mice carrying enhanced green fluorescent protein (EGFP) under the control of the nestin second-intronic enhancer (E/nestin:EGFP). Flow cytometry followed by in vitro assays revealed that in situ EGFP expression in the embryonic brain correlated with the mitotic index, the cogeneration of both neurons and gila, and the frequency of neurosphere formation in vitro. High-level EGFP expressors derived from embryos included a distinct subpopulation of cells that were self-renewable and multipotent, criteria that define neural stern cells (NSCs). Such cells were largely absent among lower-level or non-EGFP expressors, thereby permitting us to enrich for NSCs using EGFP expression level. In adults, although E/nestin:EGFP-positive cells included the NSC population, the frequency of neurosphere formation did not correlate directly with the level of EGFP expression. However, moderately EGFP-expressing cells in adults gained EGFP intensity when they formed neurospheres, suggesting embryonic and adult NSCs exist in different microenvironments in vivo.

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Neural stem cell, as a source of graft material for transplantation in neuronal disease

Akamatsu W., Okano H.

No To Hattatsu 33 （ 2 ） 114 - 120 2001年

ISSN 0029-0831

Self-renewing and multipotent neural stem cells are present in the adult human brain. We successfully harvested neural stem cells from mice and humans using misexpressed EGFP proteins under the control of the nestin second intron enhancer. High-level EGFP expressors derived from mouse embryos included a distinct subpopulation of cells that were self-renewable and multipotent. Further, we obtained that neural progenitor cells from rat fetal spinal cords using a neurosphere technique, and demonstrated their ability to divide and differentiate into neurons in vivo, where they were integrated into the host tissue in the injured rat spinal cord with resultant behavioral improvement of the recipient rat. We also harvested tyrosine hydroxylase-positive neurons from a transgenic mouse expressing GFP under the control of the tyrosine hydroxylase promoter, and successfully transplanted them into the striatum of rats with parkinsonism with marked improvement of the neurological symptoms. Since neural stem cells can adapt well in the host CNS, studies should focus on their application as a vector in gene therapy and on the introduction in vivo or ex vivo of genes to control their proliferation and differentiation. Neural stem cells are a potential, useful source for developing new therapy for CNS disorders.

Critical role of Caenorhabditis elegans homologs of Cds1 (Chk2)-related kinases in meiotic recombination

Oishi I., Iwai K., Kagohashi Y., Fujimoto H., Kariya K. I., Kataoka T., Sawa H., Okano H., Otani H., Yamamura H., Minami Y.

Molecular and Cellular Biology 21 （ 4 ） 1329 - 1335 2001年

ISSN 0270-7306

Although chromosomal segregation at meiosis I is the critical process for genetic reassortment and inheritance, little is known about molecules involved in this process in metazoa. Here we show by utilizing double-stranded RNA (dsRNA)-mediated genetic interference that novel protein kinases (Ce-CDS-1 and Ce-CDS-2) related to Cds1 (Chk2) play an essential role in meiotic recombination in Caenorhabditis elegans. Injection of dsRNA into adult animals resulted in the inhibition of meiotic crossing over and induced the loss of chiasmata at diakenesis in oocytes of F₁ animals. However, electron microscopic analysis revealed that synaptonemal complex formation in pachytene nuclei of the same progeny of injected animals appeared to be normal. Thus, Ce-CDS-1 and Ce-CDS-2 are the first example of Cds1-related kinases that are required for meiotic recombination in multicellular organisms.

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tincar encodes a novel transmembrane protein expressed in the Tinman-expressing cardioblasts of Drosophila

岡野栄之

MECHANISMS OF DEVELOPMENT 119 S279 - S283 2002年12月

研究論文（学術雑誌）, ISSN 0925-4773

中枢神経系の再生医療

金子慎二郎，岩波明生，中村雅也，小川祐人，戸山芳昭，岡野栄之

日本小児科学会雑誌 106 （ 12 ） 1736-1743 2002年12月

研究論文（学術雑誌）, 共著

教育研修講座―神経幹細胞と損傷脊髄の再生

岩波明生，中村雅也，小川祐人，金子慎二郎，戸山芳昭，岡野栄之

日本整形外科学会誌 76 （ 12 ） 579-591 2002年12月

研究論文（学術雑誌）, 共著

小児科領域における最先端医療＜再生医療の最近の進歩＞―中枢神経系の再生医療

金子慎二郎，岩波明生，中村雅也，小川祐人，戸山芳昭，岡野栄之

日本小児科学会雑誌 106 （ 12 ） 1736-1743 2002年12月

研究論文（学術雑誌）, 共著

神経幹細胞の分化制御

岡田誠司，岡野栄之

Brain medical 14 （ 4 ） 14-21 2002年12月

研究論文（学術雑誌）, 共著

光る脳みそ

村山綾子，岡野栄之

細胞工学 21 （ 12 ） 1405-1405 2002年12月

研究論文（学術雑誌）, 共著

ヒト神経幹細胞移植

島崎琢也，岡野栄之

神経研究の進歩 46 （ 6 ） 790-797 2002年12月

研究論文（学術雑誌）, 共著

tincar encodes a novel transmembrane protein expressed in the Tinman-expressing cardioblasts of Drosophila

Hirota Yuki, Sawamoto Kazunobu, Okano Hideyuki

Gene expression patterns 2 （ 3-4 ） 323-327 2002年12月

研究論文（学術雑誌）, 共著

神経幹細胞と損傷脊髄の再生

岩波明生，小川祐人，中村雅也，金子慎二郎，戸山芳昭，岡野栄之

日本整形外科学会雑誌 76 （ 12 ） 579-589 2002年12月

研究論文（学術雑誌）, 共著

RNA-binding protein Musashi family

Sakakibara Shin ichi, Nakamura Yuki, Yoshida Tetsu, Shibata Shinsuke, Koike Masato, Takano Hiroshi, Ueda Shuichi, Uchiyama Yasuo, Noda Tetsuo, Okano Hideyuki

Proceedings of the National Academy of Sciences of the United States of America 99 （ 23 ） 15194 - 15199 2002年11月

ISSN 0027-8424

Homologues of the Musashi family of RNA-binding proteins are evolutionarily conserved across species. In mammals, two members of this family, Musashil (Msil) and Musashi2 (Msi2), are strongly coexpressed in neural precursor cells, including CNS stem cells. To address the in vivo roles of msi in neural development, we generated mice with a targeted disruption of the gene encoding Msil. Homozygous newborn mice frequently developed obstructive hydrocephalus with aberrant proliferation of ependymal cells in a restricted area surrounding the Sylvius aqueduct. These observations indicate a vital role for msil in the normal development of this subpopulation of ependymal cells, which has been speculated to be a source of postnatal CNS stem cells. On the other hand, histological examination and an in vitro neurosphere assay showed that neither the embryonic CNS development nor the self-renewal activity of CNS stem cells in embryonic forebrains appeared to be affected by the disruption of msil, but the diversity of the cell types produced by the stem cells was moderately reduced by the msil deficiency. Therefore, we performed antisense ablation experiments to target both msi1 and msi2 in embryonic neural precursor cells. Administration of the antisense peptidenucleotides, which were designed to specifically down-regulate msi2 expression, to msi1^-/- CNS stem cell cultures drastically suppressed the formation of neurospheres in a dose-dependent manner. Antisense-treated msi1^-/- CNS stem cells showed a reduced proliferative activity. These data suggest that msi1 and msi2 are cooperatively involved in the proliferation and maintenance of CNS stem cell populations.

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An approach for Parkinson's disease. ALS, and spinal cord injury

Okano Hideyuki

Clinical Neurology 42 （ 11 ） 1143 2002年11月

ISSN 0009-918X

The 43rd Annual Meeting of the Japanese Society of Neurology. Symposium V

Sobue Gen, Okano Hideyuki

Clinical Neurology 42 （ 11 ） 1141 2002年11月

ISSN 0009-918X

RNA-binding protein Musashi family: roles for CNS stem cells and a subpopulation of ependymal cells revealed by targeted disruption and antisense ablation

Sakakibara Shin-ichi, Nakamura Yuki, Yoshida Tetsu, Shibata Shinsuke, Koike Masato, Takano Hiroshi, Ueda Shuichi, Uchiyama Yasuo, Noda Tetsuo, Okano Hideyuki

Proceedings of the National Academic Science of the United States of America 99 （ 23 ） 15194-15199 2002年11月

研究論文（学術雑誌）, 共著

Isolation and transplantation of dopaminergic neurons and neural stem cells

Okano Hideyuki, Yoshizaki Takahito, Shimazaki Takuya, Sawamoto Kazunobu

Parkinsonism and related disorders 9 （ 1 ） 23-28 2002年10月

研究論文（学術雑誌）, 共著

Transplantation of in vitro-expanded fetal neural progenitor cells results in neurogenisis and fuctional recovery after spinal cord contusion injury in adult rats

Ogawa Yuto, Sawamoto Kazunobu, T Miyata, S Miyano, Watanabe Masahiko, Nakamura Masaya, Brebman BS, M Koike, Uchiyama Y, Toyama Yoshiaki, Okano H

Journal of neuroscience research 69 （ 6 ） 925-933 2002年09月

研究論文（学術雑誌）, 共著

神経幹細胞と中枢神経再生

戸田正博，岡野栄之

炎症・再生 22 （ 5 ） 469-473 2002年09月

研究論文（学術雑誌）, 共著

Differential expression of Musashi1 and nestin in the adult rat hippocampus after ischemia

Yagita Yoshiki, Kitagawa Kazuo, Sasaki Tsutomu, Miyata Takaki, Okano Hideyuki, Hori Masatsugu, Matsumoto Masayasu

Jounal of neuroscience research 69 （ 6 ） 750-756 2002年09月

研究論文（学術雑誌）, 共著

Nestin enhancer requirements for expression in normal and injured adult CNS

Johansson Clas B, Lothian Carina, Molin Magnus, Okano Hideyuki, Lendahl Urban

Jounal of neuroscience research 69 （ 6 ） 784-794 2002年09月

研究論文（学術雑誌）, 共著

Evaluation of in vitro proliferative activity of human fetal neural stem/progenitor cells using indirect measurements of viable cells based on cellular metabolic activity

Kanemura Yonehiro, Mori Hideki, Kobayashi Satoshi, Islam Omedul, Kodama Eri, Yamamoto Atsuyo, Nakanishi Youko, Arita Norio, Yamasaki Mami, Okano Hideyuki, Hara Masayuki, Miyake Jun

Jounal of neuroscience research 69 （ 6 ） 869-879 2002年09月

研究論文（学術雑誌）, 共著

Stem cell biology of the central nervous system

Okano Hideyuki

Jounel of neuroscience research 69 （ 6 ） 698-707 2002年09月

研究論文（学術雑誌）, 単著

Neural stem cells: progression of basic research and perspective for clinical application

Okano Hideyuki

Keio jounel of medicine 51 （ 3 ） 115-128 2002年09月

研究論文（学術雑誌）, 単著

Flow cytometric analysis of neural stem cells in the developing and adult mouse brain

Murayama Ayako, Matsuzaki Yumi, Kawaguchi Ayano, Shimazaki Takuya, Okano Hideyuki

Jounal of neuroscience research 69 （ 6 ） 837-847 2002年09月

研究論文（学術雑誌）, 共著

Reaper-mediated inhibition of DIAP1-induced DTRAF1 degradation results in activation of JNK in Drosophila

Kuranaga Erina, Kanuka Hirotaka, Igaki Tatsushi, Sawamoto Kazunobu, Ichijo H, Okano Hideyuki, Miura Masayuki

Nature cell biology 4 （ 9 ） 705-710 2002年09月

研究論文（学術雑誌）, 共著

アプローチ：中枢神経再生への挑戦

岡田洋平，岡野栄之

最新医学 57 （ 7 ） 1583-1590 2002年07月

研究論文（学術雑誌）, 共著

中枢神経系再生への戦略―序文にかえて―

岡野栄之

最新医学 57 （ 7 ） 1581-1582 2002年07月

研究論文（学術雑誌）, 単著

Regulation of tau RNA maturation by thyroid hormone is mediated by the neural RNA-binding protein Musashi-1

Cuadrado Ana, Garcia-Fernandez Luis F, Imai Takao, Okano Hideyuki, Munoz Alberto

Molecular and cellular neuroscience 20 （ 2 ） 198-210 2002年06月

研究論文（学術雑誌）, 共著

損傷脊髄への神経幹細胞移植

中村雅也，戸山芳昭，岡野栄之

実験医学 20 （ 9 ） 1301-1306 2002年06月

研究論文（学術雑誌）, 共著

神経幹細胞の最近の進歩と知見

金城謙太郎，池田康夫，岡野栄之

血液・腫瘍科 44 （ 6 ） 466-470 2002年06月

研究論文（学術雑誌）, 共著

神経細胞の再生医学

小沢洋子，岡野栄之

ゲノム医学 2 （ 3 ） 79-84 2002年06月

研究論文（学術雑誌）, 共著

rugose (rg), a Drosophila a kinase anchor protein, is required for retinal pattern formation and interacts genetically with multiple signaling pathways

Shamloula Hoda K, Mbogho Mkajuma P, Pimentel Angel C, Chrzanowska-Lightowlers Zosia M A, Hyatt Vanneta, Okano Hideyuki, Venkatech, Tadmiri R

Genetics 161 （ 2 ） 693-710 2002年06月

研究論文（学術雑誌）, 共著

概論　中枢神経系の再生戦略

岡野栄之

実験医学 20 （ 9 ） 1276-1279 2002年06月

研究論文（学術雑誌）, 単著

パーキンソン病への細胞移植：モデル動物

吉崎崇仁，澤本和延，岡野栄之

実験医学 20 （ 9 ） 1285-1290 2002年06月

研究論文（学術雑誌）, 共著

Upregulation of Akt phosphorylation at the early stage of middle cerebral artery occlusion in mice

Shibata Mamoru, Yamawaki Takemori, Sasaki Takahiro, Hattori Hidenori, Hamada Junichi, Fukuuchi Yasuo, Okano Hideyuki, Miura Masayuki

Brain research 942 （ 1-2 ） 1-10 2002年06月

研究論文（学術雑誌）, 共著

神経幹細胞を用いたparkinson病治療の可能性

澤本和延，岡野栄之

医学のあゆみ 201 （ 5 ） 325-329 2002年05月

研究論文（学術雑誌）, 共著

神経幹細胞 ―What's new on neural stem cells and other stem cells?―

岡野栄之

医学のあゆみ 201 （ 5 ） 285-289 2002年05月

研究論文（学術雑誌）, 単著

Musashi

Okano Hideyuki, Imai Takao, Okabe Masataka

Journal of Cell Science 115 （ 7 ） 1355 - 1359 2002年04月

ISSN 0021-9533

Transcription is thought to have a major role in the regulation of cell fate; the importance of translational regulation in this process has been less certain. Recent findings demonstrate that translational regulation contributes to cell-fate specification. The evolutionarily conserved, neural RNA-binding protein Musashi, for example, controls neural cell fate. The protein functions in maintenance of the stem-cell state, differentiation, and tumorigenesis by repressing translation of particular mRNAs. In mammals it might play an important role in activating Notch signalling by repressing translation of the Notch inhibitor m-Numb.

神経分化を制御する神経系特異的RNA結合蛋白質

今井貴雄，岡部正隆，赤松和土，岡野ジェイムス洋尚，岡野栄之

生体の科学 53 （ 2 ） 84-90 2002年04月

研究論文（学術雑誌）, 共著

神経栄養因子による脊髄由来神経幹細胞の分化制御―特に内因性神経栄養因子に着目して

中村雅也，岡野栄之，戸山芳昭

臨床整形外科 37 （ 4 ） 475-480 2002年04月

研究論文（学術雑誌）, 共著

神経栄養因子による脊髄由来神経幹細胞の分化制御

中村雅也，岡野栄之，戸山芳昭

臨床整形外科 37 475-480 2002年04月

研究論文（学術雑誌）, 共著

Elevated level of basic fibroblast growth factor in multiple myeloma correlates with increased disease activity

Sato N, Hattori Y, Du W, Yamada Taketo, Kamata T, Kakimoto T, Okamoto S, Kawamura C, Kizaki M, Shimada N, Ote Y, Hata Junichi, Ikeda Yasuo

Japanese journal of cancer research 93 459-466 2002年04月

研究論文（学術雑誌）, 共著

Expression of the Musashi1 gene encoding the RNA-binding protein in human hepatoma cell lines

Shu Hong-Jin, Saito Takafumi, Watanabe Hisayoshi, Ito Jun-Itu, Takeda Hiroaki, Okano Hideyuki, Kawata Sumio

Biochemical and Biophysical Research Communications 293 （ 1 ） 150-154 2002年04月

研究論文（学術雑誌）, 共著

Musashi: a translational regulator of cell fate

Okano Hideyuki, Imai Takao, Okabe Masataka

Jurnal of cell science 115 （ 7 ） 1355-1359 2002年04月

研究論文（学術雑誌）, 共著

ドーパミンニューロン及び前駆細胞の可視化・分離と移

澤本和延，岡野栄之

神経研究の進歩 46 （ 2 ） 247-255 2002年04月

研究論文（学術雑誌）, 共著

神経幹細胞と損傷脊髄の再生

岡野栄之

The Journal of the Japanese Orthopaedic Association 76 （ 3 (Suppl) ） S323 2002年03月

研究論文（学術雑誌）, 単著

新生ラット脊髄損傷に対する神経幹細胞移植

中村雅也，岡野栄之，小川祐人，岩波明生，金子慎二郎，Bergman Barbara S

Acta Anatomica Nipponica 77 （ Suppl ） 9 2002年03月

研究論文（学術雑誌）, 共著

損傷脊髄内サイトカイン・神経栄養因子mRNAの発現の変化

中村雅也，岡野栄之，千葉一裕，Bregman Barbara，戸山芳昭

日本パラプレジア医学会雑誌 15 （ 1 ） 126-127 2002年03月

研究論文（学術雑誌）, 共著

Increased proliferation of neural progenitor cells but reduced survival of newborn cells in the contralateral hippocampus after focal cerebral ischemia in rats

Takasawa Ken-ichiro, Kitagawa Kazuo, Yagita Yoshiki, Sasaki Tsutomu, Tanaka Shigeru, Matsushita Kohji, Ohstuki Toshio, Miyata Takaki, Okano Hideyuki, Hori Masatsugu, Matsumoto Masayasu

Journal of Cerebral Blood Flow and Metabolism 22 （ 3 ） 299-307 2002年03月

研究論文（学術雑誌）, 共著

Overexpression of poly(ADP-ribose) polymerase disrupts organization of cytoskeletal F-actin and tissue polarity in Drosophila

Uchida Masahiro, Hanai Shuji, Uematsu Naoya, Sawamoto Kazuhiro, Okano Hideyuki, Miwa Masanao, Uchida Kazuhiko

Journal of Biological Chemistry 277 （ 8 ） 6696-6702 2002年02月

研究論文（学術雑誌）, 共著

DN-Cadherin is required for spatial arrangement of nerve terminals and ultrastructural organization of synapses

Iwai Youichi, Hirota Yuki, Ozaki Koichi, Okano Hideyuki, Takeichi Masatoshi, Uemura Tadashi

Molecular and Cellular Neurosciences 19 （ 3 ） 375-388 2002年02月

研究論文（学術雑誌）, 共著

Involvement of a proline-rich motif and RING-H2 finger of Deltex in the regulation of Notch signaling

Matsuno Kenji, Ito Mikiko, Hori Kazuya, Miyashita Fumiyasu, Suzuki Satoshi, Kishi Noriyuki, Artavanis-Tsakonas Spyros, Okano Hideyuki

Development 129 （ 4 ） 1049-1059 2002年02月

研究論文（学術雑誌）, 共著

RNA binding protein Musashi1 is expressed in sertoli cells in the rat testis from fetal life to adulthood

Saunders PTK, Maguire SM, Macpherson S, Fenelon MC, Sakakibara Shin-ichi, Okano Hideyuki

Biology of Reproduction 66 （ 6 ） 500-507 2002年02月

研究論文（学術雑誌）, 共著

神経幹細胞とその医療応用について

金村米博，原正之，岡野栄之

生体材料 20 （ 2 ） 39-45 2002年02月

研究論文（学術雑誌）, 共著

幹細胞システムを用いた神経再生の試み

岡野栄之

日本臨床分子医学会記録 38 25 2002年02月

研究論文（学術雑誌）, 単著

神経幹細胞

芝田晋介，岡野栄之

分子細胞治療学 1 （ 1 ） 132-133 2002年02月

研究論文（学術雑誌）, 共著

神経幹細胞：その同定・分離法の開発と移植による神経機能回復の試み

岡野栄之

日本老年医学会雑誌 39 （ 1 ） 44-47 2002年01月

研究論文（学術雑誌）, 単著

神経幹細胞の同定法とその応用

金村米博，岡野栄之

CLINICAL NEUROSCIENCE 別冊 20 （ 1 ） 14-19 2002年01月

研究論文（学術雑誌）, 共著

神経幹細胞の同定・分離法の確立と神経再生への挑戦

岡野栄之

生化学 74 （ 1 ） 17-26 2002年01月

研究論文（学術雑誌）, 単著

Identification of Musashi1-positive cells in human normal and neoplastic neuroepithelial tissues by immunohistochemical methods.

Kanemura Yonehiro, Sakakibara Shin ichi, Okano Hideyuki

Methods in molecular biology (Clifton, N.J.) 198 273 - 281 2002年

ISSN 1064-3745

特集―中枢神経系の再生に挑む；損傷脊髄への神経幹細胞移植

中村雅也，戸山芳昭，岡野栄之

実験医学 20 （ 9 ） 1301-1306 2002年

研究論文（学術雑誌）, 共著

損傷脊髄内サイトカイン・神経栄養因子mRNAの発現の変化

中村雅也，岡野栄之，千葉一裕，Bregman BS，戸山芳昭

日本パラプレジア医学会誌 15 （ 1 ） 126-127 2002年

研究論文（学術雑誌）, 共著

神経疾患における再生医療

'松本有史,　岡野栄之'

日本内科学会雑誌 92 （ 12 ） 110-116 2003年12月

研究論文（学術雑誌）, 共著

幹細胞システムを用いた中枢神経の再生医学

'飯島崇利,　岡野栄之'

神経研究の進歩 47 （ 6 ） 841-849 2003年12月

研究論文（学術雑誌）, 単著

Growth and differentiation potential of main- and side-population cells derived from murine skeletal muscle.

'Tamaki Tsturo, Akatsuka Akira, Okada Yoshinori, Matsuzaki Yumi, Okano Hideyuki, Kimura Minoru'

Experimental Cell Research 291 （ 1 ） 83 - 90 2003年11月

研究論文（学術雑誌）, 共著, 査読有り

Regeneration of central nervous system

Okano Hideyuki, Yoshizaki Takahito, Okada Seiji

Clinical Neurology 43 （ 11 ） 824 - 826 2003年11月

ISSN 0009-918X

It had been long believed that our adult mammalian central nervous system (CNS) does not regenerate after damage due to injuries or degenerative diseases, as Santiago Ramóny Cajal had indicated long time ago. Today, however, CNS came to be recognized as an important target of so called "regenerative medicine". We have been proposing that regeneration of CNS does include the following three concepts: i) re-growth of the damaged neuronal axons, ii) replenishment of lost neural (or neuronal) cells and iii) recovery of lost neural functions. Here, we would like to emphasize that to recapitulate normal neural development is an essential strategy for CNS-regeneration. In this review, we would like to take Parkinson's disease and spinal cord injury as examples to discuss actual strategy aiming for CNS-regeneration.

神経幹細胞の治療への応用

'安達一英,　澤本和延,　岡野栄之'

実験医学 21 （ 17 ） 2440-2445 2003年11月

研究論文（学術雑誌）, 単著

Symposium 4

Okano Hideyuki, Mizusawa Hidehiro

Clinical Neurology 43 （ 11 ） 823 2003年11月

ISSN 0009-918X

'Origin of higher affinity to RNA of the N-terminal RNA-binding domain than that of the C-terminal one of a mouse neural protein, musashi1, as revealed by comparison of their structures, modes of interaction, surface electrostatic potentials, and backbone dynamics.'

'Miyanoiri Youhei, Kobayashi Hisanori, Imai Takao, Watanabe Michinao, Nagata Takashi, Uesugi Seiichi, Okano Hideyuki<, Katahira Masato'

Journal of Biological Chemistry 278 （ 42 ） 41309-41315 2003年10月

研究論文（学術雑誌）, 共著, 査読有り

F3/Contactin acts as a functional ligand for Notch during oligodendrocyte maturation.

'Hu Qi-Dong, Ang Beng-Ti, Karsak Meliha, Hu Wei-Ping, Cui Xiao-Ying, Duka Tanya, Takeda Yasuo, Chia Wency, Sankar Natesan, Ng Yee-Kong, Ling Eng-Ang, Maciag Thomas, Small Deena, Trifonova Radianna, Kopan Raphael, Okano Hideyuki, Nakafuku Masato, Chiba Shigeru, Hirai Hisamaru, Aster C.Jon, Schachner Melitta, Pallen J.Catherine, Watanabe Kazutada, Xiao Zhi-Cheng'

Cell 115 163-175 2003年10月

研究論文（学術雑誌）, 共著, 査読有り

精神科領域の用語解説・神経幹細胞

'山下徹,　澤本和延,　岡野栄之'

分子精神医学 3 （ 4 ） 342-344 2003年10月

研究論文（学術雑誌）, 単著

Distinct expression patterns of splicing isoforms of mNumb in the endocrine lineage of developing pancreas

'Yoshida Tetsu, Tokunaga Akinori, Nakao Keiko, Okano Hideyuki'

Differentiation 71/8 486-495 2003年10月

研究論文（学術雑誌）, 共著, 査読有り

Cortical and striatal expression of tyrosine hydroxylase mRNA in neonatal and adult mice.

'Harriet Baker, Kobayashi Kazuto, Okano Hideyuki, Saino-Saito Sachiko'

Cellular and Molecular Neurobiology 23/4-5 507-518 2003年10月

研究論文（学術雑誌）, 共著, 査読有り

ES細胞と神経幹細胞の違いとその特性

'吉崎崇仁,　岡野栄之'

Clinical Neuroscience 21 （ 10 ） 1111-1114 2003年10月

研究論文（学術雑誌）, 単著

Hepatocyte growth factor promotes proliferation and neural differentiation of neural stem cells from mouse embryos.

'Kokuzawa Jouji, Yoshimura Shinichi, Kitajima Hideomi, Shinoda Jun, Kaku Yasuhiko, Toru Iwama, Morishita Ryuichi, Shimazaki Takuya, Okano Hideyuki, Kunisada Takahiro, Sakai Noboru'

Molecular and Cellular Neuroscience 24 （ 1 ） 190-197 2003年09月

研究論文（学術雑誌）, 共著, 査読有り

Cytosol-endoplasmic reticulum interplay by Sec61alpha translocon in polyglutamine-mediated neurotoxicity in Drosophila.

'Kanuka Hirotaka, Kuranaga Erina, Tetsuo Hiratou, Igaki Tatsuhi, Nelson Bryce, Okano Hideyuki, Miura Masayuki'

Proceedings　of　the　National　Academy　of　Sciences　of　the　United　States　of　America 100/20 11723-11728 2003年09月

研究論文（学術雑誌）, 共著, 査読有り

Erratum

Hirota Yuki, Sawamoto Kazunobu, Okano Hideyuki

Gene Expression Patterns 3 （ 4 ） 545 - 546 2003年08月

ISSN 1567-133X

Access to Document (DOI)

Caspase-mediated oligodendrocyte cell death in the pathogenesis of autoimmune demyelination.

'Hisahara Shin, Hideyuki Okano, Miura Masayuki'

Neuroscience Research 46 （ 4 ） 387-397 2003年08月

研究論文（学術雑誌）, 共著

神経系におけるRNA結合タンパク質を介した遺伝子発現調節機構

'赤松和土,　岡野栄之'

わかる実験医学シリーズ　RNAがわかる 125-129 2003年07月

研究論文（学術雑誌）, 単著

Academic Press

Okano Hideyuki

Seminars in Cell and Developmental Biology 14 （ 3 ） 159 2003年06月

ISSN 1084-9521

Access to Document (DOI)

Drosophila homeodomain protein REPO controls glial differentiation by cooperating with ETS and BTB transcription factors.

'Yuasa Yoshihiro, Okabe Masataka, Yoshikawa Shingo, Tabuchi Katsuhiko, Xiong Wen-Cheng, Hiromi Yasushi, Okano Hideyuki'

Development 130 2419-2428 2003年06月

研究論文（学術雑誌）, 共著, 査読有り

Proliferation of neural and neuronal progenitors after global brain ischemia in young adult macaque monkeys.

'Tonchev B.Anton, Yamashita Tstumori, Zhao Liang, Okano James Hirotaka, Okano Hideyuki'

Molecular and Cellular Neuroscience 23 （ 2 ） 292-301 2003年06月

研究論文（学術雑誌）, 共著, 査読有り

Making and repairing the mammalian brain: Introduction.

Okano Hideyuki

Seminars in Cell & Developmental Biology 14 （ 3 ） 159 2003年06月

研究論文（学術雑誌）, 単著

Transplantation of neural stem cells into the spinal cord after injury

'Okano Hideyuki, Ogawa Yuto, Nakamura Masaya, Kaneko Shinjiro, Iwanami Akio, Toyama Yoshiaki'

Seminars in Cell & Developmental Biology 14 （ 3 ） 191-198 2003年06月

研究論文（学術雑誌）, 共著

'Differential proliferative response in the postischemic hippocampus, temporal cortex and olfactory bulb of adult primates.'

'Tonchev Anton B, Yamashima Tsutumori, Zhao Liang, Okano Hideyuki'

Glia 42 （ 3 ） 209-224 2003年05月

研究論文（学術雑誌）, 共著, 査読有り

概論・再生医療へ向けた幹細胞研究

岡野栄之

実験医学増刊 21 （ 8 ） 986-989 2003年05月

研究論文（学術雑誌）, 単著

Differentiation of monkey embryonic stem cells into neural lineages.

'Kuo Hung-Chih, Pau Francis K.-Y., Yeoman R. Richard, Mitalipov M. Shoukhrat, Okano Hideyuki, Wolf P.Don'

Biology of Reproduction 68/5 1727-1735 2003年05月

研究論文（学術雑誌）, 共著, 査読有り

神経幹細胞移植による中枢神経系の再生

'岩波明生,　岡野栄之'

細胞工学 22 （ 5 ） 542-547 2003年05月

研究論文（学術雑誌）, 共著

中枢神経系の幹細胞生物学

'神山淳,　岡野栄之'

実験医学増刊 21 （ 8 ） 1020-1026 2003年05月

研究論文（学術雑誌）, 共著

中枢神経系の再生医療

'岡田誠司,　岡野栄之'

再生医療 2 （ 2 ） 89-97 2003年05月

研究論文（学術雑誌）, 共著

Expression of tubulin beta II in neural stem/progenitor cells and radial fibers during human fetal brain development.

'Nakamura Yasuhiro, Yamamoto Munehiko, Oda Eriko, Yamamoto Atsuyo, Kanemura Yonehiro, Hara Masayuki, Okano Hideyuki'

Laboratory Investigation 83 （ 4 ） 479-489 2003年04月

研究論文（学術雑誌）, 共著, 査読有り

神経幹細胞の分化誘導法と中枢神経再生医療

岡田誠司，岡野栄之

日本臨床 61 （ 3 ） 449-456 2003年03月

研究論文（学術雑誌）, 共著

神経系の発生・分化とRNA

岡野栄之，今井貴雄

蛋白質・核酸・酵素　増刊号　RNAの細胞生物学 48 （ 4 ） 430-437 2003年03月

研究論文（学術雑誌）, 共著

Stem Cells Expressing Matrix Metalloproteinase-13 mRNA Appear during Regression Reversal of Hepatic Cirrhosis

Watanabe Tetsu, Niioka Maki, Hozawa Shigenari, Sugioka Yoshihiko, Arai Masao, Maruyama Katsuya, Okano Hideyuki, Okazaki Isao

361 - 388 2003年02月

This chapter describes a mechanism that underlies the degradation of newly formed fibrous bands in CCl<inf>4</inf>-induced liver fibrosis and cirrhosis and demonstrates that neural progenitor cells derived from hematopoietic stem cells express interstitial collagenase in the recovery phase of experimental cirrhosis. It also discusses the availability of hematopoietic stem cells for the treatment of human liver fibrosis or cirrhosis. The reversibility of liver fibrosis in patients with alcoholic cirrhosis and alcoholic fatty liver with fibrosis has been observed after alcohol abstinence. Histologically documented reversibility of liver fibrosis in alcoholic liver diseases was closely correlated with a decrease in serum fibrosis markers of type IV collagen. It is now generally accepted that reversibility of liver fibrosis or cirrhosis occurs under some conditions if the cause of liver damage is removed or adequately treated.

Access to Document (DOI)

神経幹細胞の分化制御機構

松本有史，岡野栄之

ファルマシア 39 （ 2 ） 108-112 2003年02月

研究論文（学術雑誌）, 共著

脊髄損傷に対する再生医療—神経幹細胞移植を中心として

金子慎二郎，中村雅也，岩波明生，小川祐人，岡野栄之，戸山芳昭

脊椎脊髄ジャーナル 16 （ 2 ） 123-129 2003年02月

研究論文（学術雑誌）, 共著

再生医療の現状と将来展望；＜臓器再生＞中枢神経系の再生医療

金子慎二郎，岩波明生，中村雅也，小川祐人，戸山芳昭，岡野栄之

日本医師会雑誌 129 （ 3 ） 355-359 2003年02月

研究論文（学術雑誌）, 共著

中枢神経系の再生医療

金子慎二郎，岩波明生，中村雅也，小川祐人，戸山芳昭，岡野栄之

日本医師会雑誌 129 （ 3 ） 355-359 2003年02月

研究論文（学術雑誌）, 共著

幹細胞を利用した中枢神経系の再生

砂堀毅彦，岡野栄之

最新医学 58 （ 1 ） 102-107 2003年01月

研究論文（学術雑誌）, 共著

Identification of a putative intestinal stem cell and early lineage marker; musashi-1

Potten Christopher S, Booth Catherine, Tudor Gregory L, Booth Dawn, Brady Gerard, Hurley Patricia, Ashton Gary, Clarke Robert, Sakakibara Shin-ichi, Okano Hideyuki

Differentiation 71 （ 1 ） 28-41 2003年01月

研究論文（学術雑誌）, 共著

Candidate markers for stem and early progenitor cells, Musashi-1 and Hes1, are expressed in crypt base columnar cells of mouse small intestine

Kayahara Takahisa, Sawada Mitsutaka, Takaishi Shigeo, Fukui Hirokazu, Seno Hiroshi, Fukuzawa Hiroaki, Suzuki Katsumasa, Hiai Hiroshi, Kageyama Ryoichiro, Okano Hideyuki, Chiba Ttsutomu

FEBS letterers 535 （ 39085 ） 131-135 2003年01月

研究論文（学術雑誌）, 共著

Regulatory mechanisms of neural stem cell and strategies for therapy

Okada Seiji, Okano Hideyuki

Nippon rinsho. Japanese journal of clinical medicine 61 （ 3 ） 449 - 456 2003年

ISSN 0047-1852

Neural stem cells(NSCs) are multipotential progenitor cells that can generate neurons, astrocytes, and oligodendrocytes, the three major cell types in the central nervous system. Due to their self-renewal activities, NSCs can proliferate in an undifferentiated state in vitro, allowing them to be expanded mitotically and harvested in bulk. Recent advances in stem cell biology have led us to investigate methods for the regenerative manipulation of the damaged CNS. However, there is much that is still not known about regulatory mechanisms of the differentiation and self-renewal of NSCs. In this article, we review some of the basic notions regarding the extracellular factors and signal transduction cascades involved in the differentiation and maintenance of NSCs.

Neural stem cells as therapeutic agents for CNS injuries and disorders.

Okano Hideyuki

International Congress Series 1810 39088 2003年

研究論文（学術雑誌）, 単著

Transplantation of neural stem cells into the spinal cord after injury

Okano Hideyuki, Ogawa Yuto, Nakamura Masaya, Kaneko Shinjiro, Iwanami Akio, Toyama Yoshiaki

Seminars in cells & developmental biology 379 39090 2003年

研究論文（学術雑誌）, 共著

Retinal Development and Regeneration

Ozawa Yoko, Oguchi Yoshihisa, Okano Hideyuki

Neuro-Ophthalmology Japan 20 （ 3 ） 272 - 280 2003年

ISSN 0289-7024

One of the final goals of our research is the regaining of normal functions previously lost in damaged tissue. While it has long been believed regeneration of the central nervous system was absolutely impossible, scientists have recently made progress towards this goal. For the regeneration of damaged tissues, both morphological and functional reconstitutions are an indispensable process. As undifferentiated cells, have better survival rates in all types of environments as well as better competency with regard to extracellular signaling factors, we have tried to take advantage of these characteristics. We are trying to find methodologies that will let these cells recapitulate the developmental processes in order to regain the normal physiological functions of damaged tissues. Some of the candidate cells that can be utilized for retinal regeneration are retinal stem cells and embryonic stem cells. In this study, we describe a method for inducing these cells towards specific cell fates as well as their potential therapeutic application. We also describe the significance of investigating the effects of the microenvironment in damaged retina for the purpose of inducing neural retinal regeneration.

RNA regulation in nervous system development and differentiation

Okano Hideyuki, Imai Takao

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 48 （ 4 Suppl ） 430 - 437 2003年

ISSN 0039-9450

Regenerative treatment of nervous system diseases

Matsumoto Arifumi, Okano Hideyuki

Nihon Naika Gakkai zasshi. The Journal of the Japanese Society of Internal Medicine 92 （ 12 ） 2426 - 2432 2003年

ISSN 0021-5384

Differential expression of Pax6 and Ngn2 between pair-generated cortical neurons

Kawaguchi Ayano, Ogawa Masaharu, Saito Kanako, Matsuzaki Fumio, Okano Hideyuki, Miyata Takaki

Journal of Neuroscience Research 78 （ 6 ） 784 - 795 2004年12月

ISSN 0360-4012

Progenitor cells that generate neuron pairs ("pair progenitor cells") are implicated in mammalian cortical development, and their division has been thought to be "symmetric." However, asymmetric growth of two sister neurons generated by the division of a pair progenitor cell would lead to more efficient generation of neuronal diversity in the cortex. To explore mechanisms by which pair progenitor cells provide neuronal diversity, we examined molecular differences between a pair of neurons generated in clonal-density culture. Time-course analysis for the acquisition of neuronal markers and the disappearance of Pax6 and Neurogenin2 (Ngn2) demonstrated that 1) these transcription factors are expressed transiently in some but not all young neurons and 2) some neuron pairs showed uneven/asymmetric expression of Pax6 (19.5%) or Ngn2 (23.8%), whereas other pairs were either symmetrically positive or negative. Asymmetric Pax6 distribution in neuron pairs was not associated with asymmetric distribution of Numb, which raises an intriguing possibility, that Pax6 asymmetry in neuron pairs is produced by an alternative mode of the cell autonomous mechanisms. Stage-dependent changes were noted in the pattern of Ngn2 retention in daughter neurons, reflecting qualitative changes in the pair progenitor population. We suggest that pair progenitor cells contribute to the generation of neuronal diversity through cell-intrinsic heterogeneity and asymmetric division. © 2004 Wiley-Liss, Inc.

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Nonhematopoietic mesenchymal stem cells can be mobilized and differentiate into cardiomyocytes after myocardial infarction

Kawada Hiroshi, Fujita Jun, Kinjo Kentaro, Matsuzaki Yumi, Tsuma Mitsuyo, Miyatake Hiroko, Muguruma Yukari, Tsuboi Kosuke, Itabashi Yuji, Ikeda Yasuo, Ogawa Satoshi, Okano Hideyuki, Hotta Tomomitsu, Ando Kiyoshi, Fukuda Keiichi

Blood 104 （ 12 ） 3581 - 3587 2004年12月

ISSN 0006-4971

Bone marrow (BM) cells are reported to contribute to the process of regeneration following myocardial infarction. However, the responsible BM cells have not been fully identified. Here, we used 2 independent clonal studies to determine the origin of bone marrow (BM)-derived cardiomyocytes. First, we transplanted single CD34^- c-kit⁺Sca-1⁺ lineage^- side population (CD34^-KSL-SP) cells or whole BM cells from mice ubiquitously expressing enhanced green fluorescent protein (EGFP) into lethally irradiated mice, induced myocardial infarction (MI), and treated the animals with granulocyte colony-stimulating factor (G-CSF) to mobilize stem cells to the damaged myocardium. At 8 weeks after MI, from 100 specimens we counted only 3 EGFP⁺ actinin⁺ cells in myocardium of CD34^- KSL-SP cells in mice that received transplants, but more than 5000 EGFP⁺ actinin⁺ cells in whole BM cell in mice that received transplants, suggesting that most of EGFP⁺ actinin⁺ cells were derived from nonhematopoietic BM cells. Next, clonally purified nonhematopoietic mesenchymal stem cells (MSCs), cardiomyogenic (CMG) cells, that expressed EGFP in the cardiomyocyte-specific manner were transplanted directly into BM of lethally irradiated mice, MI was induced, and they were treated with G-CSF. EGFP⁺ actinin⁺ cells were observed in the ischemic myocardium, indicating that CMG cells had been mobilized and differentiated into cardiomyocytes. Together, these results suggest that the origin of the vast majority of BM-derived cardiomyocytes is MSCs. © 2004 by The American Society of Hematology.

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Transcriptional integration of cell polarity and positional identity specifies daughters cell-fates in asymmetric cell division

岡野栄之

ZOOLOGICAL SCIENCE 21 （ 12 ） 1280 - 1280 2004年12月

研究論文（学術雑誌）, ISSN 0289-0003

Retinoic-acid-concentration-dependent acquisition of neural cell identity during in vitro differentiation of mouse embryonic stem cells

Okada Yohei, Shimazaki Takuya, Sobue Gen, Okano Hideyuki

Developmental Biology 275 （ 1 ） 124 - 142 2004年11月

ISSN 0012-1606

Retinoic acid (RA) is one of the most important morphogens, and its embryonic distribution correlates with neural differentiation and positional specification in the developing central nervous system. To investigate the concentration-dependent effects of RA on neural differentiation of mouse embryonic stem cells (ES cells), we investigated the precise expression profiles of neural and regional specific genes by ES cells aggregated into embryoid bodies (EBs) exposed to various concentrations of RA or the BMP antagonist Noggin. RA promoted both neural differentiation and caudalization in a concentration-dependent manner, and the concentration of RA was found to regulate dorso-ventral identity, i.e., higher concentrations of RA induced a dorsal phenotype, and lower concentrations of RA induced a more ventral phenotype. The induction of the more ventral phenotype was due to the higher expression level of the N-terminus of sonic hedgehog protein (Shh-N) when treated with low concentration RA, as it was abrogated by an inhibitor of Shh signaling, cyclopamine. These findings suggest that the concentration of RA strictly and simultaneously regulates the neuralization and positional specification during differentiation of mouse ES cells and that it may be possible to use it to establish a strategy for controlling the identity of ES-cell-derived neural cells. © 2004 Elsevier Inc. All rights reserved.

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Drosophila deltex mediates suppressor of hairless-independent and late-endosomal activation of notch signaling

Hori Kazuya, Fostier Maggy, Ito Mikiko, Fuwa Takashi J., Go Masahiro J., Okano Hideyuki, Baron Martin, Matsuno Kenji

Development 131 （ 22 ） 5527 - 5537 2004年11月

ISSN 0950-1991

Notch (N) signaling is an evolutionarily conserved mechanism that regulates many cell-fate decisions. deltex (dx) encodes an E3-ubiquitin ligase that binds to the intracellular domain of N and positively regulates N signaling. However, the precise mechanism of Dx action is unknown. Here, we found that Dx was required and sufficient to activate the expression of gene targets of the canonical Su(H)-dependent N signaling pathway. Although Dx required N and a cis-acting element that overlaps with the Su(H)-binding site, Dx activated a target enhancer of N signaling, the dorsoventral compartment boundary enhancer of vestigial (vgBE), in a manner that was independent of the Delta (DI)/Serrate (Ser) ligands- or Su(H). Dx caused N to be moved from the apical cell surface into the late-endosome, where it accumulated stably and co-localized with Dx. Consistent with this, the dx gene was required for the presence of N in the endocytic vesicles. Finally, blocking the N transportation from the plasma membrane to the late-endosome by a dominant-negative form of Rab5 inhibited the Dx-mediated activation of N signaling, suggesting that the accumulation of N in the late-endosome was required for the Dx-mediated Su(H)-independent N signaling.

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Human neural stem/progenitor cells, expanded in long-term neurosphere culture, promote functional recovery after focal ischemia in Mongolian gerbils

Ishibashi Satoru, Sakaguchi Masanori, Kuroiwa Toshihiko, Yamasaki Mami, Kanemura Yonehiro, Shizuko Ichinose, Shimazaki Takuya, Onodera Masafumi, Okano Hideyuki, Mizusawa Hidehiro

Journal of Neuroscience Research 78 （ 2 ） 215 - 223 2004年10月

ISSN 0360-4012

Transplantation of human neural stem cells (NSCs) is a promising potential therapy for neurologic dysfunctions after the hyperacute stage of stroke in humans, but large amounts of human NSCs must be expanded in long-term culture for such therapy. To determine their possible therapeutic potential for human stroke, human fetal neural stem/progenitor cells (NSPCs) (i.e., neurosphere-forming cells) were isolated originally from forebrain tissues of one human fetus, and expanded in long-term neurosphere culture (exceeding 24 weeks), then xenografted into the lesioned areas in the brains of Mongolian gerbils 4 days after focal ischemia. Sensorimotor and cognitive functions were evaluated during the 4 weeks after transplantation. The total infarction volume in the NSPC-grafted animals was significantly lower than that in controls. Approximately 8% of the grafted NSPCs survived, mainly in areas of selective neuronal death, and were costained with antibodies against neuronal nuclei antibody (NeuN), microtubule associated protein (MAP-2), glial fibrillary acidic protein (GFAP), and anti-2′3′ cyclic nucleotide 3′-phosphodiesterase (CNPase). Synaptic structures between NSPCs-derived neurons and host neurons were observed. Furthermore, gradual improvement of neurologic functions was observed clearly in the NSPC-grafted animals, compared to that in controls. Human NSPCs, even from long-term culture, remarkably improved neurologic functions after focal ischemia in the Mongolian gerbil, and maintained their abilities to migrate around the infarction, differentiate into mature neurons, and form synapses with host neuronal circuits. These results indicate that in vitro-expanded human neurosphere cells are a potential source for transplantable material for treatment of stroke. © 2004 Wiley-Liss, Inc.

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Human amnion mesenchyme cells express phenotypes of neuroglial progenitor cells

Sakuragawa Norio, Kakinuma Kenichi, Kikuchi Aiko, Okano Hideyuki, Uchida Saiko, Kamo Isao, Kobayashi Mamoru, Yokoyama Yasunobu

Journal of Neuroscience Research 78 （ 2 ） 208 - 214 2004年10月

ISSN 0360-4012

Previous studies from our laboratory showed that human amnion epithelial cells (AECs) have multiple functions, such as synthesis and release of catecholamines, acetylcholine, neurotrophic factors, activin, and noggin. In this study, we investigated the identity of neural progenitor cells in human amnion mesenchyme cells (AMCs), which lie immediately adjacent to the AECs. Cryostat sections revealed that vimentin expression was detected in the AMCs and CK19 in AECs. Vimentin-positive cells made up 97.5% of total cells tested in cultured AMCs. Interestingly, 3.6% of total AMCs expressed the phenotype CK19+/vimentin +, indicating coexpression of epithelial and mesenchyme cell markers. In culturing with bromodeoxyuridine (BrdU) for 24 hr, 66-82% of cells were found to be BrdU positive, suggesting that they have proliferating potency. By using RT-PCR, AMCs express mRNA of nestin and Musashi1. With a neural cell differentiating protocol, cell bodies extended long bipolar or complex multipolar processes. Nestin (87.7% of total cells tested) and Musashi1 (93.1%) were expressed in undifferentiated cells, and their positively stained cells increased in number slightly after induction. Undifferentiated cells were stained by anti-Tuj1 and NF-M, and their positively stained cells increased significantly in number after induction, to 72.8% and 46.0%, respectively. Meanwhile, glial fibrillary acidic protein-positive cells increased from 25.4% to 43.2% after induction. These studies demonstrate that AMCs have phenotypes of neuroglial progenitor cells and can be differentiated into neuroglial phenotypes by optimal differentiation protocol. Eventually, AMC-derived stem cells may be a favorable cell vehicle in regenerative medicine. © 2004 Wiley-Liss, Inc.

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Possible oncogenicity of subventricular zone neural stem cells

Uchida Koichi, Mukai Makio, Okano Hideyuki, Kawase Takeshi

Neurosurgery 55 （ 4 ） 977 - 978 2004年10月

ISSN 0148-396X

OBJECTIVE: The origin of brain tumors has attracted much controversy. Recent advances in neural stem cell biology coupled with the new concepts of central nervous system development have raised interesting possibilities regarding the oncogenic properties of neural stem/progenitor cells. To elucidate these putative properties further, the clinical materials from an infant brain tumor were analyzed, focusing on the relation with the neural stem/progenitor cells. METHODS: The expression of neural stem/progenitor cell markers in the tumor cells and the cellular components of the infant brain tumor were examined using immunohistochemistry. The tumor cell biology was analyzed both in culture and in the grafted brain environment. RESULTS: Three main bodies of evidence were demonstrated indicating that the tumor was of possible subventricular zone postnatal or adult normal neural stem cell origin. First, in the tumor specimen we demonstrated the strong positive expression of the neural stem/progenitor cell markers, nestin and Musashi-1. Second, immunohistochemistry revealed the presence of neuronal, astrocytic, and immature precursor cells in the tumor tissue, similar to the cellular components of the subventricular zone, thereby pointing to the subventricular zone as the possible origin of the tumor. The subventricular zone also is one of the strong candidates for the location of postnatal/adult neural stem cells. This cellular evidence was strengthened further by the clinicoradiological findings that demonstrated the involvement of the subventricular zone of the lateral ventricle by the tumor. Third, in the in vitro and in vivo experiments, a dynamic shift in expression patterns between neural stem cells (nestin, Musashi-1) and differentiated cells (glial fibrillary acidic protein, neuron-specific Class III β-tubulin) markers was seen, similar to the proposed behavior of postnatal/adult neural stem cells in situ. CONCLUSION: These findings suggest that this brain tumor originated from neural stem cells located in the subventricular zone, and the further possibility of the general oncogenic potential of neural stem cells.

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Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors

Unezaki Sawako, Nishizawa Mikio, Okuda-Ashitaka Emiko, Masu Yasuo, Mukai Masanori, Kobayashi Satoru, Sawamoto Kazunobu, Okano Hideyuki, Ito Seiji

Gene 336 （ 1 ） 47 - 58 2004年07月

ISSN 0378-1119

We previously described two isoforms (MOVO-A and -B) of a novel zinc finger protein MOVO, a mouse homologue of Drosophila Ovo protein. Here, we isolated cDNA encoding the third isoform MOVO-C, which had a transactivation domain and zinc finger domain, but lacked an N-terminal potential repression domain that was present in MOVO-A. Three isoform mRNAs were expressed highly in mouse testis and also in the ovary at lower levels. The structural analyses of the isolated Movo gene and mRNAs demonstrated that three different Movo transcripts were differentially processed to generate three isoforms. Major mRNA species encoded MOVO-B with a zinc finger domain alone, and minor mRNA species encoded MOVO-A (potential repressor) and MOVO-C (potential activator). To assign MOVO to a transcriptional factor, we characterized DNA-binding and transactivation properties. Random oligonucleotide selection, electrophoretic mobility shift assay and footprinting indicated that MOVO bound to the sequence, 5′-G(G/C/T)GGGGG-3′. These motifs were found in the 5′-flanking regions of Movo and other testis-specific genes. Nuclear proteins binding to this motif were detected in mouse testis, and the expression of MOVO mRNA was restricted in spermatocytes. The luciferase assay demonstrated that MOVO-C activated Movo promoter and MOVO-A repressed it, but MOVO-B had no effects. Mutated MOVO-binding motifs in the Movo promoter reduced the luciferase activity. All the isoforms had no effects on SV40 promoter without MOVO-binding motifs. MOVO-A partially rescued oogenesis of a Drosophila ovo mutant. These results suggest that MOVO isoforms are transcription factors to regulate genes carrying the MOVO-binding motifs in the testis. © 2004 Elsevier B.V. All rights reserved.

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Mapping spatio-temporal activation of Notch signaling during neurogenesis and gliogenesis in the developing mouse brain

Tokunaga Akinori, Kohyama Jun, Yoshida Tetsu, Nakao Keiko, Sawamoto Kazunobu, Okano Hideyuki

Journal of Neurochemistry 90 （ 1 ） 142 - 154 2004年07月

ISSN 0022-3042

Notch1 plays various important roles including the maintenance of the stem cell state as well as the promotion of glial fates in mammalian CNS development. However, because of the very low amount of the activated form of Notch1 present in vivo, its precise activation pattern has remained unknown. In this study, we mapped the active state of this signaling pathway in situ in the developing mouse brain using a specific antibody that recognizes the processed form of the intracellular domain of Notch1 cleaved by presenilin/γ-secretase activity. By using this antibody, active state of Notch1 came to be detectable with a higher sensitivity than using conventional antibody against Notch1. We found that activated Notch1 was mainly detected in the nuclei of a subpopulation of radial glial cells, the majority of proliferating precursor cells in the ventricular zone (VZ). However, Notch1 activation was not detected in neuronal precursor cells positive for neuronal basic helix-loop-helix proteins or in differentiating neurons in the embryonic forebrain. Interestingly, we found that Notch1 was transiently activated in the astrocytic lineage during perinatal CNS development. Taken together, the present method has enabled us to determine the timing, gradients, and boundaries of the activation of Notch signaling.

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Isolation and transplantation of dopaminergic neurons generated from mouse embryonic stem cells

Yoshizaki Takahito, Inaji Motoki, Kouike Hiroko, Shimazaki Takuya, Sawamoto Kazunobu, Ando Kiyoshi, Date Isao, Kobayashi Kazuto, Suhara Tetsuya, Uchiyama Yasuo, Okano Hideyuki

Neuroscience Letters 363 （ 1 ） 33 - 37 2004年06月

ISSN 0304-3940

Embryonic stem (ES) cells differentiate into dopamine (DA)-producing neurons when co-cultured with PA6 stromal cells, but the resulting cultures contain a variety of unidentified cells. In order to label live DA neurons in mixed populations, we introduced a GFP reporter under the control of the tyrosine hydroxylase (TH) gene promoter into ES cells. GFP expression was observed in TH-immunoreactive cells that differentiated from the ES cells that carried the TH-GFPreporter gene. DA neurons expressing GFP were sorted from the mixed cell population by fluorescence-activated cell sorting of cells exhibiting GFP fluorescence, and the sorted GFP⁺ cells obtained were transplanted into a rat model of Parkinson's disease. Some of these cells survived and innervated the host striatum, resulting in a partial recovery from parkinsonian behavioral defects. This strategy of isolation and transplantation of ES-cell-derived DA neurons should be useful for cellular and molecular studies of DA neurons and for clinical application in the treatment of Parkinson's disease. © 2004 Elsevier Ireland Ltd. All rights reserved.

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Downregulation of STAT3 activation is required for presumptive rod photoreceptor cells to differentiate in the postnatal retina

Ozawa Yoko, Nakao Keiko, Shimazaki Takuya, Takeda Junji, Akira Shizuo, Ishihara Katsuhiko, Hirano Toshio, Oguchi Yoshihisa, Okano Hideyuki

Molecular and Cellular Neuroscience 26 （ 2 ） 258 - 270 2004年06月

ISSN 1044-7431

Ciliary neurotrophic factor (CNTF) has been known to inhibit the differentiation of presumptive rod photoreceptor cells; however, the underlying mechanisms have remained to be elucidated. We demonstrated that STAT3 activation, but not SHP2 activation, is responsible for the CNTF/gp130 signaling that inhibits expression of Rhodopsin and its upstream activator, crx, in the retinal explants derived from P0 mice (P0 retinal explants), utilizing STAT3-deficient retina and electroporation of dominant-negative form of STAT3 (STAT3F). We also demonstrated that STAT3 activation in presumptive rod photoreceptor cells at E18.5 is rapidly downregulated at P0, when Rhodopsin expression starts during retinal development. Persistent STAT3 activation in the P0 retinal explants prevented Rhodopsin expression and rapid upregulation of crx expression. STAT3-deficient retinas did not exhibit precocious rod photoreceptor cell differentiation as a whole, although they occasionally exhibited precocious upregulation of crx mRNA. Thus, we conclude that downregulation of STAT3 activation is required, but insufficient, for rod photoreceptor cell differentiation in the postnatal retina. © 2004 Elsevier Inc. All rights reserved.

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Implantation of Dendritic Cells in Injured Adult Spinal Cord Results in Activation of Endogenous Neural Stem/Progenitor Cells Leading to De Novo Neurogenesis and Functional Recovery

Mikami Yuji, Okano Hideyuki, Sakaguchi Masanori, Nakamura Masaya, Shimazaki Takuya, Okano Hirotaka James, Kawakami Yutaka, Toyama Yoshiaki, Toda Masahiro

Journal of Neuroscience Research 76 （ 4 ） 453 - 465 2004年05月

ISSN 0360-4012

We report a treatment for spinal cord injury involving implantation of dendritic cells (DCs), which act as antigen-presenting cells in the immune system. The novel mechanisms underlying this treatment produce functional recovery. Among the immune cells tested, DCs showed the strongest activity inducing proliferation and survival of neural stem/progenitor cells (NSPCs) in vitro. Furthermore, in DC-implanted adult mice, endogenous NSPCs in the injured spinal cord were activated for mitotic de novo neurogenesis. These DCs produced neurotrophin-3 and activated endogenous microglia in the injured spinal cord. Behavioral analysis revealed the locomotor functions of DC-implanted mice to have recovered significantly as compared to those of control mice. Our results suggest that DC-implantation exerts trophic effects, including activation of endogenous NSPCs, leading to repair of the injured adult spinal cord. © 2004 Wiley-Liss, Inc.

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RMD-1 is a novel protein with multiple coiled-coil domains that regulates microtubule-based processes in C. elegans embryos

岡野栄之

CELL STRUCTURE AND FUNCTION 29 36 - 36 2004年05月

研究論文（学術雑誌）, ISSN 0386-7196

Spatiotemporal patterns of Musashil expression during inner ear development

Sakaguchi Hirofumi, Yaoi Takeshi, Suzuki Toshihiro, Okano Hideyuki, Hisa Yasuo, Fushiki Shinji

NeuroReport 15 （ 6 ） 997 - 1001 2004年04月

ISSN 0959-4965

Musashil (Msi I) is an RNA binding protein associated with asymmetric cell divisions in neural progenitor cells.To investigate the involvement of MsiI in the inner ear development, we studied the expression of MsiI in mouse inner ears with RT-PCR and immunohistochemistry. Immunohistochemistry revealed that MsiI was expressed in all otocyst cells at embryonic day (E) 10 and 12. MsiI immunoreactivity became lost in hair cells after E14 in vestibule and after E16 in cochlea, whereas it persisted in supporting cells until adulthood. The subcellular localization of MsiI changed from "cytoplasmic predominance" to "nuclear predominance" during the first 2 weeks after birth. The present data suggested that Msi may play a role in inner ear development. © 2004 Lippincott Williams & Wilkins.

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Blockade of Interleukin-6 Receptor Suppresses Reactive Astrogliosis and Ameliorates Functional Recovery in Experimental Spinal Cord Injury

Okada S., Nakamura M., Mikami Y., Shimazaki T., Mihara M., Ohsugi Y., Iwamoto Y., Yoshizaki K., Kishimoto T., Toyama Y., Okano Hideyuki

Journal of Neuroscience Research 76 （ 2 ） 265 - 276 2004年04月

ISSN 0360-4012

Endogenous neural stem/progenitor cells (NSPCs) have recently been shown to differentiate exclusively into astrocytes, the cells that are involved in glial scar formation after spinal cord injury (SCI). The microenvironment of the spinal cord, especially the inflammatory cytokines that dramatically increase in the acute phase at the injury site, is considered to be an important cause of inhibitory mechanism of neuronal differentiation following SCI. Interleukin-6 (IL-6), which has been demonstrated to induce NSPCs to undergo astrocytic differentiation selectively through the JAK/STAT pathway in vitro, has also been demonstrated to play a critical role as a proinflammatory cytokine and to be associated with secondary tissue damage in SCI. In this study, we assessed the efficacy of rat anti-mouse IL-6 receptor monoclonal antibody (MR16-1) in the treatment of acute SCI in mice. Immediately after contusive SCI with a modified NYU impactor, mice were intraperitoneally injected with a single dose of MR16-1 (100 μg/g body weight), the lesions were assessed histologically, and the functional recovery was evaluated. MR16-1 not only suppressed the astrocytic diffentiation-promoting effect of IL-6 signaling in vitro but inhibited the development of astrogliosis after SCI in vivo. MR16-1 also decreased the number of invading inflammatory cells and the severity of connective tissue scar formation. In addition, we observed significant functional recovery in the mice treated with MR16-1 compared with control mice. These findings suggest that neutralization of IL-6 signaling in the acute phase of SCI represents an attractive option for the treatment of SCI. © 2004 Wiley-Liss, Inc.

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Sox21 is a repressor of neuronal differentiation and is antagonized by YB-1

Ohba Hiroyuki, Chiyoda Tatsuyuki, Endo Emmy, Yano Masato, Hayakawa Yoshika, Sakaguchi Masanori, Darnell Robert B., Okano Hirotaka J., Okano Hideyuki

Neuroscience Letters 358 （ 3 ） 157 - 160 2004年04月

ISSN 0304-3940

Sox21, a high-mobility group box transcription factor, is expressed throughout the immature neural stem/progenitor population in ventricular zone but not in the cortical plate in embryonic mouse brain and its expression is restricted to the subventricular zone in the adult brain. In undifferentiated PC12 cells, endogenous Sox21 expression is detected but its expression ceases during the nerve growth factor (NGF)-induced neuronal differentiation. Overexpression of Sox21 results in a substantial repression of NGF-induced neurite outgrowth in PC12 cells. Further, we biochemically identified a Sox21-associating protein, Y-box binding protein 1 which not only binds to Sox21 but also partially restores NGF-induced neurite outgrowth of PC12 cells inhibited by Sox21. These results suggest that Sox21 is a repressor of neuronal differentiation in the developing nervous system. © 2004 Elsevier Ireland Ltd. All rights reserved.

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再生医療の過去、現在、未来

岡野栄之

炎症・再生 24 （ 2 ） 82-83 2004年03月

研究論文（学術雑誌）, 単著

Endothelin-1 regulates cardiac sympathetic innervation in the rodent heart by controlling nerve growth factor expression.

'Ieda Masaki, Fukuda Keiichi, Hisaka Yasuyo, Kimura Kensuke, Kawaguchi Haruko, Fujita Jun, Shimida Kouji, Takeshita Eiko, Okano Hideyuki, Kurihara Yukiko, Kurihara Hiroki, Ishida Junji, Fukamizu Akiyoshi, Federoff J. Howard, Ogawa Satoshi'

Journal of Clinical Investigation 113 876-884 2004年03月

研究論文（学術雑誌）, 共著, 査読有り

幹細胞を用いた移植治療

'辻俊幸,　吉崎崇仁,　岡野栄之'

分子リウマチ 1 （ 1 ） 50-55 2004年02月

研究論文（学術雑誌）, 単著

Expression of Musashi1, a neural RNA-binding protein, in the cochlea of young adult mice

Murata Junko, Murayama Ayako, Horii Arata, Doi Katsumi, Harada Tamotsu, Okano Hideyuki, Kubo Takeshi

Neuroscience Letters 354 （ 3 ） 201 - 204 2004年01月

ISSN 0304-3940

Musashi1 (Msi1) is an RNA-binding protein expressed in neural stem/progenitor cells, astroglial progenitor cells and astrocytes in the vertebrate central nervous system. We hypothesized that Msi1 is expressed in only some of the supporting cells in the cochlea, which could become hair cell progenitors under special circumstances after an injury. To observe this, we investigated Msi1 expression in young adult mouse cochlea by immunohistochemistry using monoclonal antibody against Msi1. Msi1 immunostaining was found in a variety of supporting cells but not in outer hair cells in the organ of Corti. Although an immunoreactive ring was found around the inner hair cells, it also seemed to originate from the supporting cells. We suppose that this wide expression of Msi1 in supporting cells indicates that those cells might have the potential to become hair cell progenitors if injured, but that some other mechanisms strictly inhibit this ability. © 2003 Elsevier Ireland Ltd. All rights reserved.

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Unexpectedly efficient homing capacity of purified murine hematopoietic stem cell.

'Matsuzaki Yumi, Kinjo Kentaro, Mulligan C.Richard, Okano Hideyuki'

Immunity 20 87-93 2004年01月

研究論文（学術雑誌）, 共著, 査読有り

Comparison between fetal spinal-cord- and forebrain-derived neural stem/progenitor cells as a source of transplantation for spinal cord injury

Watanabe Kota, Nakamura Masaya, Iwanami Akio, Fujita Yuko, Kanemura Yonehiro, Toyama Yoshiaki, Okano Hideyuki

Developmental Neuroscience 26 （ 2-4 ） 275 - 287 2004年

ISSN 0378-5866

Recently, we have shown that the transplantation of spinal-cord-derived neural stem/progenitor cells (NSPCs) can contribute to the repair of injured spinal cords in adult rats, which may correspond to a behavioral recovery. To apply these results to clinical practice, a system for supplying human NSPCs on a large scale must be established. However, human spinal-cord-derived NSPCs are known to have a low proliferation rate, compared with forebrain-derived NSPCs. This low proliferative potency limits the feasibility of large-scale spinal cord-derived NSPC use. Thus, forebrain-derived NSPCs should be examined as an alternative to spinal-cord-derived NSPCs for the treatment of spinal cord injuries. In this study, we compared spinal-cord- and forebrain-derived NSPCs transplanted into injured spinal cords with respect to their fates in vivo as well as the animals' functional recovery. Both spinal-cord- and forebrain-derived NSPCs promoted functional recovery in rats with spinal cord injuries. While both spinal-cord- and fore-brain-derived NSPCs survived, migrated and differentiated into neurons, astrocytes and oligodendrocytes in response to the microenvironment within the injured spinal cord after transplantation, forebrain-derived NSPCs differentiated into more neurons and fewer oligodendrocytes, compared to spinal-cord-derived NSPCs. Neurons that had differentiated from the transplanted forebrain-derived NSPCs were shown to be positive for neurotransmitters like GABA, glutamate and glycine, although authentic glycinergic neurons are not normally present within the forebrain. Thus, at least a subpopulation of the transplanted forebrain-derived NSPCs differentiated into spinal-cord-type neurons. In conclusion, forebrain-derived NSPCs could be used as an alternative to spinal-cord-derived NSPCs as a potential therapeutic agent for spinal cord injuries. Copyright © 2004 S. Karger AG.

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Vascular adventitia generates neuronal progenitors in the monkey hippocampus after ischemia

Yamashima Tetsumori, Tonchev Anton B., Vachkov Ivan H., Popivanova Boryana K., Seki Tatsunori, Sawamoto Kazunobu, Okano Hideyuki

Hippocampus 14 （ 7 ） 861 - 875 2004年

ISSN 1050-9631

In the adult hippocampus, neurogenesis proceeds in the subgranular zone (SGZ) of the dentate gyrus (DG), but not in the cornu Ammonis (CA). Recently, we demonstrated in monkeys that transient brain ischemia induces an increase of the neuronal progenitor cells in the SGZ, but not in CA1, in the second week after the insult. To identify the origin of primary neuronal progenitors in vivo, we compared the postischemic monkey DG and CA1, using light and electron microscopy, focusing on specific phenotype markers, as well as the expression of neurotrophic factors. Laser confocal microscopy showed that 1-3% of 5-bromo-2′-deoxyuridine (BrdU)-positive cells in the SGZ after 2-96 h labeling were also positive for neuronal markers such as TUC4, βIII tubulin, and NeuN on days 9 and 15. In contrast, despite the presence of numerous BrdU-positive cells, CA1 showed no neurogenesis at any time points, and all the progenitors were positive for glial markers: Iba1 or S-100β on days 4, 9, and 15. Highly polysialylated neural cell adhesion molecule (PSA-NCAM)-positive cells were abundant in the SGZ, but were absent in CA1. On day 9, most of the immature neurons positive for βIII-tubulin in SGZ showed an increase in PSA-NCAM immunoreactivity. The immunoreactivity of brain-derived neurotrophic factor (BDNF) was abundant at the vascular adventitia of the SGZ, but was absent at the adventitia of CA1. BrdU-positive progenitor cells were frequently seen in the vicinity of proliferating blood vessels. Ultrastructural analysis indicated that most of the neuronal progenitor cells and microglia originated from the pericytes of capillaries and/or adventitial cells of arterioles (called vascular adventitia). The detaching adventitial cells showed mitotic figures in the perivascular space, and the resultant neuronal progenitor cells made contact with dendritic spines associated with synaptic vesicles or boutons. These data implicate the vascular adventitia as a novel potential source of neuronal progenitor cells in the postischemic primate SGZ. © 2004 Wiley-Liss, Inc.

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Correlation between quantitative imaging and behavior in unilaterally 6-OHDA-lesioned rats

Inaji Motoki, Okauchi Takashi, Ando Kiyoshi, Maeda Jun, Nagai Yuji, Yoshizaki Takahito, Okano Hideyuki, Nariai Tadashi, Ohno Kikuo, Obayashi Shigeru, Higuchi Makoto, Suhara Tetsuya

Brain Research 1064 （ 1-2 ） 136 - 145 2005年12月

ISSN 0006-8993

We evaluated correlation between neurochemical and functional alterations of the nigrostriatal dopaminergic system in rat brains lesioned with 6-hydroxydopamine (6-OHDA), that model hemi-Parkinson's disease (PD), by using three different quantitative in vivo and in vitro methods. Rats unilaterally lesioned with different doses of 6-OHDA underwent two types of in vivo experiments: (1) a rotational behavioral study with methamphetamine (MAP) or apomorphine (APO); and (2) a positron emission tomography (PET) study with [¹¹C]PE2I (radioligand for dopamine transporters) or [ ¹¹C]raclopride (radioligand for dopamine D2 receptors). An in vitro autoradiographic study with the same radioligands was also conducted. The number of rotations after the MAP or APO injection increased with increased doses of 6-OHDA. The in vitro and in vivo binding of [¹¹C]PE2I dose-dependently decreased in response to the 6-OHDA injections, while that of [¹¹C]raclopride dose-dependently increased. There was a significant negative hyperbolic correlation between the number of rotations after MAP injection and the binding of [¹¹C]PE2I. In contrast, there was a significant positive linear correlation between the number of rotations after APO injections and the binding of [¹¹C]raclopride. These results robustly reveal a molecular pharmacological basis of parkinsonian symptoms in animal models of PD, and indicate the utility and validity of in vivo PET measurements in assessing pre- and post-synaptic dopaminergic functions. © 2005 Elsevier B.V. All rights reserved.

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Chondroitinase ABC combined with neural stem/progenitor cell transplantation enhances graft cell migration and outgrowth of growth-associated protein-43-positive fibers after rat spinal cord injury

Ikegami Takeshi, Nakamura Masaya, Yamane Junichi, Katoh Hiroyuki, Okada Seiji, Iwanami Akio, Watanabe Kota, Ishii Ken, Kato Fumikazu, Fujita Hiroshi, Takahashi Toyomi, Okano Hirotaka James, Toyama Yoshiaki, Okano Hideyuki

European Journal of Neuroscience 22 （ 12 ） 3036 - 3046 2005年12月

ISSN 0953-816X

We previously reported that the transplantation of neural stem/progenitor cells (NSPCs) can contribute to the repair of injured spinal cord in adult rats and monkeys. In some cases, however, most of the transplanted cells adhered to the cavity wall and failed to migrate and integrate into the host spinal cord. In this study we focused on chondroitin sulfate proteoglycan (CSPG), a known constituent of glial scars that is strongly expressed after spinal cord injury (SCI), as a putative inhibitor of NSPC migration in vivo. We hypothesized that the digestion of CSPG by chondroitinase ABC (C-ABC) might promote the migration of transplanted cells and neurite outgrowth after SCI. An in vitro study revealed that the migration of NSPC-derived cells was inhibited by CSPG and that this inhibitory effect was attenuated by C-ABC pre-treatment. Consistently, an in vivo study of C-ABC treatment combined with NSPC transplantation into injured spinal cord revealed that C-ABC pre-treatment promoted the migration of the transplanted cells, whereas CSPG-immunopositive scar tissue around the lesion cavity prevented their migration into the host spinal cord in the absence of C-ABC pre-treatment. Furthermore, this combined treatment significantly induced the outgrowth of a greater number of growth-associated protein-43-positive fibers at the lesion epicentre, compared with NSPC transplantation alone. These findings suggested that the application of C-ABC enhanced the benefits of NSPC transplantation for SCI by reducing the inhibitory effects of the glial scar, indicating that this combined treatment may be a promising strategy for the regeneration of injured spinal cord. © Federation of European Neuroscience Societies.

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Downregulation of Notch, mediated by numb and -adaptin, is required for the differentiation of postembryonic neural stem cells in Drosophila

岡野栄之

ZOOLOGICAL SCIENCE 22 （ 12 ） 1468 - 1468 2005年12月

研究論文（学術雑誌）, ISSN 0289-0003

Gain-of-function screen identifies a role of the Sec61α translocon in Drosophila postmitotic neurotoxicity

Kanuka Hirotaka, Hiratou Tetsuo, Igaki Tatsushi, Kanda Hiroshi, Kuranaga Erina, Sawamoto Kazunobu, Aigaki Toshiro, Okano Hideyuki, Miura Masayuki

Biochimica et Biophysica Acta - General Subjects 1726 （ 3 ） 225 - 237 2005年11月

ISSN 0304-4165

To elucidate the intrinsic mechanisms of neurotoxicity induction, including those underlying neural cell death and neurodegeneration, we developed a gain-of-function screen for gene products causing neural cell loss. To identify novel genes with a cell-death-related function in neurons, we screened 4,964 Drosophila GS lines, in which one or two genes from much of the Drosophila genome can be overexpressed. Approximately 0.68% of the GS lines produced phenotypes involving a loss of postmitotic neurons. Of these, we identified and characterized the endd2 gene, which encodes the Drosophila ortholog of Sec61α (DSec61α), an endoplasmic reticulum protein with protein translocation activity. Ectopic expression of DSec61α caused neural cell death accompanied by the accumulation of ubiquitinated proteins, which was mediated by DSec61α's translocon activity. This supported our previous observation that the DSec61α translocon contributes to expanded polyglutamine-mediated neuronal toxicity, which is also associated with ubiquitinated protein accumulation. These data suggest that the translocon may be a novel component of neural cell death and degeneration pathways. Our approach can be used to identify potential neurotoxic factors within the whole genome, which will increase our understanding of the molecular mechanisms of various types of cell death, including those associated with human neurodegenerative diseases. © 2005 Elsevier B.V. All rights reserved.

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Hzf protein regulates dendritic localization and BDNF-induced translation of type 1 inositol 1,4,5-trisphosphate receptor mRNA

Iijima Takatoshi, Imai Takao, Kimura Yuki, Bernstein Alan, Okano Hirotaka James, Yuzaki Michisuke, Okano Hideyuki

Proceedings of the National Academy of Sciences of the United States of America 102 （ 47 ） 17190 - 17195 2005年11月

ISSN 0027-8424

The localization of certain mRNAs to dendrites and their local translation in synaptic regions are proposed to be involved in certain aspects of synaptic plasticity. A cis-acting element within the 3′ untranslated region (3′ UTR) of the targeted mRNAs, which is bound by a trans-acting RNA-binding protein, controls the dendritic mRNA localization. Here, we identified hematopoietic zinc finger (Hzf) as a trans-acting factor that regulates the dendritic mRNA localization of the type 1 inositol 1,4,5-trisphosphate receptor (IP₃RI), a dendritically localized mRNA in cerebellar Purkinje cells, via binding to the 3′ UTR. In Hzf-deficient mice, the dendritic localization of IP₃RI mRNA and brain-derived neurotrophic factor-induced IP₃RI protein synthesis in the cerebellum were impaired. These findings suggest that Hzf is an RNA-binding protein that controls the dendritic mRNA localization and activity-dependent translation of IP₃RI, and may be involved in some aspects of synaptic plasticity. © 2005 by The National Academy of Sciences of the USA.

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Drosophila caspase transduces Shaggy/GSK-3β kinase activity in neural precursor development

Kanuka Hirotaka, Kuranaga Erina, Takemoto Kiwamu, Hiratou Tetsuo, Okano Hideyuki, Miura Masayuki

EMBO Journal 24 （ 21 ） 3793 - 3806 2005年11月

ISSN 0261-4189

All Rights Reserved.Caspases are well known for their role in the execution of apoptotic programs, in which they cleave specific target proteins, leading to the elimination of cells, and for their role in cytokine maturation. In this study, we identified a novel substrate, which, through cleavage by caspases, can regulate Drosophila neural precursor development. Shaggy (Sgg)46 protein, an isoform encoded by the sgg gene and essential for the negative regulation of Wingless signaling, is cleaved by the Dark-dependent caspase. This cleavage converts it to an active kinase, which contributes to the formation of neural precursor (sensory organ precursor (SOP)) cells. Our evidence suggests that caspase regulation of the wingless pathway is not associated with apoptotic cell death. These results imply a novel role for caspases in modulating cell signaling pathways through substrate cleavage in neural precursor development. © 2005 European Molecular Biology Organization

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Essential role of Shp2-binding sites on FRS2α for corticogenesis and for FGF2-dependent proliferation of neural progenitor cells

Yamamoto S., Yoshino I., Shimazaki T., Murohashi M., Hevner R. F., Lax I., Okano H., Shibuya M., Schlessinger J., Gotoh N.

Proceedings of the National Academy of Sciences of the United States of America 102 （ 44 ） 15983 - 15988 2005年11月

ISSN 0027-8424

Mammalian corticogenesis occurs through a complex process that includes neurogenesis, in which neural progenitor cells proliferate, differentiate, and migrate. It has been reported recently that neurogenesis occurs in the subventricular zone (SVZ), a region previously thought to be the primary site of gliogenesis. It has been recognized that in the SVZ, intermediate progenitor cells, derived from radial glial cells that are multipotent neural stem cells, produce only neurons. However, the molecular mechanisms underlying the regulation of neural stem cells and intermediate progenitor cells as well as their contribution to overall corticogenesis remain unknown. The docking protein FRS2α is a major mediator of signaling by means of FGFs and neurotrophins. FRS2α mediates many of its pleiotropic cellular responses by recruiting the adaptor protein Grb2 and the protein tyrosine phosphatase Shp2 upon ligand stimulation. Here, we report that targeted disruption of Shp2-binding sites in FRS2α leads to severe impairment in cerebral cortex development in mutant mice. The defect in corticogenesis appears to be due at least in part to abnormalities in intermediate progenitor cells. Genetic evidence is provided that FRS2α plays critical roles in the maintenance of intermediate progenitor cells and in neurogenesis in the cerebral cortex. Moreover, FGF2-responsive neurospheres, which are cell aggregates derived from neural stem/progenitor cells (NSPCs), from FRS2α mutant mice were smaller than those of WT mice. However, mutant NSPCs were able to self-renew, demonstrating that Shp2-binding sites on FRS2α play an important role in NSPC proliferation but are dispensable for NSPC self-renewing capacity after FGF2 stimulation. © 2005 by The National Academy of Sciences of the USA.

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Transplantation of neural stem cells for spinal cord injury

Nakamura Masaya, Toyama Yoshiaki, Okano Hideyuki

Clinical Neurology 45 （ 11 ） 874 - 876 2005年11月

ISSN 0009-918X

Neural progenitor cells, including neural stem cells (NSCs), are an important potential graft material for cell therapeutics of damaged spinal cord. Here we used as a source of graft material a NSC-enriched population derived from human fetal spinal cord (Embryonic week 8-9) and expanded in vitro by neurosphere formation. NSCs labeled with BrdU (TP) or culture medium (CON) were transplanted into the adult marmoset spinal cord after contusion injury at C5 level. Grafted NSCs survived and migrated up to 7 mm far from the lesion epicenter. Double-staining with TuJ1 for neuron, GFAP for astrocyte, or CNPase for oligodendrocyte and BrdU revealed that grafted NSCs differentiated into neurons and oligodendrocytes 8 weeks after transplantation. More neurofilaments were observed in TP than those of CON. Furthermore, behavioral assessment of forelimb muscle strength using bar grip test and amount of spontaneous motor activity using infrared-rays monitoring revealed that the grafted NSCs significantly increased both of them compared to those of CON. These results indicate that in vitro expanded NSCs derived from human fetal spinal cord are useful sources for the therapeutics of spinal cord injury in primates.

An epidural cooling catheter protects the spinal cord against ischemic injury in pigs

Mori Atsuo, Ueda Toshihiko, Hachiya Takashi, Kabei Nobuyuki, Okano Hideyuki, Yozu Ryohei, Sasaki Tatsuumi

Annals of Thoracic Surgery 80 （ 5 ） 1829 - 1833 2005年11月

ISSN 0003-4975

Background. Using swine, we investigated whether epidural placement of a cooling catheter rather than infusing iced saline solution could protect the spinal cord from ischemia during aortic surgery. Methods. We divided 14 domestic pigs into two groups of 7 each. Each underwent epidural catheter placement preceding 30 minutes of aortic cross-clamping distal to the origin of the left subclavian artery. In group 1, cold water was circulated continuously through the lumen of the catheter connected to an external unit. In group 2, animals received catheter placement without cooling. Spinal cord somatosensory evoked potentials were recorded. Neurologic status involving hind limbs was graded sequentially after surgery. Results. At aortic cross-clamping, spinal temperature in group 1 (31.7° ± 0.6°C) was significantly lower than in group 2 (37.8° ± 0.4°C; p < 0.0001). No significant elevation of intrathecal pressure accompanied cooling with the catheter (group 1, 8.1 ± 1.7 mm Hg; group 2, 8.0 ± 1.5 mm Hg). Mean duration of total loss of potentials was significantly shorter in group 1 (7.4 ± 3.8 minutes) than group 2 (19.7 ± 7.3 minutes; p = 0.0002). Pigs in group 1 exhibited better hind limb function recovery (mean Tarlov score, 4.7 ± 0.5) than group 2 (0.6 ± 0.8; p = 0.0017). Group 1 showed normal histologic characteristics, whereas group 2 showed loss of motor neurons in the ventral horns. Conclusions. Epidural cooling catheter without iced saline infusion can cool the spinal cord without elevating intrathecal pressure, protecting the cord against ischemia. © 2005 by The Society of Thoracic Surgeons.

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Identification of neural stem cells in adult human brain

Okano Hideyuki

Clinical Neurology 45 （ 11 ） 871 - 873 2005年11月

ISSN 0009-918X

Neural stem cells (NSCs) ar self-renewing, multipotential progenitor cells. A single NSC can give rise to a wide variety of CNS cells, including neurons, astrocytes, and oligodendrocytes. Because of these characteristics, there is an increasing interest in NSCs and neural progenitor cells, both from a basic developmental biology perspective and from a clinical one that is aimed at developing therapeutic applications for the damaged brain. Current research into the nature of the NSCs present in the CNS includes the study of the extracellular factors and signal transduction cascades involved in their differentiation and maintenance, their population dynamics, and their localization in the embryonic and adult brain. These lines of research, combined with other studies intended to permit the prospective identification and isolation of NSCs, and their induction into particular neuronal phenotypes-which will be introduced in my talk-should lead to the development of feasible strategies for manipulating NSC cells in situ to treat the damaged brain and spinal cord injury.

In vivo imaging of engrafted neural stem cells

Okada Seiji, Ishii Ken, Yamane Jinichi, Iwanami Akio, Ikegami Takeshi, Katoh Hiroyuki, Iwamoto Yukihide, Nakamura Masaya, Miyoshi Hiroyuki, Okano Hirotaka James, Contag Christopher H., Toyama Yoshiaki, Okano Hideyuki

FASEB Journal 19 （ 13 ） 1839 - 1841 2005年11月

ISSN 0892-6638

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Neural stem cells and regeneration of injured spinal cord

Okano Hideyuki, Okada Seiji, Nakamura Masaya, Toyama Yoshiaki

Kidney International 68 （ 5 ） 1927 - 1931 2005年11月

ISSN 0085-2538

Recent progress in the stem cell biology has led much insight into new therapeutic interventions aiming for the regeneration of the damaged central nervous system. The major strategies can be classified into two subgroups: (1) activation of endogenous neural stem cells, and (2) cell transplantation therapies. In either of these strategies, it is crucial to understand the underlying mechanisms of maintenance, activation, and differentiation of neural stem cells and subsequent process, including the migration, survival, and functional maturation of differentiated cells. In this paper, we would like to summarize our recent findings on the therapeutic interventions of the injured spinal cord, especially focusing on the development of treatment for the acute phase of spinal cord injury with anti-interleukin (IL)-6 receptor blocking antibody. © 2005 by the International Society of Nephrology.

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Nedd9 protein, a Cas-L homologue, is upregulated after transient global ischemia in rats

Sasaki Takahiro, Iwata Satoshi, Okano Hirotaka James, Urasaki Yasuyo, Hamada Junichi, Tanaka Hirotoshi, Dang Nam H., Okano Hideyuki, Morimoto Chikao

Stroke 36 （ 11 ） 2457 - 2462 2005年11月

ISSN 0039-2499

Background and Purpose - Some proteins involved in self-repair after stroke in the adult brain are primarily expressed during embryonic development and strongly down-regulated during the early postnatal phase. Neuronal precursor cell-expressed, developmentally down-regulated gene (Nedd) 9 was recognized to be identical to Crk-associated substrate lymphocyte type (Cas-L), a docking protein that associates with a variety of signaling molecules, such as focal adhesion kinase (FAK), proline-rich tyrosine kinase 2 (Pyk2), and Crk. We investigated the involvement of these proteins in the pathophysiology of global cerebral ischemia. Methods - The mouse Cas-L/Nedd9 cDNAs were cloned. The expression and function of Cas-L/Nedd9 protein in the pathogenesis of global ischemia in rats was investigated by RT-PCR, Western blot analysis, and immunohistochemistry. The neurite outgrowth of the transfectants of Nedd9 deletion mutants in PC-12 cells was also assessed to clarify the function of the Nedd9 protein. Results - Nedd9 was a splicing variant of Cas-L and was selectively induced in neurons of the cerebral cortex and hippocampus 1 to 14 days after the ischemia. Induced Nedd9 protein was tyrosine phosphorylated and was bound to FAK in dendrite and soma of neurons after the ischemia. Finally, it was demonstrated that Nedd9 promoted neurite outgrowth of PC-12 cells. Conclusions - Our study may support the potential of Nedd9 for participation in the differentiation of neurons after global ischemia in rats. © 2005 American Heart Association, Inc.

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Visualization of spatiotemporal activation of Notch signaling

Kohyama Jun, Tokunaga Akinori, Fujita Yuko, Miyoshi Hiroyuki, Nagai Takeharu, Miyawaki Atsushi, Nakao Keiko, Matsuzaki Yumi, Okano Hideyuki

Developmental Biology 286 （ 1 ） 311 - 325 2005年10月

ISSN 0012-1606

Notch signaling plays various key roles in cell fate determination during CNS development in a context-dependent fashion. However, its precise physiological role and the localization of its target cells remain unclear. To address this issue, we developed a new reporter system for assessing the RBP-J-mediated activation of Notch signaling target genes in living cells and tissues using a fluorescent protein Venus. Our reporter system revealed that Notch signaling is selectively activated in neurosphere-initiating multipotent neural stem cells in vitro and in radial glia in the embryonic forebrain in vivo. Furthermore, the activation of Notch signaling occurs during gliogenesis and is required in the early stage of astroglial development. Consistent with these findings, the persistent activation of Notch signaling inhibits the differentiation of GFAP-positive astrocytes. Thus, the development of our RBP-J-dependent live reporter system, which is activated upon Notch activation, together with a stage-dependent gain-of-function analysis allowed us to gain further insight into the complexity of Notch signaling in mammalian CNS development. © 2005 Elsevier Inc. All rights reserved.

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Expression of a candidate marker for progenitor cells, Musashi-1, in the proliferative regions of human antrum and its decreased expression in intestinal metaplasia

Akasaka Yoshikiyo, Saikawa Y., Fujita K., Kubota T., Ishikawa Y., Fujimoto A., Ishii T., Okano H., Kitajima M.

Histopathology 47 （ 4 ） 348 - 356 2005年10月

ISSN 0309-0167

Aim: Reliable makers for progenitor cells in the human stomach have not been elucidated. The aim of the present study was to clarify whether Musashi-1 (Msi-1), which has recently been proposed as a stem cell marker in mouse intestine, serves as a marker for progenitor cells in human stomach. Methods and results: Immunohistochemistry revealed that Msi-1+ cells were detected especially in the isthmus/neck region (the putative position of stem cells) of the adult antrum, but were limited to the basal regions of fetal pyloric glands during the early stages of development. These results suggest that Msi-1 expression occurs specifically in the stem cell-containing regions. Msi-1+ cells were intermingled with proliferating cell nuclear antigen (PCNA)+ cells in the isthmus/neck region of the adult antrum, but did not coexpress PCNA or Ki67. Msi-1 expression overlapped partly with expression of MUC5AC and MUC6, indicating that Msi-1+ cells retain some features of both foveolar and pyloric gland cell differentiation phenotypes. In contrast, Msi-1 expression in gastric glands showing intestinal metaplasia (IM) became weaker than that in the glands without IM. Conclusion: The specific expression of Msi-1 within the proliferative regions suggests that Msi-1 is a marker of cells with progenitor characteristics before active proliferation in human antrum. © 2005 Blackwell Publishing Limited.

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Correlation between Musashi-1 and c-hairy-1 expression and cell proliferation activity in the developing intestine and stomach of both chicken and mouse

Asai Rieko, Okano Hideyuki, Yasugi Sadao

Development Growth and Differentiation 47 （ 8 ） 501 - 510 2005年10月

ISSN 0012-1592

Musashi-1 (Msi-1) is an RNA-binding protein that plays key roles in the maintenance of neural stem cell states and in their differentiation into neural cells. Msi-1 has also been proposed as a candidate marker gene of mammalian intestinal stem cells and their immediate lineages. In this study, we examined Msi-1 expression in the small intestine and the stomach of both chicken and mouse during embryonic, fetal and postnatal development. In addition, we analyzed the expression of c-hairy-1, a chicken homologue of mouse Hes1, and assessed the proliferative activity of the cells expressing both of these factors. Significantly, during the development of these digestive organs in both species Msi-1 expression showed dynamic changes, suggesting that it is important for digestive organ development, particularly for epithelial differentiation. Based on our observations of the expression patterns of Msi-1 and c-hairy-1 in the adult small intestine, we speculate that Msi-1 is also a stem cell marker of the chicken small intestinal epithelium.

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Establishment of novel embryonic stem cell lines derived from the common marmoset (Callithrix jacchus)

Sasaki Erika, Hanazawa Kisaburo, Kurita Ryo, Akatsuka Akira, Yoshizaki Takahito, Ishii Hajime, Tanioka Yoshikuni, Ohnishi Yasuyuki, Suemizu Hiroshi, Sugawara Ayako, Tamaoki Norikazu, Izawa Kiyoko, Nakazaki Yukoh, Hamada Hiromi, Suemori Hirofumi, Asano Shigetaka, Nakatsuji Norio, Okano Hideyuki, Tani Kenzaburo

Stem Cells 23 （ 9 ） 1304 - 1313 2005年10月

ISSN 1066-5099

The successful establishment of human embryonic stem cell (hESC) lines has inaugurated a new era in regenerative medicine by facilitating the transplantation of differentiated ESCs to specific organs. However, problems with the safety and efficacy of hESC therapy in vivo remain to be resolved. Preclinical studies using animal model systems, including nonhuman primates, are essential to evaluate the safety and efficacy of hESC therapies. Previously, we demonstrated that common marmosets are suitable laboratory animal models for preclinical studies of hematopoietic stem cell therapies. As this animal model is also applicable to preclinical trials of ESC therapies, we have established novel common marmoset ESC (CMESC) lines. To obtain marmoset embryos, we developed a new embryo collection system, in which blastocysts can be obtained every 3 weeks from each marmoset pair. The inner cell mass was isolated by immunosurgery and plated on a mouse embryonic feeder layer. Some of the CMESC lines were cultured continuously for more than 1 year. These CMESC lines showed alkaline phosphatase activity and expressed stage-specific embryonic antigen (SSEA)-S, SSEA-4, TRA-1-60, and TRA-1-81. On the other hand, SSEA-1 was notdetected. Furthermore, our novel CMESCs are pluripotent, as evidenced by in vivo teratoma formation in immunodeficient mice and in vitro differentiation experiments. Our established CMESC lines and the common marmoset provide an excellent experimental model system for understanding differentiation mechanisms, as well as the development of regenerative therapies using hESCs. ©Alpha Med Press.

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Cardiac neural crest cells contribute to the dormant multipotent stem cell in the mammalian heart

Tomita Yuichi, Matsumura Keisuke, Wakamatsu Yoshio, Matsuzaki Yumi, Shibuya Isao, Kawaguchi Haruko, Ieda Masaki, Kanakubo Sachiko, Shimazaki Takuya, Ogawa Satoshi, Osumi Noriko, Okano Hideyuki, Fukuda Keiichi

Journal of Cell Biology 170 （ 7 ） 1135 - 1146 2005年09月

ISSN 0021-9525

Arodent cardiac side population cell fraction formed clonal spheroids in serum-free medium, which expressed nestin, Musashi-1, and multidrug resistance transporter gene 1, markers of undifferentiated neural precursor cells. These markers were lost following differentiation, and were replaced by the expression of neuron-, glial-, smooth muscle cell-, or cardiomyocyte-specific proteins. Cardiosphere-derived cells transplanted into chick embryos migrated to the truncus arteriosus and cardiac outflow tract and contributed to dorsal root ganglia, spinal nerves, and aortic smooth muscle cells. Lineage studies using double transgenic mice encoding protein 0-Cre/Floxed-EGFP revealed undifferentiated and differentiated neural crest-derived cells in the fetal myocardium. Undifferentiated cells expressed GATA-binding protein 4 and nestin, but not actinin, whereas the differentiated cells were identified as cardiomyocytes. These results suggest that cardiac neural crest-derived cells migrate into the heart, remain there as dormant multipotent stem cells-and under the right conditions-differentiate into cardiomyocytes and typical neural crest-derived cells, including neurons, glia, and smooth muscle. © The Rockefeller University Press.

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Enhanced proliferation of progenitor cells in the subventricular zone and limited neuronal production in the striatum and neocortex of adult macaque monkeys after global cerebral ischemia

Tonchev Anton B., Yamashima Tetsumori, Sawamoto Kazunobu, Okano Hideyuki

Journal of Neuroscience Research 81 （ 6 ） 776 - 788 2005年09月

ISSN 0360-4012

Cerebral ischemia in adult rodent models increases the proliferation of endogenous neural progenitor cells residing in the subventricular zone along the anterior horn of the lateral ventricle (SVZa) and induces neurogenesis in the postischemic striatum and cortex. Whether the adult primate brain preserves a similar ability in response to an ischemic insult is uncertain. We used the DNA synthesis indicator bromodeoxyuridine (BrdU) to label newly generated cells in adult macaque monkeys and show here that the proliferation of cells with a progenitor phenotype (double positive for BrdU and the markers Musashi1, Nestin, and βIII-tubulin) in SVZa increased during the second week after a 20-min transient global brain ischemia. Subsequent progenitor migration seemed restricted to the rostral migratory stream toward the olfactory bulb and ischemia increased the proportion of adult-generated cells retaining their location in SVZa with a progenitor phenotype. Despite the lack of evidence for progenitor cell migration toward the postischemic striatum or prefrontal neocortex, a small but sustained proportion of BrdU-labeled cells expressed features of postmitotic neurons (positive for the protein NeuN and the transcription factors Tbr1 and Islet1) in these two regions for at least 79 days after ischemia. Taken together, our data suggest an enhanced neurogenic response in the adult primate telencephalon after a cerebral ischemic insult. © 2005 Wiley-Liss, Inc.

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Inhibition of glioma cell proliferation by neural stem cell factor

Suzuki Tsuyoshi, Izumoto Shuichi, Wada Kouichi, Fujimoto Yasunori, Maruno Motohiko, Yamasaki Mami, Kanemura Yonehiro, Shimazaki Takuya, Okano Hideyuki, Yoshimine Toshiki

Journal of Neuro-Oncology 74 （ 3 ） 233 - 239 2005年09月

ISSN 0167-594X

Neural stem cells (NSC) have unique differentiation-, proliferation-, and motility properties. To investigate whether they secrete factors that interfere with the proliferation of glioma cells, we grew glioma cells in conditioned medium (CM) obtained from cultures of neurospheres including neural stem/progenitor cells (NSPC) isolated from embryonic (E14)- or adult mouse brain or fetal human brain. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and BrdU-labeling assays showed that CM from NSPC (NSPC/CM) contained factor(s) that inhibited the proliferation of glioma cells by 28-87%. Filter-fractionation of NSPC/CM revealed that the 50,000-100,000 nominal molecular weight limit (NMWL) fraction contained the inhibitory activity. On the basis of these observations we transplanted 203G glioma cells and/or NSPC into the intrathecal space of the cisterna magna of mice to investigate whether NSPC interfere with the proliferation of glioma cells in vivo. Mice transplanted with both 203G and NSPC survived significantly longer than did mice transplanted only with 203G. We concluded that NSPC secrete factor(s) that may control glioma cell proliferation. © Springer 2005.

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Targeted expression of anti-apoptotic protein p35 in oligodendrocytes reduces delayed demyelination and functional impairment after spinal cord injury

Tamura Mutsuhiro, Nakamura Masaya, Ogawa Yuto, Toyama Yoshiaki, Miura Masayuki, Okano Hideyuki

GLIA 51 （ 4 ） 312 - 321 2005年09月

ISSN 0894-1491

Functional impairment after spinal cord injury (SCI) is attributed to neuronal cell necrosis death and axonotmesis, with further worsening caused by the accompanying apoptosis of myelin-forming oligodendrocytes (OLGs). However, it is unclear as to how much OLG apoptosis contributes to functional impairment. To address this issue, we used transgenic mice characterized by the targeted expression of p35, a broad-spectrum caspase inhibitor, in OLGs using the cre/loxP system (referred to as cre/p35 transgenic mice). In this study, we examined the motor function and histopathologic changes after a contusive thoracic spinal cord injury in the cre/p35 transgenic mice. A larger number of OLGs and a lesser extent of demyelination were observed after SCI in the cre/p35 transgenic mice than in the control cre mice, which did not carry the p35 transgene. Furthermore, the motor function of the hindlimbs recovered to a significantly better degree in the cre/p35 transgenic mice than in the control ere mice. Thus, the inhibition of OLG apoptosis decreased the extent of functional impairment after SCI. These findings suggest that the inhibition of OLG apoptosis may be a potential treatment for SCI. ©2005 Wiley-Liss, Inc.

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Transplantation of embryonic spinal cord-derived neurospheres support growth of supraspinal projections and functional recovery after spinal cord injury in the neonatal rat

Nakamura M., Okano H., Toyama Y., Dai H. N., Finn T. P., Bregman Barbara S.

Journal of Neuroscience Research 81 （ 4 ） 457 - 468 2005年08月

ISSN 0360-4012

Great interest exists in using cell replacement strategies to repair the damaged central nervous system. Previous studies have shown that grafting rat fetal spinal cord into neonate or adult animals after spinal cord injury leads to improved anatomic growth/plasticity and functional recovery. It is clear that fetal tissue transplants serve as a scaffold for host axon growth. In addition, embryonic Day 14 (E14) spinal cord tissue transplants are also a rich source of neural-restricted and glial-restricted progenitors. To evaluate the potential of E14 spinal cord progenitor cells, we used in vitro-expanded neurospheres derived from embryonic rat spinal cord and showed that these cells grafted into lesioned neonatal rat spinal cord can survive, migrate, and differentiate into neurons and oligodendrocytes, but rarely into astrocytes. Synapses and partially myelinated axons were detected within the transplant lesion area. Transplanted progenitor cells resulted in increased plasticity or regeneration of corticospinal and brainstem-spinal fibers as determined by anterograde and retrograde labeling. Furthermore, transplantation of these cells promoted functional recovery of locomotion and reflex responses. These data demonstrate that progenitor cells when transplanted into neonates can function in a similar capacity as transplants of solid fetal spinal cord tissue. © 2005 Wiley-Liss, Inc.

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Comparison of various bone marrow fractions in the ability to participate in vascular remodeling after mechanical injury

Sahara Makoto, Sata Masataka, Matsuzaki Yumi, Tanaka Kimie, Morita Toshihiro, Hirata Yasunobu, Okano Hideyuki, Nagai Ryozo

Stem Cells 23 （ 7 ） 874 - 878 2005年08月

ISSN 1066-5099

In contrast to conventional assumption, recent reports propose the possibility that hematopoietic stem cells (HSCs) may have broader potential to differentiate into various cell types. Here, we tested the pluripotency of HSCs by comparing vascular lesions induced by mechanical injury after bone marrow reconstitution with total bone marrow (TBM) cells, C-KH⁺ Sca-1 ⁺ Lin- (KSL) cells, or a single HSC cell (Tip-SP CD34-KSL cell, CD34- c-Kit⁺ Sca-1⁺ Lin- cell with the strongest dyeefflux activity) harboring green fluorescent protein (GFP). The lesions contained a significant number of GFP-positive cells in the TBM and KSL groups, whereas GFP-positive cells were rarely detected in the HSC group. These results suggest that transdifferentiation of a highly purified HSC seems to be a rare event, if it occurs at all, whereas bone marrow cells including the KSL fraction can give rise to vascular cells that substantially contribute to repair or lesion formation after mechanical injury. ©AlphaMed Press.

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Inflammation and regeneration of injured spinal cord: effects of the stem cell transplantation and blockade of IL-6 signaling

岡野栄之

INFLAMMATION RESEARCH 54 S92 - S92 2005年08月

研究論文（学術雑誌）, ISSN 1023-3830

Increased number of neural progenitors in human temporal lobe epilepsy

Crespel Arielle, Rigau Valérie, Coubes Philippe, Rousset Marie Claude, De Bock Frédéric, Okano Hideyuki, Baldy-Moulinier Michel, Bockaert Joël, Lerner-Natoli Mireille

Neurobiology of Disease 19 （ 3 ） 436 - 450 2005年08月

ISSN 0969-9961

An increased neurogenesis is reported in animal models of mesial temporal lobe epilepsy (MTLE) but the fate of newborn cells is unknown. Here, we attempted to demonstrate neurogenesis in adult epileptic tissue obtained after hippocampectomy. MTLE hippocampi showed increased expression of division markers and of Musashi-1, a marker of neural progenitors, compared to control hippocampi. Large quantities of Musashi-1⁺ cells were obvious in the subgranular layer and the subventricular zone, both known neurogenic areas, and in the fissura hippocampi. Musashi-1 was expressed by small cells that were mainly vimentin⁺ or nestin⁺, rarely Dcx⁺ or PSA-NCAM⁺ and negative for markers of mature neurons or astrocytes. Some of them are present in the granular layer, the hilus, and CA1 area resembling the ectopic positions described in rodents. These findings demonstrate that neural progenitors proliferate in chronic epilepsy and suggest that the fissura hippocampi behaves like another neurogenic area. © 2005 Elsevier Inc. All rights reserved.

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Characterization of ABC transporter ABCB1 expressed in human neural stem/progenitor cells

Islam Mohammed Omedul, Kanemura Yonehiro, Tajria Jesmin, Mori Hideki, Kobayashi Satoshi, Shofuda Tomoko, Miyake Jun, Hara Masayuki, Yamasaki Mami, Okano Hideyuki

FEBS Letters 579 （ 17 ） 3473 - 3480 2005年07月

ISSN 0014-5793

We investigated the localization and functional expression of the ABC transporter ABCB1 in human fetal neural stem/progenitor cells (hNSPCs). RT-PCR analysis revealed ABCB1 gene expression in hNSPCs. We found a single band in immunoblotted hNSPCs lysates probed with ABCB1 antibody, and detected ABCB1 at the hNSPCs cell membrane by immunocytochemistry and subcellular fractionation. ABCB1 inhibitors and substrate, and ATP-depleting agents enhanced hNSPCs' rhodamine 123 accumulation, and hNSPCs microsomes had vanadate-sensitive ATPase activity. ABCB1 and nestin expression decreased during hNSPCs differentiation, while the astroglial marker GFAP increased. ABCB1 may maintain hNSPCs in an undifferentiated state and could be a neural stem/progenitor marker. © 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

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Use of TH-EGFP transgenic mice as a source of identified dopaminergic neurons for physiological studies in postnatal cell culture

Jomphe C., Bourque M. J., Fortin G. D., St-Gelais F., Okano H., Kobayashi K., Trudeau L. E.

Journal of Neuroscience Methods 146 （ 1 ） 1 - 12 2005年07月

ISSN 0165-0270

The physiological and pharmacological properties of dopaminergic neurons in the brain are of major interest. Although much has been learned from cell culture studies, the physiological properties of these neurons remain difficult to study in such models because they are usually in minority and are difficult to distinguish from other non-dopaminergic neurons. Here we have taken advantage of a recently engineered transgenic mouse model expressing enhanced green fluorescence protein (EGFP) under the control of the tyrosine hydroxylase promoter to establish a more effective dopaminergic neuron cell culture model. We first evaluated the specificity of the EGFP expression. Although ectopic expression of EGFP was found in cultures derived from postnatal day 0 pups, this decreased over time in culture such that after 2 weeks, approximately 70% of EGFP-expressing neurons were dopaminergic. We next sought to validate this dopaminergic neuron culture model. We evaluated whether EGFP-expressing dopaminergic neurons displayed some of the well-established properties of dopaminergic neurons. Autoreceptor stimulation inhibited the activity of dopaminergic neurons while neurotensin receptor activation produced the opposite effect. Confocal imaging of the synaptic vesicle optical tracer FM4-64 in EGFP-expressing dopaminergic neurons demonstrated the feasibility of high resolution monitoring of the activity of single terminals established by these neurons. Together, this work provides evidence that primary cultures of postnatal TH-EGFP mice currently represent an excellent model to study the properties of these cells in culture. © 2005 Elsevier B.V. All rights reserved.

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Blockade of interleukin-6 signaling aggravates ischemic cerebral damage in mice

Yamashita Toru, Sawamoto Kazunobu, Suzuki Shigeaki, Suzuki Norihiro, Adachi Kazuhide, Kawase Takeshi, Mihara Masahiko, Ohsugi Yoshiyuki, Abet Koji, Okano Hideyuki

Journal of Neurochemistry 94 （ 2 ） 459 - 468 2005年07月

ISSN 0022-3042

Interleukin (IL)-6 expression transiently increases in the acute phase of cerebral ischemia. To investigate the physiological significance of endogenous IL-6 expression and to identify the main signal pathway for the action of IL-6, we administered anti-mouse IL-6 receptor monoclonal antibody (IL-6RA), which blocks IL-6 signaling, to mice immediately after a 45-min period of middle cerebral artery occlusion (MCAO). At 6 h after MCAO, IL-6RA administration had resulted in a significant reduction in the amount of phosphorylated signal transducer and activator of transcription-3 (Stat3) protein in the peri-infarct area of the cortex. At 24 h after MCAO, blockade of IL-6 signaling had led to an increase in number of apoptotic cells in the peri-infarct area and enlargement of the size of the infarct, and it had adversely affected neurological function. These results suggest that endogenous IL-6 plays a critical role in preventing damaged neurons from undergoing apoptosis in the acute phase of cerebral ischemia and that its role may be mediated by Stat3 activation. © 2005 International Society for Neurochemistry.

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Possibility for neurogenesis in substantia nigra of parkinsonian brain

Yoshimi Kenji, Ren Yong Ri, Seki Tatsunori, Yamada Masanori, Ooizumi Hideki, Onodera Masafumi, Saito Yuko, Murayama Shigeo, Okano Hideyuki, Mizuno Yoshikuni, Mochizuki Hideki

Annals of Neurology 58 （ 1 ） 31 - 40 2005年07月

ISSN 0364-5134

Recent studies of enhanced hippocampal neurogenesis by antidepressants suggest enhancement of neurogenesis is a potentially effective therapy in neurodegenerative diseases. In this study, we evaluated nigral neurogenesis in animals and autopsy brains including patients with Parkinson's disease (PD). First, proliferating cells in substantia nigra were labeled with retroviral transduction of green fluorescent protein, which is an efficient method to label neuronal stem cells. Subsequent differentiation of labeled cells was followed; many transduced cells became microglia, but no differentiation into tyrosine hydroxylase-positive neurons was detected at 4 weeks after injection, in both intact rodents and those treated with 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine. Second, polysialic acid (PSA)-like immunoreactivity, indicative of newly differentiated neurons, was detected in the substantia nigra of rodent, primate, and human midbrains. A large number of PSA-positive cells were detected in the substantia nigra pars reticulata of some patients with PD. In rats and a macaque monkey, the dopamine-depleted hemispheres showed more PSA staining than the intact side. A small number of tyrosine hydroxylase-positive cells were PSA-positive. Our results suggest enhanced neural reconstruction in PD, which may be important in the design of new therapies against the progression of PD. © 2005 American Neurological Association.

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Potential functional neural repair with grafted neural stem cells of early embryonic neuroepithelial origin

Uchida Koichi, Momiyama Toshihiko, Okano Hideyuki, Yuzaki Michisuke, Koizumi Amane, Mine Yutaka, Kawase Takeshi

Neuroscience Research 52 （ 3 ） 276 - 286 2005年07月

ISSN 0168-0102

The fate of grafted neuroepithelial stem cells in the normal mature brain environment was assessed both morphologically and electrophysiologically to confirm their feasibility in the functional repair of damaged neural circuitry. The neuroepithelial stem cells were harvested from the mesencephalic neural plate of transgenic green fluorescence protein-carrying rat embryos, and implanted into the normal adult rat striatum. The short- and long-term differentiation pattern of donor-derived cells was precisely monitored immunohistochemically. The functional abilities of the donor-derived cells and communication between them and the host were investigated using host-rat brain slices incorporating the graft with whole-cell patch-clamp recording. Vigorous differentiation of the neuroepithelial stem cells into mostly neurons was noted in the short-term with positive staining for tyrosine hydroxylase, suggesting that the donor-derived cells were exclusively following their genetically programmed fate, together with gamma-aminobutyric acid (GABA) and glutamate expression. In the long-term, the large number of donor-derived neurons was sustained, but the staining pattern showed expression of dopamine- and adenosine 3′:5′-monophosphate-regulated phosphoprotein 32, suggesting that some neurons were following environmental cues, together with the appearance of some cholinergic neurons. Some donor-derived astrocytes were also seen in the graft. Many action potentials indicating the presence of both dopaminergic and non-dopaminergic patterns could be elicited and recorded in the donor-derived neurons in addition to spontaneous glutamatergic and GABAergic post-synaptic currents which were strongly shown to be of host origin. Neuroepithelial stem cells are therefore an attractive candidate as a source of donor material for intracerebral grafting in functional repair. © 2005 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

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Function of RNA-binding protein Musashi-1 in stem cells

Okano Hideyuki, Kawahara Hironori, Toriya Masako, Nakao Keio, Shibata Shinsuke, Imai Takao

Experimental Cell Research 306 （ 2 ） 349 - 356 2005年06月

ISSN 0014-4827

Musashi is an evolutionarily conserved family of RNA-binding proteins that is preferentially expressed in the nervous system. The first member of the Musashi family was identified in Drosophila. This protein plays an essential role in regulating the asymmetric cell division of ectodermal precursor cells known as sensory organ precursor cells through the translational regulation of target mRNA. In the CNS of Drosophila larvae, however, Musashi is expressed in proliferating neuroblasts and likely has a different function. Its probable mammalian homologue, Musashi-1, is a neural RNA-binding protein that is strongly expressed in fetal and adult neural stem cells (NSCs). Mammalian Musashi-1 augments Notch signaling through the translational repression of its target mRNA, m-Numb, thereby contributing to the self-renewal of NSCs. In addition to its functions in NSCs, the role of mammalian Musashi-1 protein in epithelial stem cells, including intestinal and mammary gland stem cells, is attracting increasing interest. © 2005 Elsevier Inc. All rights reserved.

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Musculin/MyoR is expressed in kidney side population cells and can regulate their function

Hishikawa Keiichi, Marumo Takeshi, Miura Shigeki, Nakanishi Asato, Matsuzaki Yumi, Shibata Katsunori, Ichiyanagi Tomoko, Kohike Hiroko, Komori Takuya, Takahashi Ichiro, Takase Osamu, Imai Naohiko, Yoshikawa Masahiro, Inowa Toshihiko, Hayashi Matsuhiko, Nakaki Toshio, Nakauchi Hiromitsu, Okano Hideyuki, Fujita Toshiro

Journal of Cell Biology 169 （ 6 ） 921 - 928 2005年06月

ISSN 0021-9525

Musculin/MyoR is a new member of basic helixloop-helix transcription factors, and its expression is limited to skeletal muscle precursors. Here, we report that musculin/MyoR is expressed in adult kidney side population (SP) cells and can regulate their function. SP phenotype can be used to purify stem cell-rich fractions. Microarray analysis clarified that musculin/MyoR was exclusively expressed in kidney SP cells, and the cells resided in the renal interstitial space. Musculin/MyoR-positive cells were decreased in acute renal failure, but infusion of kidney SP cells increased musculin/MyoR-positive cells and improved renal function. Kidney SP cells in reversible acute renal failure expressed a high level of renoprotective factors and leukemia inhibitory factor (LIF), but not in irreversible chronic renal failure. In cultured kidney SP cells, LIF stimulated gene expression of renoprotective factors, and down-regulation of musculin/MyoR augmented LIF-induced gene expression. Our results suggest that musculin/MyoR may play important roles not only in developmental processes but also in regenerative processes in adult tissue. © The Rockefeller University Press.

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Drosophila deltex mediates suppressor of hairless-independent and late-endosomal activation of notch signaling

岡野栄之

CELL STRUCTURE AND FUNCTION 30 68 - 68 2005年06月

研究論文（学術雑誌）, ISSN 0386-7196

Role of autophagy in protein quality control in the central nervous system

岡野栄之

CELL STRUCTURE AND FUNCTION 30 49 - 49 2005年06月

研究論文（学術雑誌）, ISSN 0386-7196

Increase of bone marrow-derived secretory lineage epithelial cells during regeneration in the human intestine

Matsumoto Tomoko, Okamoto Ryuichi, Yajima Tomoharu, Mori Takehiko, Okamoto Shinichiro, Ikeda Yasuo, Mukai Makio, Yamazaki Motomi, Oshima Shigeru, Tsuchiya Kiichiro, Nakamura Tetsuya, Kanai Takanori, Okano Hideyuki, Inazawa Johji, Hibi Toshifumi, Watanabe Mamoru

Gastroenterology 128 （ 7 ） 1851 - 1867 2005年06月

ISSN 0016-5085

Background & Aims: We have previously reported that bone marrow (BM)-derived cells contribute to the regeneration of the human intestinal epithelium. To analyze further how these cells arise, proliferate, and differentiate as epithelial cells, histologic analysis was conducted using endoscopic specimens. Methods: Thirty biopsy specimens from 14 female, sex-mismatched BM-transplantation recipients were examined. BM-derived cells were identified by fluorescent in situ hybridization (FISH) for the Y chromosome and immunohistochemistry. Multicolor FISH was used to exclude cell fusion. These cells were further analyzed for various differentiation or proliferation markers. Results: No evidence of cell fusion was detected. BM-derived cells did not distribute within the crypt as stem cells and rarely expressed Musashi-1. However, BM-derived epithelial cells frequently expressed Ki-67, and some of these cells appeared as pairs of adjacent cells. These cells also expressed markers of all 4 lineages of terminally differentiated cells. During regeneration following graft-vs-host disease, the number of BM-derived cells was substantially increased within Ki-67-positive cells. Interestingly, the number of cells expressing markers for secretory lineage cells was significantly increased within BM-derived cells. This change was unique for BM-derived cells, resulting in a significantly increased proportion of BM-derived cells among secretory lineage cells. Conclusions: BM-derived epithelial cells arise via a mechanism other than cell fusion and rarely give rise to stem cells. However, a small proportion of these cells express proliferation markers, and a majority reside as terminally differentiated cells. During regeneration BM-derived cells increase as secretory lineage cells, thereby contributing to restore epithelial functions. © 2005 by the American Gastroenterological Association.

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Notch signaling functions as a binary switch for the determination of glandular and luminal fates of endodermal epithelium during chicken stomach development

Matsuda Yoshimasa, Wakamatsu Yoshio, Kohyama Jun, Okano Hideyuki, Fukuda Kimiko, Yasugi Sadao

Development 132 （ 12 ） 2783 - 2793 2005年06月

ISSN 0950-1991

During development of the chicken proventriculus (glandular stomach), gut endoderm differentiates into glandular and luminal epithelium. We found that Delta1-expressing cells, undifferentiated cells and Notch-activated cells colocalize within the endodermal epithelium during early gland formation. Inhibition of Notch signaling using Numb or dominant-negative form of Su(H) resulted in a luminal differentiation, while forced activation of Notch signaling promoted the specification of immature glandular cells, but prevented the subsequent differentiation and the imagination of the glands. These results suggest that Delta1-mediated Notch signaling among endodermal cells functions as a binary switch for determination of glandular and luminal fates, and regulates patterned differentiation of glands in the chicken proventriculus.

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Role of IL-6 in spinal cord injury in a mouse model

Nakamura Masaya, Okada Seiji, Toyama Yoshiaki, Okano Hideyuki

Clinical Reviews in Allergy and Immunology 28 （ 3 ） 197 - 203 2005年06月

ISSN 1080-0549

In recent years, various studies have been conducted toward the goal of achieving regeneration of the central nervous system using neural stem cells. However, various complex factors are involved in the regulation of neural stem cell differentiation, and many unresolved questions remain. It has been reported that after spinal cord injury, the intrinsic neural stem cells do not differentiate into neurons but, rather, into astrocytes, resulting in the formation of glial scars. Based on reports that the expression of interleukin (IL)-6 and the IL-6 receptor (IL-6R) is sharply increased in the acute stages after spinal cord injury and that IL-6 may serve as a factor strongly inducing the differentiation of neural stem cells into astrocytes, we examined the effects of an antibody to IL-6R in cases of spinal cord injury and found that the antibody suppressed secondary injury (caused by inflammatory reactions) and glial scar formation, facilitating functional recovery. This article presents the data from this investigation and discusses the relationship between IL-6 signals and spinal cord injury. © Copyright 2005 by Humana Press Inc.

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Activation of cytokine signaling through leukemia inhibitory factor receptor (LIFR)/gp130 attenuates ischemic brain injury in rats

Suzuki Shigeaki, Yamashita Toru, Tanaka Kortaro, Hattori Hidenori, Sawamoto Kazunobu, Okano Hideyuki, Suzuki Norihiro

Journal of Cerebral Blood Flow and Metabolism 25 （ 6 ） 685 - 693 2005年06月

ISSN 0271-678X

Cytokine signaling through leukemia inhibitory factor receptor (LIFR)/gp130 is known to exert a neurotrophic action in the central nervous system, although the role of this signaling in cerebral ischemia remains unknown. We examined the effect of intracerebral injection of LIF after focal cerebral ischemia in rats. The animals underwent a sham operation (sham group) or middle cerebral artery occlusion (MCAO) followed by direct injection of either vehicle (phosphate-buffered saline, the PBS group) or recombinant LIF (10 ng in the low-LIF group and 100 ng in the high-LIF group) into the cerebral cortex adjacent to the inner boundary zone of the infarct area, and neurologic and histologic evaluations were conducted 24 h later. Expression of LIFR, gp130, and phosphorylated Stat3, Akt, and ERK1/2 was investigated by Western blot analysis and immunohistochemistry. The neurologic deficits and ischemic damage were significantly less severe in the high-LIF group than in the PBS group and the low-LIF group. Leukemia inhibitory factor receptor and gp130 were expressed in neurons, and the ischemic damage of these proteins was rescued in the high-LIF group. Early induction of phosphorylated Stat3 was significantly detected on the ischemic side in the high-LIF group after LIF injection. Exogenous LIF attenuates ischemic brain injury by activating cytokine signaling through LIFR/gp130. © 2005 ISCBFM All rights reserved.

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Prospective characterization of neural stem cells by flow cytometry analysis using a combination of surface markers

Nagato Masako, Heike Toshio, Kato Takeo, Yamanaka Yasunari, Yoshimoto Momoko, Shimazaki Takuya, Okano Hideyuki, Nakahata Tatsutoshi

Journal of Neuroscience Research 80 （ 4 ） 456 - 466 2005年05月

ISSN 0360-4012

Neural stem cells (NSCs) with self-renewal and multilineage differentiation properties can potentially repair degenerating or damaged neural tissue. Here, we have enriched NSCs from neurospheres, which make up a heterogeneous population, by fluorescence-activated cell sorting (FACS) with antibodies against syndecan-1, Notch-1, and integrin-β1, which were chosen as candidates for hematopoietic cell-or somatic stem cell-markers. Antigen-positive cells readily initiated neurosphere formation, but cells lacking these markers did so less readily. Doubly positive cells expressing both syndecan-1 and Notch-1 underwent neurosphere formation more efficiently than did singly positive cells. The progeny of sorted cells could differentiate into neurons and glial cells both in vitro and in vivo. These antibodies were also useful for isolating cells from the murine embryonic day 14.5 brain that efficiently formed neurospheres. In contrast, there was no distinct difference in neurosphere formation efficiency between Hoechst 33342-stained side population cells and main population cells, although the former are known to have a stem cell phenotype in various tissues. These results indicate the usefulness of syndecan-1, Notch-1, and integrin-β1 as NSC markers. © 2005 Wiley-Liss, Inc.

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Functional expression of ABCG2 transporter in human neural stem/progenitor cells

Islam Mohammed Omedul, Kanemura Yonehiro, Tajria Jesmin, Mori Hideki, Kobayashi Satoshi, Hara Masayuki, Yamasaki Mami, Okano Hideyuki, Miyake Jun

Neuroscience Research 52 （ 1 ） 75 - 82 2005年05月

ISSN 0168-0102

We have studied the expression, localization, and function of the ABCG2 transporter, a universal stem cell marker, at the protein level in human cultured neural stem/progenitor cells (hNSPCs) using immunoblotting, immunofluorescence, and ATPase assays. Human NSPCs were isolated from human fetal brain and propagated in vitro as neurospheres. Both the cells in neurospheres and single cells dissociated from neurospheres showed high levels of ABCG2, and about 63% of the cells in neurospheres were ABCG2-positive, similar to the proportion of nestin-positive cells, and in most cases the ABCG2 and nestin staining co-localized in the same cells. Both the three-dimensional structure of single hNSPCs stained with anti-ABCG2 antibodies and an examination using a biochemical marker for the plasma membrane indicated that ABCG2 was localized to the plasma membrane of hNSPCs. The ABCG2 expressed in hNSPCs had prazosin-sensitive ATP hydrolysis activity, and the ABCG2 level was sharply down-regulated during hNSPC differentiation. All these results suggested that ABCG2, was functionally expressed in hNSPCs. ABCG2 might play a significant role in maintaining human neural stem cells in an undifferentiated state and in protecting hNSPCs from xenobiotics or other toxic substances in vivo. © 2005 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

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Transplantation of human neural stem cells for spinal cord injury in primates

Iwanami A., Kaneko S., Nakamura M., Kanemura Y., Mori H., Kobayashi S., Yamasaki M., Momoshima S., Ishii H., Ando K., Tanioka Y., Tamaoki N., Nomura T., Toyama Y., Okano Hideyuki

Journal of Neuroscience Research 80 （ 2 ） 182 - 190 2005年04月

ISSN 0360-4012

Recent studies have shown that delayed transplantation of neural stem/progenitor cells (NSPCs) into the injured spinal cord can promote functional recovery in adult rats. Preclinical studies using nonhuman primates, however, are necessary before NSPCs can be used in clinical trials to treat human patients with spinal cord injury (SCI). Cervical contusion SCIs were induced in 10 adult common marmosets using a stereotaxic device. Nine days after injury, in vitro-expanded human NSPCs were transplanted into the spinal cord of five randomly selected animals, and the other sham-operated control animals received culture medium alone. Motor functions were evaluated through measurements of bar grip power and spontaneous motor activity, and temporal changes in the intramedullary signals were monitored by magnetic resonance imaging. Eight weeks after transplantation, all animals were sacrificed. Histologic analysis revealed that the grafted human NSPCs survived and differentiated into neurons, astrocytes, and oligodendrocytes, and that the cavities were smaller than those in sham-operated control animals. The bar grip power and the spontaneous motor activity of the transplanted animals were significantly higher than those of sham-operated control animals. These findings show that NSPC transplantation was effective for SCI in primates and suggest that human NSPC transplantation could be a feasible treatment for human SCI. © 2005 Wiley-Liss, Inc.

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Establishment of graded spinal cord injury model in a nonhuman primate

Iwanami A., Yamane J., Katoh H., Nakamura M., Momoshima S., Ishii H., Tanioka Y., Tamaoki N., Nomura T., Toyama Y., Okano Hideyuki

Journal of Neuroscience Research 80 （ 2 ） 172 - 181 2005年04月

ISSN 0360-4012

Most previous studies on spinal cord injury (SCI) have used rodent models. Direct extrapolation of the results obtained in rodents to clinical cases is difficult, however, because of neurofunctional and anatomic differences between rodents and primates. In the present study, the development of histopathologic changes and functional deficits were assessed quantitatively after mild, moderate, and severe spinal cord contusive injuries in common marmosets. Contusive SCI was induced by dropping one of three different weights (15, 17, or 20 g) at the C5 level from a height of 50 mm. Serial magnetic resonance images showed significant differences in the intramedullary T1 low signal and T2 high signal areas among the three groups. Quantitative histologic analyses revealed that the number of motor neurons, the myelinated areas, and the amounts of corticospinal tract fibers decreased significantly as the injury increased in severity. Motor functions were evaluated using the following tests: original behavioral scoring scale, measurements of spontaneous motor activity, bar grip test, and cage-climbing test. Significant differences in all test results were observed among the three groups. Spontaneous motor activities at 10 weeks after injury were closely correlated with the residual myelinated area at the lesion epicenter. The establishment of a reliable nonhuman primate model for SCI with objective functional evaluation methods should become an essential tool for future SCI treatment studies. Quantitative behavioral and histopathologic analyses enabled three distinct grades of injury severity (15-g, 17-g, and 20-g groups) to be characterized with heavier weights producing more serious injuries, and relatively constant behavioral and histopathologic outcomes. © 2005 Wiley-Liss, Inc.

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Functional properties of dopaminergic neurones in the mouse olfactory bulb

Pignatelli Angela, Kobayashi Kazuto, Okano Hideyuki, Belluzzi Ottorino

Journal of Physiology 564 （ 2 ） 501 - 514 2005年04月

ISSN 0022-3751

The olfactory bulb of mammals contains a large population of dopaminergic interneurones within the glomerular layer. Dopamine has been shown both in vivo and in vitro to modulate several aspects of olfactory information processing, but the functional properties of dopaminergic neurones have never been described due to the inability to recognize these cells in living preparations. To overcome this difficulty, we used a transgenic mouse strain harbouring an eGFP (enhanced green fluorescent protein) reporter construct under the promoter of tyrosine hydroxylase, the rate-limiting enzyme for cathecolamine synthesis. As a result, we were able to identify dopaminergic neurones (TH-GFP cells) in living preparations and, for the first time, we could study the functional properties of such neurones in the olfactory bulb, in both slices and dissociated cells. The most prominent feature of these cells was the autorhythmicity. In these cells we identified five main voltage-dependent conductances: the two having largest amplitude were a fast transient Na⁺ current and a delayed rectifier K⁺ current. In addition, we observed three smaller inward currents, sustained by Na⁺ ions (persistent type) and by Ca²⁺ ions (LVA and HVA). Using pharmacological tools and ion substitution methods we showed that the pacemaking process is supported by the interplay of the persistent Na⁺ current and of a T-type Ca²⁺ current. We carried out a complete kinetical analysis of the five conductances present in these cells, and developed a Hodgkin-Huxley model of TH-GFP cells, capable of reproducing accurately the properties of living cells, including autorhytmicity, and allowing a precise understanding of the process. © The Physiological Society 2005.

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Involvement of Hu and heterogeneous nuclear ribonucleoprotein K in neuronal differentiation through p21 mRNA post-transcriptional regulation

Yano Masato, Okano Hirotaka J., Okano Hideyuki

Journal of Biological Chemistry 280 （ 13 ） 12690 - 12699 2005年04月

ISSN 0021-9258

The Hu family is a group of neuronal RNA-binding proteins required for neuronal differentiation in the developing nervous system. Previously, Hu proteins have been shown to enhance the stabilization and/or translation of target mRNAs, such as p21 (CIP1), by binding to AU-rich elements in untranslated regions (UTRs). In this study, we show that Hu induces p21 expression, cell cycle arrest, and neuronal differentiation in mouse neuroblastoma N1E-115 cells. p21 expression is also up-regulated during Me₂SO-induced differentiation in N1E-115 cells and is controlled by post-transcriptional mechanisms through its 3′-UTR. To investigate the molecular mechanisms of Hu functions, we used a proteomics strategy to isolate Hu-interacting proteins and identified heterogeneous nuclear ribonucleoprotein (hnRNP) K. hnRNP K also specifically binds to CU-rich sequences in p21 mRNA 3′-UTR and represses its translation in both nonneuronal and neuronal cells. Further, using RNA interference experiments, we show that the Hu-p21 pathway contributes to the regulation of neurite outgrowth and proliferation in N1E-115 cells, and this pathway is antagonized by hnRNP K. Our results suggest a model in which the mutually antagonistic action of two RNA-binding proteins, Hu and hnRNP K, control the timing of the switch from proliferation to neuronal differentiation through the post-transcriptional regulation of p21 mRNA. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc.

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An oncolytic HSV-1 mutant expressing ICP34.5 under control of a nestin promoter increases survival of animals even when symptomatic from a brain tumor

Kambara Hirokazu, Okano Hideyuki, Chiocca E. Antonio, Saeki Yoshinaga

Cancer Research 65 （ 7 ） 2832 - 2839 2005年04月

ISSN 0008-5472

Oncolytic herpes simples virus-1 (HSV-1) mutants possessing mutations in the ICP34.5 and ICP6 genes have proven safe through clinical trials. However, ICP34.5-null viruses may grow poorly in cells due to their inability to prevent host-cell shut-off of protein synthesis caused by hyperphosphorylation of eukaryotic initiation factor 2α. To increase tumor selectivity, glioma-selective expression of ICP34.5 in the context of oncolysis may be useful. Malignant gliomas remain an incurable disease. One molecular marker of malignant gliomas is expression of the intermediate filament nestin. Expression of nestin mRNA was confirmed in 6 of 6 human glioma lines and in 3 of 4 primary glioma cells. Normal human astrocytes were negative. A novel glioma-selective HSV-1 mutant (rQNestin34.5) was thus engineered by expressing ICP34.5 under control of a synthetic nestin promoter. Replication, cellular propagation, and cytotoxicity of rQNestin34.5 were significantly enhanced in cultured and primary human glioma cell lines compared with control virus. However, replication, cellular propagation, and cytotoxicity of rQNestin34.5 in normal human astrocytes remained quantitatively similar to that of control virus. In glioma cell lines infected with rQNestin34.5, the level of phospho-eukaryotic initiation factor 2α was lower than that of cells infected by control rHsvQ1, confirming selective ICP34.5 expression in glioma cells. In vivo, rQNestin34.5 showed significantly more potent inhibition of tumor growth compared with control virus. Treatment in the brain tumor model was instituted on animal's display of neurologic symptoms, which usually led to rapid demise. rQNestin34.5 treatment doubled the life span of these animals. These results show that rQNestin34.5 could be a potent agent for the treatment of malignant glioma. ©2005 American Association for Cancer Research.

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A novel marker for Purkinje cells, KIAA0864 protein. An analysis based on a monoclonal antibody HFB-16 in developing human cerebellum

Nakamura Yasuhiro, Yamamoto Munehiko, Oda Eriko, Kanemura Yonehiro, Kodama Eri, Yamamoto Atsuyo, Yamamoto Hideyuki, Miyado Kenji, Okano Hirotaka James, Fukagawa Ryouji, Higaki Koichi, Yamasaki Mami, Okano Hideyuki

Journal of Histochemistry and Cytochemistry 53 （ 4 ） 423 - 430 2005年04月

ISSN 0022-1554

In the search for immunohistochemical markers of the developing human brain, a monoclonal antibody, HFB-16, was raised against homogenates from the cerebrum of a 15-gestational-week-old (GW) human fetus and screened on paraffin-embedded human embryonic brain specimens. This antibody was particularly useful as a marker for Purkinje cells in the developing human cerebellum. Positive immunoreactivities with HFB-16 first appeared in the Purkinje cell layer at 17 GW. From 20 to 24 GW, positive immunoreactivities were found above the lamina dissecans. After 25 GW, dendrites of Purkinje cells were found with the HFB-16 antibody, and the nerve fibers of the Purkinje cells became positive after 35 GW. Neurons in the dentate nucleus and external and internal granular layers reacted negatively to this antibody. After 1 year, when the external granular layer faded out, the dendrites of the Purkinje cells reached the pial surface of the cerebellum, and nerve fibers began to develop in the white matter. This antibody was also useful for characterization of components in heterotopic neurons found in various anomaly syndromes such as trisomy 13. Expressional cloning indicated the antigen against HFB-16 to be humain KIAA0864 protein, which is supposed to be an alternative splicing product of p116Rip, whose function has not yet been elucidated. The antigenicity of the KIAA0864 protein was confirmed using human cDNA of the KIAA0864 protein, a protein expression vector, and an HFB-16 antibody.

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Components of the transcriptional mediator complex are required for asymmetric cell division in C. elegans

Yoda Akinori, Kouike Hiroko, Okano Hideyuki, Sawa Hitoshi

Development 132 （ 8 ） 1885 - 1893 2005年04月

ISSN 0950-1991

Asymmetric cell division is a fundamental process that produces cellular diversity during development. In C. elegans, the Wnt signaling pathway regulates the asymmetric divisions of a number of cells including the T blast cell. We found that the let-19 and dpy-22 mutants have defects in their T-cell lineage, and lineage analyses showed that the defects were caused by disruption in the asymmetry of the T-cell division. We found that let-19 and dpy-22 encode homologs of the human proteins MEDD13/TRAP240 and MED12/TRAP230, respectively, which are components of the Mediator complex. Mediator is a multi-component complex that can regulate transcription by transducing the signals between activators and RNA polymerase in vitro. We also showed that LET-19 and DPY-22 form a complex in vivo with other components of Mediator, SUR-2/MED23 and LET-425/MED6. In the let-19 and dpy-22 mutants, tlp-1, which is normally expressed asymmetrically between the T-cell daughters through the function of the Wnt pathway, was expressed symmetrically in both daughter cells. Furthermore, we found that the let-19 and dpy-22 mutants were defective in the fusion of the Pn.p cell, a process that is regulated by bar-1/β-catenin. Ectopic cell fusion in bar-1 mutants was suppressed by the let-19 or dpy-22 mutations, while defective cell fusion in let-19 mutants was suppressed by lin-39/Hox mutations, suggesting that let-19 and dpy-22 repress the transcription of lin-39. These results suggest that LET-19 and DPY-22 in the Mediator complex repress the transcription of Wnt target genes.

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Lecithinized brain-derived neurotrophic factor promotes the differentiation of embryonic stem cells in vitro and in vivo

Kitagawa Aki, Nakayama Toshiaki, Takenaga Mitsuko, Matsumoto Kayo, Tokura Yukie, Ohta Yuki, Ichinohe Manabu, Yamaguchi Yoko, Suzuki Noboru, Okano Hideyuki, Igarashi Rie

Biochemical and Biophysical Research Communications 328 （ 4 ） 1051 - 1057 2005年03月

ISSN 0006-291X

The addition of lecithin molecules to brain-derived neurotrophic factor (BDNF) has been reported to markedly enhance its pharmacological effect in vivo. In the current study, we show that lecithinized BDNF (PC-BDNF) has a higher affinity than BDNF for neural precursor cells. Although BDNF only slightly increased the expression of the genes for Mash-1, p35, 68 kDa neurofilament, and TrkB receptor, PC-BDNF caused a significant increase in their expression. PC-BDNF also increased the level of neurofilament protein and dramatically increased TrkB mRNA gene expression, which was followed by a sustained activation of the p42/p44 extracellular-regulated kinases. Finally, transplantation of PC-BDNF-treated cells was more effective than BDNF-treated cells at improving impaired motor function caused by spinal cord injury. These findings showed that PC-BDNF has a better potential than BDNF for promoting neural differentiation, partly due to a higher cellular affinity. Furthermore, PC-BDNF-treated cells could be useful for transplantation therapy for central nervous system injuries. © 2005 Elsevier Inc. All rights reserved.

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Loss of imprinting of Igf2 alters intestinal maturation and tumorigenesis in mice

Sakatani Takashi, Kaneda Atsushi, Iacobuzio-Donahue Christine A., Carter Mark G., De Boom Witzel Sten, Okano Hideyuki, Ko Minora S H, Ohlsson Rolf, Longo Dan L., Feinberg Andrew P.

Science 307 （ 5717 ） 1976 - 1978 2005年03月

ISSN 0036-8075

Loss of imprinting (LOI) of the insulin-like growth factor II gene (IGF2) is an epigenetic alteration that results in a modest increase in IGF2 expression, and it is present in the normal colonic mucosa of about 30% of patients with colorectal cancer. To investigate its role in intestinal tumorigenesis, we created a mouse model of Igf2 LOI by crossing female H19 ^+/- mice with male Apc^+/Min mice. Mice with LOI developed twice as many intestinal tumors as did control littermates. Notably, these mice also showed a shift toward a less differentiated normal intestinal epithelium, reflected by an increase in crypt length and increased staining with progenitor cell markers. A similar shift in differentiation was seen in the normal colonic mucosa of humans with LOI. Thus, altered maturation of nonneoplastic tissue may be one mechanism by which epigenetic changes affect cancer risk.

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The RNA-binding protein HuD regulates neuronal cell identity and maturation

Akamatsu Wado, Fujihara Hiroaki, Mitsuhashi Takayuki, Yano Masato, Shibata Shinsuke, Hayakawa Yoshika, Okano Hirotaka James, Sakakibara Shin Ichi, Takano Hiroshi, Takano Toshiya, Takahashi Takao, Noda Tetsuo, Okano Hideyuki

Proceedings of the National Academy of Sciences of the United States of America 102 （ 12 ） 4625 - 4630 2005年03月

ISSN 0027-8424

Neural Hu proteins (HuB/C/D) are RNA-binding proteins that have been shown to induce neuronal differentiation activity when overexpressed in immature neural progenitor cells or undifferentiated neuronal tumors. Newly generated HuD-deficient mice exhibited a transient impaired-cranial-nerve-development phenotype at an early embryonic stage. Adult HuD-deficient mice exhibited an abnormal hind-limb reflex and poor rotarod performance. Analysis of neurosphere formation revealed that the number and self-renewal capacity of the neural stem/progenitor cells were increased in HuD-deficient mice. HuD-deficient primary neurospheres also generated a smaller number of neurons. Cohort analysis of the cellular proliferative activity by using BrdUrd and iododeoxuridine labeling revealed that the number of differentiating quiescent cells in the embryonic cerebral wall was decreased. Long-term administration of BrdUrd revealed that the number of slowly dividing stem cells in the adult subventricular zone was increased in the HuD-deficient mice. Taken together, the results suggest that HuD is required at multiple points during neuronal development, including negative regulation of proliferative activity and neuronal cell-fate acquisition of neural stem/progenitor cells. © 2005 by The National Academy of Sciences of the USA.

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Hes1-deficient mice show precocious differentiation of Paneth cells in the small intestine

Suzuki Katsumasa, Fukui Hirokazu, Kayahara Takahisa, Sawada Mitsutaka, Seno Hiroshi, Hiai Hiroshi, Kageyama Ryoichiro, Okano Hideyuki, Chiba Tsutomu

Biochemical and Biophysical Research Communications 328 （ 1 ） 348 - 352 2005年03月

ISSN 0006-291X

We have previously shown that Hes1 is expressed both in putative epithelial stem cells just above Paneth cells and in the crypt base columnar cells between Paneth cells, while Hes1 is completely absent in Paneth cells. This study was undertaken to clarify the role of Hes1 in Paneth cell differentiation, using Hes1-knockout (KO) newborn (P0) mice. Electron microscopy revealed premature appearance of distinct cells containing cytoplasmic granules in the intervillous region in Hes1-KO P0 mice, whereas those cells were absent in wild-type (WT) P0 mice. In Hes1-KO P0 mice, the gene expressions of cryptdins, exclusively present in Paneth cells, were all enhanced compared with WT P0 mice. Immunohistochemistry demonstrated increased number of both lysozyme-positive and cryptdin-4-positive cells in the small intestinal epithelium of Hes1-KO P0 mice as compared to WT P0 mice. Thus, Hes1 appears to have an inhibitory role in Paneth cell differentiation in the small intestine. © 2005 Elsevier Inc. All rights reserved.

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Leukemia inhibitory factor induces multi-lineage differentiation of adult stem-like cells in kidney via kidney-specific cadherin 16

Hishikawa Keiichi, Marumo Takeshi, Miura Shigeki, Nakanishi Asato, Matsuzaki Yumi, Shibata Katsunori, Kohike Hiroko, Komori Takuya, Hayashi Matsuhiko, Nakaki Toshio, Nakauchi Hiromitsu, Okano Hideyuki, Fujita Toshiro

Biochemical and Biophysical Research Communications 328 （ 1 ） 288 - 291 2005年03月

ISSN 0006-291X

Side population (SP) is reported to be a stem cell-rich population. In the presence of leukemia inhibitory factor (LIF), cultured kidney SP cells differentiated into multi-lineage in collagen gel but not in synthesized polymer that has no cell adhesion factor. In cultured kidney SP cells, gene expression of kidney-specific cadherin 16 was specifically upregulated in collagen gel but not in synthesized polymer. Moreover, decreasing cadherin 16 expression using siRNA abolished LIF-induced multi-lineage differentiation of kidney SP in collagen gel. These results indicated that LIF induced multi-lineage differentiation of adult stem-like cells in kidney via cadherin 16. © 2005 Elsevier Inc. All rights reserved.

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Erratum

Sakuragawa Norio, Kakinuma Kenichi, Kikuchi Aiko, Okano Hideyuki, Uchida Saiko, Kamo Isao, Kobayashi Mamoru, Yokoyama Yasunobu

Journal of Neuroscience Research 79 （ 5 ） 725 2005年03月

ISSN 0360-4012

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Epigenetics, I GF2 dosage and cancer risk

Kaneda [No Value], Feinberg [No Value], Iacobuzio-Donahue [No Value], Sakatani Takashi, Witzel Sten De Boom, Carter Mark, Ko Minoru, Longo Dan, Okano Hideyuki, Ohlsson [No Value]

Cancer Biology and Therapy 4 （ 3 ） 2005年03月

ISSN 1538-4047

Comparative functional genomics revealed conservation and diversification of three enhancers of the isl1 gene for motor and sensory neuron-specific expression

Uemura Osamu, Okada Yohei, Ando Hideki, Guedj Mickael, Higashijima Shin Ichi, Shimazaki Takuya, Chino Naoichi, Okano Hideyuki, Okamoto Hitoshi

Developmental Biology 278 （ 2 ） 587 - 606 2005年02月

ISSN 0012-1606

Islet-1 (Isl1) is a member of the Isl1 family of LIM-homeodomain transcription factors (LIM-HD) that is expressed in a defined subset of motor and sensory neurons during vertebrate embryogenesis. To investigate how this specific expression of isl1 is regulated, we searched for enhancers of the isl1 gene that are conserved in vertebrate evolution. Initially, two enhancer elements, CREST1 and CREST2, were identified downstream of the isl1 locus in the genomes of fugu, chick, mouse, and human by BLAST searching for highly similar elements to those originally identified as motor and sensory neuron-specific enhancers in the zebrafish genome. The combined action of these elements is sufficient for completely recapitulating the subtype-specific expression of the isl1 gene in motor neurons of the mouse spinal cord. Furthermore, by direct comparison of the upstream flanking regions of the zebrafish and human isl1 genes, we identified another highly conserved noncoding element, CREST3, and subsequently C3R, a similar element to CREST3 with two CDP CR1 recognition motifs, in the upstream regions of all other isl1 family members. In mouse and human, CRESTs are located as far as more than 300 kb away from the isl1 locus, while they are much closer to the isl1 locus in zebrafish. Although all of zebrafish CREST2, CREST3, and C3R activate gene expression in the sensory neurons of zebrafish, CREST2 of mouse and human does not have the sequence necessary for sensory neuron-specific expression. Our results revealed both a remarkable conservation of the regulatory elements regulating subtype-specific gene expression in motor and sensory neurons and the dynamic process of reorganization of these elements whereby each element increases the level of cell-type specificity by losing redundant functions with the other elements during vertebrate evolution. © 2004 Elsevier Inc. All rights reserved.

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Regenerative medicine for the central nervous system - Transplantation of neural stem cells into the injured spinal cord -

Kaneko Shinjiro, Nakamura Masaya, Ogawa Yuto, Watanabe Kota, Iwanami Akio, Toyama Yoshiaki, Okano Hideyuki

Japan Medical Association Journal 48 （ 2 ） 68 - 80 2005年02月

ISSN 1346-8650

Advances in the stem cell biology of the central nervous system (CNS) and other studies enhance the possibility of regeneration of the damaged CNS, which has previously been considered to be almost impossible. The availability of signals to induce the appropriate differentiation of the transplanted and/or endogenous neural stem cells (NSCs) as well as the timing of the transplantation are vital factors for improving the functional recovery of the damaged CNS. Because the immediate post-traumatic microenvironment of the spinal cord is in an acute inflammatory stage, it does not favor the survival and differentiation of the transplants. Furthermore, in the chronic disease stage after injury, glial scars that inhibit the regeneration of neuronal axons form in the injured site. Thus, we consider that the optimal period of transplantation is 1-2 weeks after injury.

The transmembrane protein, Tincar, is involved in the development of the compound eye in Drosophila melanogaster

Hirota Yuki, Sawamoto Kazunobu, Takahashi Kuniaki, Ueda Ryu, Okano Hideyuki

Development Genes and Evolution 215 （ 2 ） 90 - 96 2005年02月

ISSN 0949-944X

We previously cloned and characterized the Drosophila gene, tincar (tinc), which encodes a novel protein with eight putative transmembrane domains. Here, we have studied the expression pattern and functions of tinc during developmental processes. Tinc mRNA is expressed in the central and peripheral nervous systems, and midgut during embryogenesis. In the third-instar larval eye disc, tinc mRNA is strongly expressed in all the differentiating ommatidial cells within and in the vicinity of the morphogenetic furrow. Loss-of-function analysis using the RNA-interference method revealed severe defects of eye morphogenesis during the late developmental stages. Our results suggested that tinc may have an indispensable role in the normal differentiation of ommatidial cells. © Springer-Verlag 2005.

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A putative human breast stem cell population is enriched for steroid receptor-positive cells

Clarke Robert B., Spence Katherine, Anderson Elizabeth, Howell Anthony, Okano Hideyuki, Potten Christopher S.

Developmental Biology 277 （ 2 ） 443 - 456 2005年01月

ISSN 0012-1606

Breast epithelial stem cells are thought to be the primary targets in the etiology of breast cancer. Since breast cancers mostly express estrogen and progesterone receptor (ERα and PR), we examined the biology of these ERα/PR-positive cells and their relationship to stem cells in normal human breast epithelium. We employed several complementary approaches to identify putative stem cell markers, to characterise an isolated stem cell population and to relate these to cells expressing the steroid receptors ERα and PR. Using DNA radiolabelling in human tissue implanted into athymic nude mice, a population of label-retaining cells were shown to be enriched for the putative stem cell markers p21 ^CIP1 and Msi-1, the human homolog of Drosophila Musashi. Steroid receptor-positive cells were found to co-express these stem cell markers together with cytokeratin 19, another putative stem cell marker in the breast. Human breast epithelial cells with Hoechst dye-effluxing "side population" (SP) properties characteristic of mammary stem cells in mice were demonstrated to be undifferentiated "intermediate" cells by lack of expression of myoepithelial and luminal apical membrane markers. These SP cells were 6-fold enriched for ERα-positive cells and expressed several fold higher levels of the ERα, p21 ^CIP1 and Msi1 genes than non-SP cells. In contrast to non-SP cells, SP cells formed branching structures in matrigel which included cells of both luminal and myoepithelial lineages. The data suggest a model where scattered steroid receptor-positive cells are stem cells that self-renew through asymmetric cell division and generate patches of transit amplifying and differentiated cells. © 2004 Elsevier Inc. All rights reserved.

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Transient inhibition of BMP signaling by Noggin induces cardiomyocyte differentiation of mouse embryonic stem cells

Yuasa Shinsuke, Itabashi Yuji, Koshimizu Uichi, Tanaka Tomofumi, Sugimura Keijiro, Kinoshita Masayoshi, Hattori Fumiyuki, Fukami Shin Ichi, Shimazaki Takuya, Okano Hideyuki, Ogawa Satoshi, Fukuda Keiichi

Nature Biotechnology 23 （ 5 ） 607 - 611 2005年

ISSN 1087-0156

Embryonic stem (ES) cells are a promising source of cardiomyocytes, but clinical application of ES cells has been hindered by the lack of reliable selective differentiation methods. Differentiation into any lineage is partly dependent on the regulatory mechanisms of normal early development. Although several signals, including bone morphogenetic protein (BMP), Wnt and FGF, are involved in heart development, scarce evidence is available about the exact signals that mediate cardiomyocyte differentiation. While investigating the involvement of BMP signaling in early heart formation in the mouse, we found that the BMP antagonist Noggin is transiently but strongly expressed in the heart-forming region during gastrulation and acts at the level of induction of mesendoderm to establish conditions conducive to cardiogenesis. We applied this finding to develop an effective protocol for obtaining cardiomyocytes from mouse ES cells by inhibition of BMP signaling.

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Erratum

Yuasa Shinsuke, Itabashi Yuji, Koshimizu Uichi, Tanaka Tomofumi, Sugimura Keijiro, Kinoshita Masayoshi, Hattori Fumiyuki, Fukami Shin Ichi, Shimazaki Takuya, Ogawa Satoshi, Okano Hideyuki, Fukuda Keiichi

Nature Biotechnology 23 （ 7 ） 897 2005年

ISSN 1087-0156

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In vitro screening of exogenous factors for human neural stem/progenitor cell proliferation using measurement of total ATP content in viable cells

Kanemura Yonehiro, Mori Hideki, Nakagawa Atsuyo, Islam Mohammed Omedul, Kodama Eri, Yamamoto Atsuyo, Shofuda Tomoko, Kobayashi Satoshi, Miyake Jun, Yamazaki Tomohiko, Hirano Shun Ichiro, Yamasaki Mami, Okano Hideyuki

Cell Transplantation 14 （ 9 ） 673 - 682 2005年

ISSN 0963-6897

One of the newest and most promising methods for treating intractable neuronal diseases and injures is the transplantation of ex vivo-expanded human neural stem/progenitor cells (NSPCs). Human NSPCs are selectively expanded as free-floating neurospheres in serum-free culture medium containing fibroblast growth factor 2 (FGF2) and/or epidermal growth factor (EGF); however, the culture conditions still need to be optimized for performance and cost before the method is used clinically. Here, to improve the NSPC culture method for clinical use, we used an ATP assay to screen the effects of various reagents on human NSPC proliferation. Human NSPCs responded to EGF, FGF2, and leukemia inhibitory factor (LIF) in a dose-dependent manner, and the minimum concentrations eliciting maximum effects were 10 ng/ml EGF, 10 ng/ ml FGF2, and 5 ng/ml LIF. EGF and LIF were stable in culture medium without NSPCs, although FGF2 was degraded. In the presence of human NSPCs, however, FGF2 and LIF were both degraded very rapidly, to below the estimated minimum concentration on day 3, but EGF remained above the minimum concentration for 5 days. Adding supplemental doses of each growth factor during the incubation promoted human NSPC proliferation. Among other supplements, insulin and transferrin promoted human NSPC growth, but progesterone, putrescine, selenite, D-glucose, and lactate were not effective and were cytotoxic at higher concentrations. Supplementing with conditioned medium from human NSPCs significantly increased human NSPC proliferation, but using a high percentage of the medium had a negative effect. These findings suggest that human NSPC culture is regulated by a balance in the culture medium between decreasing growth factor levels and increasing positive or negative factors derived from the NSPCs. Thus, in designing culture conditions for human NSPCs, it is useful to take the individual properties of each factor into consideration. Copyright © 2005 Cognizant Comm. Corp.

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In vivo PET measurements with [¹¹C]PE2I to evaluate fetal mesencephalic transplantations to unilateral 6-OHDA-lesioned rats

Inaji Motoki, Yoshizaki Takahito, Okauchi Takashi, Maeda Jun, Nagai Yuji, Nariai Tadashi, Ohno Kikuo, Ando Kiyoshi, Okano Hideyuki, Obayashi Shigeru, Suhara Tetsuya

Cell Transplantation 14 （ 9 ） 655 - 663 2005年

ISSN 0963-6897

Positron emission tomography (PET) is a useful tool to assess and visualize neurotransmissions in vivo. In this study, we performed repeated PET scans with [¹¹C]PE2I, a tracer of the dopamine transporter, to evaluate the alteration of the expression of dopamine (DA) transmission component after a fetal mesencephalic transplantation. The fetal mesencephalic cells were transplanted into the striatum of unilateral 6-OHDA-lesioned rats. PET scans with [¹¹C]PE2I were performed to evaluate the DA transporter before and 2 and 4 weeks after the transplantation. Rotation behavior tests, in vitro autoradiography, measurements of DA contents in the striatum by high-performance liquid chromatography (HPLC), and tyrosine hydroxylase (TH) immunohistological examinations were performed at the same time points and examined for their relationship to changes in the dopamine transporter. The number of ipsilateral rotations induced by methamphetamine injections decreased. DA contents in the striatum measured with HPLC significantly increased. In the PET study, the binding potential of [¹¹C]PE2I increased at 4 weeks. The results of the in vitro autoradiography study corresponded with those of the PET study. The degrees of the change in the binding potentials correlated with those of the numbers of rotations in the behavioral study and the DA contents in the striatum. In the histological examination, TH-positive cells with axons were observed at 2 and 4 weeks after the transplantation. As the dopamine transporter exists only in the axon terminal of DA neurons, these results suggested that PET measurements of [¹¹C]PE2I binding indicated not only survival, but maturity and functioning of the transplanted cells. Repeated PET measurements of DA transporters are a useful tool in assessing the effectiveness of neural transplantations. Copyright © 2005 Cognizant Comm. Corp.

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Effect of neurosphere size on the growth rate of human neural stem/progenitor cells

Mori Hideki, Ninomiya Kazuaki, Kino-Oka Masahiro, Shofuda Tomoko, Islam Mohammed Omedul, Yamasaki Mami, Okano Hideyuki, Taya Masahito, Kanemura Yonehiro

Journal of Neuroscience Research 84 （ 8 ） 1682 - 1691 2006年12月

ISSN 0360-4012

Neural stem/progenitor cells (NSPCs) proliferate as aggregates in vitro, but the mechanism of aggregation is not fully understood. Here, we report that aggregation promotes the proliferation of NSPCs. We found that the proliferation rate was linear and depended on the size of the aggregate; that is, the population doubling time of the NSPCs gradually decreased as the diameter approached 250 μm and flattened to a nearly constant value beyond this diameter. Given this finding, and with the intent of enhancing the efficiency of human NSPC expansion, we induced the NSPCs to form aggregates close to 250 μm in diameter quickly by culturing them in plates with U-bottomed wells. The NSPCs formed aggregates effectively in the U-bottomed wells, with cell numbers approximately 1.5 times greater than those in the aggregates that formed spontaneously in flat-bottomed wells. In addition, this effect of aggregation involved cell-cell signaling molecules of the Notch1 pathway. In the U-bottomed wells, Hes1 and Hes5, which are target genes of the Notch signal, were expressed at higher levels than in the control, flat-bottomed wells. The amount of cleaved Notch1 was also higher in the cells cultured in the U-bottomed wells. The addition of γ-secretase inhibitor, which blocks Notch signaling, suppressed cell proliferation in the U-bottomed wells. These results suggest that the three-dimensional architecture of NSPC aggregates would create a microenvironment that promotes the proliferation of human NSPCs. © 2006 Wiley-Liss, Inc.

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Neuroprotective effects of angiotensin II type 1 receptor (AT1R) blocker, telmisartan, via modulating AT1R and AT2R signaling in retinal inflammation

岡野栄之

INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE 47 （ 12 ） 5545 - 5552 2006年12月

研究論文（学術雑誌）, ISSN 0146-0404

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Neutralization of ciliary neurotrophic factor reduces astrocyte production from transplanted neural stem cells and promotes regeneration of corticospinal tract fibers in spinal cord injury

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 84 （ 8 ） 1669 - 1681 2006年12月

研究論文（学術雑誌）, ISSN 0360-4012

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An oncolytic HSV1 mutant expressing ICP34.5 under control of a nestin promoter increases survival of animals, even when symptomatic from a brain tumor

岡野栄之

JOURNAL OF GENE MEDICINE 8 （ 12 ） 1453 - 1453 2006年12月

研究論文（学術雑誌）, ISSN 1099-498X

Expression of Musashi-1 in normal and regenerating retinas of adult newt

岡野栄之

ZOOLOGICAL SCIENCE 23 （ 12 ） 1180 - 1180 2006年12月

研究論文（学術雑誌）, ISSN 0289-0003

A practical device for pinpoint delivery of molecules into multiple neurons in culture

岡野栄之

BRAIN CELL BIOLOGY 35 （ 4-6 ） 229 - 237 2006年12月

研究論文（学術雑誌）, ISSN 1559-7105

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Effects of a moderate-intensity static magnetic field on VEGF-A stimulated endothelial capillary tubule formation in vitro

岡野栄之

BIOELECTROMAGNETICS 27 （ 8 ） 628 - 640 2006年12月

研究論文（学術雑誌）, ISSN 0197-8462

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Isolation of multipotent neural crest-derived stem cells from the adult mouse cornea

岡野栄之

STEM CELLS 24 （ 12 ） 2714 - 2722 2006年12月

研究論文（学術雑誌）, ISSN 1066-5099

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A selective Sema3A inhibitor enhances regenerative responses and functional recovery of the injured spinal cord

岡野栄之

NATURE MEDICINE 12 （ 12 ） 1380 - 1389 2006年12月

研究論文（学術雑誌）, ISSN 1078-8956

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Roles of RNA binding proteins in neural development

Yano Masato, Okano Hirotaka James, Okano Hideyuki

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 51 （ 16 Suppl ） 2603 - 2608 2006年12月

ISSN 0039-9450

Bidirectional regulation of dopamine D2 and neurotensin NTS1 receptors in dopamine neurons

Jomphe C., Lemelin P. L., Okano H., Kobayashi K., Trudeau L. E.

European Journal of Neuroscience 24 （ 10 ） 2789 - 2800 2006年12月

ISSN 0953-816X

Several lines of evidence suggest a close association between dopamine (DA) and neurotensin (NT) systems in the CNS. Indeed, in the rodent brain, abundant NT-containing fibres are found in DA-rich areas such as the ventral tegmental area and substantia nigra. Moreover, it has been shown in vivo that NT, acting through its high-affinity receptor (NTS1), reduces the physiological and behavioural effects of DA D2 receptor (D2R) activation, a critical autoreceptor feedback system regulating DA neurotransmission. However, the mechanism of this interaction is still elusive. The aim of our study was thus to reproduce in vitro the interaction between D2R and NTS1, and then to characterize the mechanisms implicated. We used a primary culture model of DA neurons prepared from transgenic mice expressing green fluorescent protein under the control of the tyrosine hydroxylase promoter. In these cultures, DA neurons endogenously express both D2R and NTS1. Using electrophysiological recordings, we show that activation of D2R directly inhibits the firing rate of DA neurons. In addition, we find that NT, acting through a NTS1-like receptor, is able to reduce D2R autoreceptor function independently of its ability to enhance DA neuron firing, and that this interaction occurs through a protein kinase C- and Ca ²⁺-dependent mechanism. Furthermore, prior activation of D2R reduces the ability of NTS1 to induce intracellular Ca²⁺ mobilization. Our findings provide evidence for bidirectional interaction between D2R and NTS1 in DA neurons, a regulatory mechanism that could play a key role in the control of the activity of these neurons. © The Authors (2006).

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Generation of neural cells from embryonic stem cells

Shimazaki Takuya, Okada Yohei, Yoshizaki Takahito, Okano Hideyuki

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 51 （ 13 ） 1854 - 1861 2006年11月

ISSN 0039-9450

Enhanced therapeutic efficacy of oncotytic herpes vector G207 against human non-small cell lung cancer - expression of an RNA-binding protein, Musashi1, as a marker for the tailored gene therapy

岡野栄之

JOURNAL OF GENE MEDICINE 8 （ 11 ） 1329 - 1340 2006年11月

研究論文（学術雑誌）, ISSN 1099-498X

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Bcl2 enhances survival of newborn neurons in the normal and ischemic hippocampus

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 84 （ 6 ） 1187 - 1196 2006年11月

研究論文（学術雑誌）, ISSN 0360-4012

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Musashi1 antigen expression in human fetal germinal matrix development

岡野栄之

EXPERIMENTAL NEUROLOGY 201 （ 2 ） 515 - 518 2006年10月

研究論文（学術雑誌）, ISSN 0014-4886

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Role of the cholinergic system in regulating survival of newborn neurons in the adult mouse dentate gyrus and olfactory bulb

岡野栄之

GENES TO CELLS 11 （ 10 ） 1145 - 1159 2006年10月

研究論文（学術雑誌）, ISSN 1356-9597

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Integrity of plasmid deoxyribonucleic acid after application of laser-induced stress waves used for gene transfection

岡野栄之

JAPANESE JOURNAL OF APPLIED PHYSICS PART 2-LETTERS & EXPRESS LETTERS 45 （ 29-32 ） L768 - L769 2006年08月

研究論文（学術雑誌）, ISSN 0021-4922

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Ghrelin and leptin reciprocally regulate NPY neurons in the arcuate nucleus

岡野栄之

REGULATORY PEPTIDES 135 （ 3 ） 136 - 137 2006年08月

研究論文（学術雑誌）, ISSN 0167-0115

Mapping of Notch activation during cochlear development in mice

Murata Junko, Tokunaga Akinori, Okano Hideyuki, Kubo Takeshi

Journal of Comparative Neurology 497 （ 3 ） 502 - 518 2006年07月

ISSN 0021-9967

Recent chick experiments have shown that Notch signaling plays context-dependent distinct roles in inner ear development: initially, Notch activity confers a prosensory character on groups of cells by "lateral induction"; subsequently, it is involved in the establishment of fine-graded patterns of hair cells and supporting cells by "lateral inhibition." However, the spatiotemporal pattern of Notch activation in situ during mammalian inner ear development has not been investigated. In this study, we detected the expression patterns of the activated form of Notch1 (actN1) as well as those of endogenous Notch1, Jagged1 (Jag1), and Math1. ActN1 was detected by immunohistochemistry using an antibody that specifically recognizes the processed form of the intracellular domain of Notch1 cleaved by presenilin/γ-secretase activity. Between embryonic days (E)12.5 and E14.5, actN1 was weakly detected mainly in the medial region of cochlear epithelium, where Jag1-immunoreactivivty (IR) was also observed. Jag1-IR gradually became stronger in a more sharply defined area, finally becoming localized in supporting cells, while actN1 was detected in an overlapping area. Thus, a positive feedback loop was assumed to exist between the expression of Jag1 and actN1. In addition, actN1 started to be strongly expressed in the cells surrounding Math1-positive hair cell progenitors between E14.5 and E15.5. Strong actN1-IR continued in both a supporting cell lineage and in the greater epithelial ridge during the perinatal stage but ended by P7, suggesting that Notch1 activation may initially demarcate a prosensory region in the cochlear epithelium and then inhibit progenitor cells from becoming hair cells via classical "lateral inhibition." © 2006 Wiley-Liss, Inc.

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Wnt Signaling and a Hox Protein Cooperatively Regulate PSA-3/Meis to Determine Daughter Cell Fate after Asymmetric Cell Division in C. elegans

Arata Yukinobu, Kouike Hiroko, Zhang Yanping, Herman Michael A., Okano Hideyuki, Sawa Hitoshi

Developmental Cell 11 （ 1 ） 105 - 115 2006年07月

ISSN 1534-5807

Asymmetric cell division is a mechanism for achieving cellular diversity. In C. elegans, many asymmetric cell divisions are controlled by the Wnt-MAPK pathway through POP-1/TCF. It is poorly understood, however, how POP-1 determines the specific fates of daughter cells. We found that nob-1/Hox, ceh-20/Pbx, and a Meis-related gene, psa-3, are required for asymmetric division of the T hypodermal cell. psa-3 expression was asymmetric between the T cell daughters, and it was regulated by POP-1 through a POP-1 binding site in the psa-3 gene. psa-3 expression was also regulated by NOB-1 and CEH-20 through a NOB-1 binding sequence in a psa-3 intron. PSA-3 can bind CEH-20 and function after the T cell division to promote the proper fate of the daughter cell. These results indicate that cooperation between Wnt signaling and a Hox protein functions to determine the specific fate of a daughter cell. © 2006 Elsevier Inc. All rights reserved.

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Wnt signaling and a Hox protein cooperatively regulate PSA-3/Meis to determine daughter cell fate after asymmetric cell division in C-elegans

岡野栄之

DEVELOPMENTAL CELL 11 （ 1 ） 105 - 115 2006年07月

研究論文（学術雑誌）, ISSN 1534-5807

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Enhanced neurogenesis in the ischemic striatum following EGF-induced expansion of transit-amplifying cells in the subventricular zone

岡野栄之

NEUROSCIENCE LETTERS 403 （ 1-2 ） 63 - 67 2006年07月

研究論文（学術雑誌）, ISSN 0304-3940

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Conditional ablation of Stat3 or Socs3 discloses a dual role for reactive astrocytes after spinal cord injury

岡野栄之

NATURE MEDICINE 12 （ 7 ） 829 - 834 2006年07月

研究論文（学術雑誌）, ISSN 1078-8956

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Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice

Hara Taichi, Nakamura Kenji, Matsui Makoto, Yamamoto Akitsugu, Nakahara Yohko, Suzuki-Migishima Rika, Yokoyama Minesuke, Mishima Kenji, Saito Ichiro, Okano Hideyuki, Mizushima Noboru

Nature 441 （ 7095 ） 885 - 889 2006年06月

ISSN 0028-0836

Autophagy is an intracellular bulk degradation process through which a portion of the cytoplasm is delivered to lysosomes to be degraded. Although the primary role of autophagy in many organisms is in adaptation to starvation, autophagy is also thought to be important for normal turnover of cytoplasmic contents, particularly in quiescent cells such as neurons. Autophagy may have a protective role against the development of a number of neurodegenerative diseases. Here we report that loss of autophagy causes neurodegeneration even in the absence of any disease-associated mutant proteins. Mice deficient for Atg5 (autophagy-related 5) specifically in neural cells develop progressive deficits in motor function that are accompanied by the accumulation of cytoplasmic inclusion bodies in neurons. In Atg5^-/- cells, diffuse, abnormal intracellular proteins accumulate, and then form aggregates and inclusions. These results suggest that the continuous clearance of diffuse cytosolic proteins through basal autophagy is important for preventing the accumulation of abnormal proteins, which can disrupt neural function and ultimately lead to neurodegeneration. © 2006 Nature Publishing Group.

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Dynamic temporal and cell type-specific expression of Wnt signaling components in the developing midbrain

Rawal Nina, Castelo-Branco Gonçalo, Sousa Kyle M., Kele Julianna, Kobayashi Kazuto, Okano Hideyuki, Arenas Ernest

Experimental Cell Research 312 （ 9 ） 1626 - 1636 2006年05月

ISSN 0014-4827

Wnt1 and -5a have been shown to modulate the proliferation and differentiation of midbrain dopaminergic (DA) neurons. However, it is not known whether other Wnts or which Frizzled (Fz) receptors are expressed in the developing midbrain. We found that 13 out of 19 Wnts, all 10 Fzs, and several intracellular Wnt signaling modulators, including Axin, FRAT, Naked, Par-1, and Ltap are developmentally regulated between embryonic days (E) 10.5 and 15.5. Next, we studied whether Fzs are differentially expressed in different cell types and examined neuronal-progenitor- or glial-enriched cultures and DA neurons isolated from TH-GFP reporter mice. We found that Fz8 is expressed at high levels in DA neurons at E11.5 and E13.5. Fz6 and -7 are the predominant transcripts in glial precursors, and Fz9, which is absent in DA neurons at E11.5, is the main receptor expressed in neuronal precursors. We therefore examined the function of Fz9 in DA cells and found that overexpression of Fz9 reduced Wnt5a- but not Wnt3a-induced hyperphosphorylation of Dishevelled. Thus, our results show that Fzs are developmentally regulated and differentially expressed in VM precursors, DA neurons, and glia. These findings suggest that Fz expression contributes to provide specificity to Wnt-mediated effects. © 2006 Elsevier Inc. All rights reserved.

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The Sox2 regulatory region 2 functions as a neural stem cell-specific enhancer in the telencephalon

Miyagi Satoru, Nishimoto Masazumi, Saito Tetsuichiro, Ninomiya Mikiko, Sawamoto Kazunobu, Okano Hideyuki, Muramatsu Masami, Oguro Hideyuki, Iwama Atsushi, Okuda Akihiko

Journal of Biological Chemistry 281 （ 19 ） 13374 - 13381 2006年05月

ISSN 0021-9258

Sox2 is expressed at high levels in neuroepithelial stem cells and persists in neural stem/progenitor cells throughout adulthood. We showed previously that the Sox2 regulatory region 2 (SRR2) drives strong expression in these cells. Here we generated transgenic mouse strains with the β-geo reporter gene under the control of the SRR2 in order to examine the spatiotemporal function of this regulatory region. We show that the SRR2 functions specifically in neural stem/progenitor cells. However, unlike Nestin 2nd intronic enhancer, the SRR2 shows strong regional specificity functioning only in restricted areas of the telencephalon but not in any other portions of the central nervous system such as the spinal cord. We also show by in vitro clonogenic assay that at least some of these SRR2-functioning cells possess the hallmark properties of neural stem cells. In adult brains, we could detect strong β-geo expression in the subventricular zone of the lateral ventricle and along the rostral migrating stream where actively dividing cells reside. Chromatin immunoprecipitation assays reveal interactions of POU and Sox factors with SRR2 in neural stem/progenitor cells. Our data also suggest that the specific recruitment of these proteins to the SRR2 in the telencephalon defines the spatiotemporal activity of the enhancer in the developing nervous system. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

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A carbohydrate-binding protein, Galectin-1, promotes proliferation of adult neural stem cells

Sakaguchi Masanori, Shingo Tetsuro, Shimazaki Takuya, Okano Hirotaka James, Shiwa Mieko, Ishibashi Satoru, Oguro Hideyuki, Ninomiya Mikiko, Kadoya Toshihiko, Horie Hidenori, Shibuya Akira, Mizusawa Hidehiro, Poirier Françoise, Nakauchi Hiromitsu, Sawamoto Kazunobu, Okano Hideyuki

Proceedings of the National Academy of Sciences of the United States of America 103 （ 18 ） 7112 - 7117 2006年05月

ISSN 0027-8424

In the subventricular zone of the adult mammalian forebrain, neural stem cells (NSCs) reside and proliferate to generate young neurons. We screened factors that promoted the proliferation of NSCs in vitro by a recently developed proteomics technique, the ProteinChip system. In this screen, we identified a soluble carbohydrate-binding protein, Galectin-1, as a candidate. We show herein that Galectin-1 is expressed in a subset of slowly dividing subventricular zone astrocytes, which includes the NSCs. Based on results from intraventricular infusion experiments and phenotypic analyses of knockout mice, we demonstrate that Galectin-1 is an endogenous factor that promotes the proliferation of NSCs in the adult brain. © 2006 by The National Academy of Sciences of the USA.

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Novel Oncolytic HSV-1 Mutant Expressing ICP 34.5 under the Transcriptional Control of Musashi1 Promoter for the Treatment of Gliomas

岡野栄之

MOLECULAR THERAPY 13 S361 - S361 2006年05月

研究論文（学術雑誌）, ISSN 1525-0016

Role of Numb in dendritic spine development with a Cdc42 GEF intersectin and EphB2

Nishimura Takashi, Yamaguchi Tomoya, Tokunaga Akinori, Hara Akitoshi, Hamaguchi Tomonari, Kato Katsuhiro, Iwamatsu Akihiro, Okano Hideyuki, Kaibuchi Kozo

Molecular Biology of the Cell 17 （ 3 ） 1273 - 1285 2006年03月

ISSN 1059-1524

Numb has been implicated in cortical neurogenesis during nervous system development, as a result of its asymmetric partitioning and antagonizing Notch signaling. Recent studies have revealed that Numb functions in clathrin-dependent endocytosis by binding to the AP-2 complex. Numb is also expressed in postmitotic neurons and plays a role in axonal growth. However, the functions of Numb in later stages of neuronal development remain unknown. Here, we report that Numb specifically localizes to dendritic spines in cultured hippocampal neurons and is implicated in dendritic spine morphogenesis, partially through the direct interaction with intersectin, a Cdc42 guanine nucleotide exchange factor (GEF). Intersectin functions as a multidomain adaptor for proteins involved in endocytosis and cytoskeletal regulation. Numb enhanced the GEF activity of intersectin toward Cdc42 in vivo. Expression of Numb or intersectin caused the elongation of spine neck, whereas knockdown of Numb and Numb-like decreased the protrusion density and its length. Furthermore, Numb formed a complex with EphB2 receptor-type tyrosine kinase and NMDA-type glutamate receptors. Knockdown of Numb suppressed the ephrin-B1-induced spine development and maturation. These results highlight a role of Numb for dendritic spine development and synaptic functions with intersectin and EphB2. © 2006 by The American Society for Cell Biology.

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Proinflammatory cytokines expression in noise-induced damaged cochlea

Fujioka Masato, Kanzaki Sho, Okano Hirotaka James, Masuda Masatsugu, Ogawa Kaoru, Okano Hideyuki

Journal of Neuroscience Research 83 （ 4 ） 575 - 583 2006年03月

ISSN 0360-4012

Recent studies have showed that inflammatory responses occur in inner ear under various damaging conditions including noise-overstimulation. We evaluated the time-dependent expression of proinflammatory cytokines in noise-exposed rat cochlea. Among several detected cytokines, real-time RT-PCR showed that interleukin-1beta (IL-1β) and interleukin-6 (IL-6) were significantly induced 3 hr after noise exposure, and quickly downregulated to the basal level. Tumor necrosis factor-alpha (TNF-α) was also slightly upregulated immediately after noise exposure, Immunohistochemical analysis showed that IL-6 expression was distinctively induced within the lateral side of the spiral ligament. Sequential expression analysis showed that IL-6 immunoreactivity was initially found in the cytoplasm of lateral wall cells, including Type IV and III fibrocytes, and expanded broader throughout the lateral wall, finally to the stria vascularis. Because of the negative Iba-1 staining, IL-6 expression in the early-phase was not due to macrophage or microglia activation. IL-6 was also detected in spiral ganglion neurons at 12 and 24 hr after noise exposure. Our data demonstrates the production of proinflammatory cytokines, including TNF-α, IL-1β, and IL-6, in early phase of noise overstimulated cochlea. IL-6 expression was observed in the spiral ligament, stria vascularis, and spiral ganglion neurons. These cytokines, produced by the cochlear structure itself in response to noise exposure, may initiate an inflammatory response and have some role in the mechanism of noise-induced cochlear damage. © 2006 Wiley-Liss, Inc.

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Histopathological and behavioral improvement of murine mucopolysaccharidosis type VII by intracerebral transplantation of neural stem cells

Fukuhara Yasuyuki, Li Xiao Kang, Kitazawa Yusuke, Inagaki Masumi, Matsuoka Kentaro, Kosuga Motomichi, Kosaki Rika, Shimazaki Takuya, Endo Hitoshi, Umezawa Akihiro, Okano Hideyuki, Takahashi Takao, Okuyama Torayuki

Molecular Therapy 13 （ 3 ） 548 - 555 2006年03月

ISSN 1525-0016

The therapeutic efficacy of neural stem cell transplantation for central nervous system (CNS) lesions in lysosomal storage disorders was explored using a murine model of mucopolysaccharidosis type VII (MPS VII). We used fetal neural stem cells derived from embryonic mouse striata and expanded in vitro by neurosphere formation as the source of graft materials. We transplanted neurospheres into the lateral ventricles of newborn MPS VII mice and found that donor cells migrated far beyond the site of injection within 24 h, and some of them could reach the olfactory bulb. A quantitative measurement indicated that the GUSB activity in the brain was 12.5 to 42.3% and 5.5 to 6.3% of normal activity at 24 h and 3 weeks after transplantation. In addition, histological analysis revealed a widespread decrease in lysosomal storage in the recipient's hippocampus, cortex, and ependyma. A functional assessment with novel-object recognition tests confirmed improvements in behavioral patterns. These results suggest that intracerebral transplantation of neural stem cells is feasible for treatment of CNS lesions associated with lysosomal storage disorders. Copyright © The American Society of Gene Therapy.

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New neurons follow the flow of cerebrospinal fluid in the adult brain

Sawamoto Kazunobu, Wichterle Hynek, Gonzalez-Perez Oscar, Cholfin Jeremy A., Yamada Masayuki, Spassky Nathalie, Murcia Noel S., Garcia-Verdugo Jose Manuel, Marin Oscar, Rubenstein John L R, Tessier-Lavigne Marc, Okano Hideyuki, Alvarez-Buylla Arturo

Science 311 （ 5761 ） 629 - 632 2006年02月

ISSN 0036-8075

In the adult brain, neuroblasts born in the subventricular zone migrate from the walls of the lateral ventricles to the olfactory bulb. How do these cells orient over such a long distance and through complex territories? Here we show that neuroblast migration parallels cerebrospinal fluid (CSF) flow. Beating of ependymal cilia is required for normal CSF flow, concentration gradient formation of CSF guidance molecules, and directional migration of neuroblasts. Results suggest that polarized epithelial cells contribute important vectorial information for guidance of young, migrating neurons.

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Administration of hematopoietic cytokines in the subacute phase after cerebral infarction is effective for functional recovery facilitating proliferation of intrinsic neural stem/progenitor cells and transition of bone marrow-derived neuronal cells

Kawada Hiroshi, Takizawa Shunya, Takanashi Tomomi, Morita Yuko, Fujita Jun, Fukuda Keiichi, Takagi Shigeharu, Okano Hideyuki, Ando Kiyoshi, Hotta Tomomitsu

Circulation 113 （ 5 ） 701 - 710 2006年02月

ISSN 0009-7322

Background - Hematopoietic cytokines, granulocyte colony-stimulating factor (G-CSF), and stem cell factor (SCF) were reported to show a neuroprotective effect or to support neurogenesis. These cytokines also mobilize bone marrow (BM) cells into the brain, and the BM-derived cells differentiate into neuronal cells. We administered these hematopoietic cytokines after focal cerebral ischemia and assessed their effects and the therapeutic time window for neuronal regeneration. Methods and Results - We induced permanent middle cerebral artery occlusion in mice whose BM had been replaced with BM cells from green fluorescent protein (GFP)-transgenic mice. The occluded mice were treated with G-CSF and SCF in the acute phase (days 1 to 10) or subacute phase (days 11 to 20), and the brain functions and histological changes were evaluated. Separately, we injected bromodeoxyuridine during cytokine treatment to assess cell kinetics in the brain. Six mice were prepared for each experimental group. Administration of G-CSF and SCF in the subacute phase effectively improved not only motor performance but also higher brain function, compared with acute-phase treatment. Acute-phase and subacute-phase treatments identically reduced the infarct volume relative to vehicle treatment. However, subacute-phase treatment significantly induced transition of BM-derived neuronal cells into the peri-infarct area and stimulated proliferation of intrinsic neural stem/progenitor cells in the neuroproliferative zone. Conclusions - Administration of G-CSF and SCF in the subacute phase after focal cerebral ischemia is effective for functional recovery, enhancing cytokine-induced generation of neuronal cells from both BM-derived cells and intrinsic neural stem/progenitor cells. Because G-CSF and SCF are available for clinical use, these findings suggest a new therapeutic strategy for stroke. © 2006 American Heart Association, Inc.

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Distinct functions of human numb isoforms revealed by misexpression in the neural stem cell lineage in the Drosophila larval brain

Toriya Masako, Tokunaga Akinori, Sawamoto Kazunobu, Nakao Keiko, Okano Hideyuki

Developmental Neuroscience 28 （ 1-2 ） 142 - 155 2006年02月

ISSN 0378-5866

Mammalian Numb (mNumb) has multiple functions and plays important roles in the regulation of neural development, including maintenance of neural progenitor cells and promotion of neuronal differentiation in the central nervous system (CNS). However, the molecular bases underlying the distinct functions of Numb have not yet been elucidated. mNumb, which has four splicing isoforms, can be divided into two types based on the presence or absence of an amino acid insert in the proline-rich region (PRR) in the C-terminus. It has been proposed that the distinct functions of mNumb may be attributable to these two different types of isoforms. In this study, we used the outer optic anlage (OOA) of the Drosophila larval brain as an assay system to analyze the functions of these two types of isoforms in the neural stem cells, since the proliferation pattern of neuroepithelial (NE) stem cells in the OOA closely resembles that of the vertebrate neural stem/progenitor cells. They divide to expand the progenitor cell pool during early neurogenesis and to produce neural precursors/neurons during late neurogenesis. Clonal analysis in the OOA allows one to discriminate between the NE stem cells, which divide symmetrically to expand the progenitor pool, and the postembryonic neuroblasts (pNBs), which divide asymmetrically to produce neural precursors (ganglion mother cells), each of which divides once to produce two neurons. We found that in the OOA, the human Numb isoform with a long PRR domain (hNumb-PRRL), which is mainly expressed during early neurogenesis in the mouse CNS, promotes proliferation of both NE cells and pNBs without affecting neuronal differentiation, while the other type of hNumb isoform with a short PRR domain (hNumb-PRRS), which is expressed throughout neurogenesis in the mouse embryonic CNS, inhibits proliferation of the stem cells and promotes neuronal differentiation. We also found that hNumb-PRRS, a functional homologue of Drosophila Numb, more strongly decreases the amount of nuclear Notch than hNumb-PRRL, and could antagonize Notch functions probably through endocytic degradation, suggesting that the two distinct types of hNumb isoforms could contribute to different phases of neurogenesis in the mouse embryonic CNS. Copyright © 2006 S. Karger AG.

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Lecithinized superoxide dismutase (PC-SOD) improved spinal cord injury-induced motor dysfunction through suppression of oxidative stress and enhancement of neurotrophic factor production

Takenaga Mitsuko, Ohta Yuki, Tokura Yukie, Hamaguchi Akemi, Nakamura Masaya, Okano Hideyuki, Igarashi Rie

Journal of Controlled Release 110 （ 2 ） 283 - 289 2006年01月

ISSN 0168-3659

PC-SOD (lecithinized superoxide dismutase) is a derivative of human Cu, Zn-SOD conjugated with 4 molecules of lecithin, yet having the enzyme activity of scavenging superoxide anion (O₂^-). Intravenous administration of PC-SOD promoted the recovery from spinal cord injury (SCI)-induced motor dysfunction in a dose-dependent manner in rat model, when evaluated by BBB (Basso Beattie Bresnahan) score. Even when given at 24 h after SCI, PC-SOD (1 mg/kg) significantly improved motor dysfunction. Distribution study demonstrated that PC-SOD gradually accumulated to the injured site. Enzyme-linked immunoassay revealed that PC-SOD prevented quantitative loss of neurons, astrocytes, and oligodendrocytes. PC-SOD inhibited SCI-induced oxidative stress, such as the decrease of free sulfhydryl residue, acetylcholine esterase activity, and the increase of lipid peroxidation. PC-SOD increased the production of neuroprotective factors. HIF-1α gene expression increased following SCI, and PC-SOD further increased it. In conclusion, PC-SOD gradually accumulated and retained at the damaged site to scavenge excessive O ₂^-, and suppressed neuronal death through reducing oxidative stress, increasing neuroprotective factor production and HIF-1α gene expression. © 2005 Elsevier B.V. All rights reserved.

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Expression of Musashi-1 in the rat stomach and changes during mucosal injury and restitution

Nagata Hiroshi, Akiba Yasutada, Suzuki Hidekazu, Okano Hideyuki, Hibi Toshifumi

FEBS Letters 580 （ 1 ） 27 - 33 2006年01月

ISSN 0014-5793

Musashi-1 is involved in maintenance of neural stem cells. In the rat stomach, we found some cells in the luminal compartment of the mucosa that stained positively for Musashi-1. These cells were distinct from other cells of epithelial lineage, except for parietal cells, and coexpressed HES5. The Musashi-1-positive cells exfoliated after damage, while Musashi-1 expression in neck cells were upregulated, and proliferating cells diminished before reappearing and increasing in number thereafter. We conclude that a subpopulation of parietal cells acts as a source of Musashi-1, which contributes to rapid re-epithelization by restoration of stem cells and regulation of cell differentiation. © 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

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Disease progression of human SOD1 (G93A) transgenic ALS model rats

Matsumoto Arifumi, Okada Yohei, Nakamichi Masanori, Nakamura Masaya, Toyama Yoshiaki, Sobue Gen, Nagai Makiko, Aoki Masashi, Itoyama Yasuto, Okano Hideyuki

Journal of Neuroscience Research 83 （ 1 ） 119 - 133 2006年01月

ISSN 0360-4012

The recent development of a rat model of amyotrophic lateral sclerosis (ALS) in which the rats harbor a mutated human SOD1 (G93A) gene has greatly expanded the range of potential experiments, because the rats' large size permits biochemical analyses and therapeutic trials, such as the intrathecal injection of new drugs and stem cell transplantation. The precise nature of this disease model remains unclear. We described three disease phenotypes: the forelimb-, hindlimb-, and general-types. We also established a simple, non-invasive, and objective evaluation system using the body weight, inclined plane test, cage activity, automated motion analysis system (SCANET), and righting reflex. Moreover, we created a novel scale, the Motor score, which can be used with any phenotype and does not require special apparatuses. With these methods, we uniformly and quantitatively assessed the onset, progression, and disease duration, and clearly presented the variable clinical course of this model; disease progression after the onset was more aggressive in the forelimb-type than in the hindlimb-type. More importantly, the disease stages defined by our evaluation system correlated well with the loss of spinal motor neurons. In particular, the onset of muscle weakness coincided with the loss of approximately 50% of spinal motor neurons. This study should provide a valuable tool for future experiments to test potential ALS therapies. © 2005 Wiley-Liss, Inc.

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Expression of RNA-binding protein Musashi in hair follicle development and hair cycle progression

Sugiyama-Nakagiri Yoriko, Akiyama Masashi, Shibata Shinsuke, Okano Hideyuki, Shimizu Hiroshi

American Journal of Pathology 168 （ 1 ） 80 - 92 2006年01月

ISSN 0002-9440

Epithelial stem cells reside in the hair follicle (HF) bulge region and possess the ability to differentiate into a variety of cutaneous epithelial cells. The evolutionarily conserved Musashi family of RNA-binding proteins is associated with maintenance and/or asymmetric cell division of neural progenitor cells, and a mammalian Musashi protein is expressed in various epithelial stem/progenitor cells, including gut, stomach, and mammary gland. Thus, we hypothesized that Musashi might be expressed to stem cells and early progenitor cells of HF epithelium. Reverse transcriptase-polymerase chain reaction and immunoblotting identified Musashi-1 (Msl-1) and Musashi-2 (Msi-2) mRNA and protein in cultured mouse keratinocytes, but only Msi-1 was identified in human keratinocytes. In mice, immunohistochemical studies showed that Msi-1 and Msi-2 were expressed in the epidermis and HFs from E14.5 until adulthood. In the early anagen phase, Msi-1 and Msi-2 were expressed in the bulge and secondary germ cells and subsequently in inner root sheath (IRS) cells, especially the middle IRS cells, during the late anagen phase. In human skin, MsM was detected in fetal HF cells but not in adult HFs. These observations suggest that Musashi functions not only in the asymmetric division of early progenitor cells but also in the differentiation of IKS cells during HF development and hair cycle progression. Copyright © American Society for Investigative Pathology.

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The role of autophagy in intracellular quality control

岡野栄之

AUTOPHAGY 2 （ 4 ） 355 - 355 2006年01月

研究論文（学術雑誌）, ISSN 1554-8627

Enhanced therapeutic efficacy of G207 for the treatment of glioma through Musashi1 promoter retargeting of γ34.5-mediated virulence

Kanai R., Tomita H., Shinoda A., Takahashi M., Goldman S., Okano H., Kawase T., Yazaki T.

Gene Therapy 13 （ 2 ） 106 - 116 2006年01月

ISSN 0969-7128

G207 is a conditionally replicating derivative of herpes simplex virus type1 (HSV-1) engineered with deletions of both ICP34.5 loci and a lacZ insertion disabling the ICP6 gene. G207 exhibits an efficient oncolytic activity in vitro and in vivo, yet minimal toxicity in normal tissue, and is now in clinical trial for malignant glioma. According to the results of clinical trials, however, although G207 was proved to be safe, the efficacy was not so impressive. Deletion of the ICP34.5 gene coding for virulence made G207 extremely safe, but it markedly reduced the cytotoxicity mediated by HSV-1. To enhance the therapeutic efficacy of G207 without diminishing its safety, we used a defective vector containing Musashi1 promoter/ICP34.5, with G207 as helper virus. P/ musashi1 was functional selectively in human glioma cell lines (U87MG, U251, T98G) in this study and dvM345 showed a much higher therapeutic efficacy both in culture and in the in vivo glioma model, than G207 alone, without diminishing its favorable toxicity profile. These results suggest that transcriptional regulation of ICP34.5 by P/musashi1 can be used to target HSV-1 virulence toward gliomas while maintaining the desirable neuroattenuated phenotype. © 2006 Nature Publishing Group All rights reserved.

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Subventricular zone-derived neuroblasts migrate and differentiate into mature neurons in the post-stroke adult striatum

Yamashita Toru, Ninomiya Mikiko, Acosta Pilar Hernández, García-Verdugo Jose Manuel, Sunabori Takehiko, Sakaguchi Masanori, Adachi Kazuhide, Kojima Takuro, Hirota Yuki, Kawase Takeshi, Araki Nobuo, Abe Koji, Okano Hideyuki, Sawamoto Kazunobu

Journal of Neuroscience 26 （ 24 ） 6627 - 6636 2006年

ISSN 0270-6474

Recent studies have revealed that the adult mammalian brain has the capacity to regenerate some neurons after various insults. However, the precise mechanism of insult-induced neurogenesis has not been demonstrated. In the normal brain, GFAP-expressing cells in the subventricular zone (SVZ) of the lateral ventricles include a neurogenic cell population that gives rise to olfactory bulb neurons only. Herein, we report evidence that, after a stroke, these cells are capable of producing new neurons outside the olfactory bulbs. SVZ GFAP-expressing cells labeled by a cell-type-specific viral infection method were found to generate neuroblasts that migrated toward the injured striatum after middle cerebral artery occlusion. These neuroblasts in the striatum formed elongated chain-like cell aggregates similar to those in the normal SVZ, and these chains were observed to be closely associated with thin astrocytic processes and blood vessels. Finally, long-term tracing of the green fluorescent-labeled cells with a Cre-loxP system revealed that the SVZ-derived neuroblasts differentiated into mature neurons in the striatum, in which they expressed neuronal-specific nuclear protein and formed synapses with neighboring striatal cells. These results highlight the role of the SVZ in neuronal regeneration after a stroke and its potential as an important therapeutic target for various neurological disorders. Copyright © 2006 Society for Neuroscience.

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In vivo and in vitro analysis of Musashi family in mammalian CNS and PNS development

岡野栄之

NEUROSCIENCE RESEARCH 55 S182 - S182 2006年

研究論文（学術雑誌）, ISSN 0168-0102

Therapeutic potential of CD8 alpha(+) dendritic cells (DCs) for the treatment of mouse spinal cord injury

岡野栄之

NEUROSCIENCE RESEARCH 55 S217 - S217 2006年

研究論文（学術雑誌）, ISSN 0168-0102

Implication of "Down syndrome cell adhesion molecule" in the hippocampal neurogenesis of ischemic monkeys

岡野栄之

HIPPOCAMPUS 16 （ 11 ） 924 - 935 2006年

研究論文（学術雑誌）, ISSN 1050-9631

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HGF gene therapy for the treatment of spinal cord injury

岡野栄之

NEUROSCIENCE RESEARCH 55 S13 - S13 2006年

研究論文（学術雑誌）, ISSN 0168-0102

Role of beta-catenin signaling in regulating proliferation of transit-amplifying cells in the adult mouse subventricular zone

岡野栄之

NEUROSCIENCE RESEARCH 55 S53 - S53 2006年

研究論文（学術雑誌）, ISSN 0168-0102

A role of neuronal RNA binding protein Hu and NF45/90 in the nervous system

岡野栄之

NEUROSCIENCE RESEARCH 55 S145 - S145 2006年

研究論文（学術雑誌）, ISSN 0168-0102

cdk5 is required for neuroblast migration in the adult mouse brain

岡野栄之

NEUROSCIENCE RESEARCH 55 S52 - S52 2006年

研究論文（学術雑誌）, ISSN 0168-0102

Effects of stromal cell secretory factors on ES cell-derived neural stem/progenitor cells

岡野栄之

NEUROSCIENCE RESEARCH 55 S241 - S241 2006年

研究論文（学術雑誌）, ISSN 0168-0102

Identification of a neural specific gene by microarray and EST database analysis

岡野栄之

NEUROSCIENCE RESEARCH 55 S104 - S104 2006年

研究論文（学術雑誌）, ISSN 0168-0102

Cell surface complex type N-glycans are distinctive markers for prospective purification of neural stem cells

岡野栄之

NEUROSCIENCE RESEARCH 55 S105 - S105 2006年

研究論文（学術雑誌）, ISSN 0168-0102

Targeted gene transfer of human Hepatocyte Growth Factor into rat skin by the use of Iaser-induced stress waves

Terakawa Mitsuhiro, Sato Shunichi, Saitoh Daizoh, Ashida Hiroshi, Okano Hideyuki, Obara Minoru

2006年

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Transcription factor protein expression patterns by neural or neuronal progenitor cells of adult monkey subventricular zone

Tonchev A. B., Yamashima T., Sawamoto K., Okano H.

Neuroscience 139 （ 4 ） 1355 - 1367 2006年

ISSN 0306-4522

The anterior subventricular zone of the adult mammalian brain contains progenitor cells which are upregulated after cerebral ischemia. We have previously reported that while a part of the progenitors residing in adult monkey anterior subventricular zone travels to the olfactory bulb, many of these cells sustain location in the anterior subventricular zone for months after injury, exhibiting a phenotype of either neural or neuronal precursors. Here we show that ischemia increased the numbers of anterior subventricular zone progenitor cells expressing developmentally regulated transcription factors including Pax6 (paired-box 6), Emx2 (empty spiracles-homeobox 2), Sox 1-3 (sex determining region Y-box 1-3), Ngn1 (neurogenin 1), Dlx1,5 (distalless-homeobox 1,5), Olig1,3 (oligodendrocyte lineage gene 1,3) and Nkx2.2 (Nk-box 2.2), as compared with control brains. Analysis of transcription factor protein expression by sustained neural or neuronal precursors in anterior subventricular zone revealed that these two cell types were positive for characteristic sets of transcription factors. The proteins Pax6, Emx2, Sox2,3 and Olig1 were predominantly localized to dividing neural precursors while the factors Sox1, Ngn1, Dlx1,5, Olig2 and Nkx2.2 were mainly expressed by neuronal precursors. Further, differences between monkeys and non-primate mammals emerged, related to expression patterns of Pax6, Olig2 and Dlx2. Our results suggest that a complex network of developmental signals might be involved in the specification of primate progenitor cells. © 2006 IBRO.

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Neuroprotection and neurosupplementation in ischaemic brain

Yamashita T., Deguchi K., Sawamoto K., Okano H., Kamiya T., Abe K.

Biochemical Society Transactions 34 （ 6 ） 1310 - 1312 2006年

ISSN 0300-5127

Possible strategies for treating ischaemic stroke include: (i) neuroprotection (preventing damaged neurons from undergoing apoptosis in the acute phase of cerebral ischaemia), and (ii) neurosupplementation (the repair of broken neuronal networks with newly born neurons in the chronic phase of cerebral ischaemia). In this paper, we review our recent progress in development of these distinct new strategies for treatment of damaged brain following a stroke. Firstly, we investigated the role of endogenous IL-6 (interleukin-6), which is one of the cytokines drastically induced by ischaemic stimuli, by administering IL-6RA (anti-IL-6 receptor monoclonal antibody) to mice.We found that endogenous IL-6 plays a critical role in neuroprotection and that its role may be mediated by STAT3 (signal transducer and activator of transcription-3) activation. Secondly, we studied the endogenous sources of the newly born neurons in the ischaemic striatum by region- and cell-type-specific cell labelling techniques. The results revealed that the SVZ (subventricular zone) is the principal source of the neuronal progenitors that migrate laterally towards the infarcted regions, and differentiate into newly born neurons. Finally, we developed a restorative stroke therapy with a bio-affinitive scaffold, which is an appropriate poly-porous structure releasing bioactive substances such as neurotrophic factor. This bio-affinitive scaffold is able to give an appropriate environment for newly born neurons. In future, we will combine these strategies to develop more effective therapies for treatment of strokes. ©2006 Biochemical Society.

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Separation of μ-opioid receptor desensitization and internalization

Arttamangkul Seksiri, Torrecilla Maria, Kobayashi Kazuto, Okano Hideyuki, Williams John T.

Journal of Neuroscience 26 （ 15 ） 4118 - 4125 2006年

ISSN 0270-6474

A close relationship between desensitization and internalization of μ-opioid receptors (MORs) has been proposed based on differential actions of series of agonists. The role that these two processes have in the development of tolerance and dependence to opioids has been a controversial subject that has been studied in a variety of model systems. Here, we examine desensitization and internalization of endogenous MORs simultaneously in primary cultures of locus ceruleus neurons using fluorescently tagged peptide agonists. With the use of two fluorescent opioid peptides, dermorphin-Bodipy Texas Red and dermorphin-Alexa594 (Derm-A594), desensitization was measured electrophysiologically and trafficking was followed by the accumulation of intracellular fluorescent puncta. Blocking endocytosis with concanavalin A eliminated the accumulation of fluorescent puncta but desensitization induced by Derm-A594 was unaffected. Likewise, after treatment with concanavalin A, there was no change in either desensitization or recovery from desensitization induced by [Met]⁵ enkephalin. The results demonstrate that desensitization and the recovery from desensitization are not dependent on receptor internalization and suggest that the activity of endogenous MORs in primary neurons can be modulated at the level of the plasma membrane. Copyright © 2006 Society for Neuroscience.

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Adult neural stem cells and central nervous system repair.

Okano H.

Ernst Schering Research Foundation workshop （ 60 ） 215 - 228 2006年

ISSN 0947-6075

It has long been believed that the adult mammalian central nervous system does not regenerate after injury. However, recent advances in the field of stem cell biology, including the identification of Musashi-1-positive neural stem cells (NSCs) or NSC-like cells, has provided new insight for the development of novel therapeutic strategies aimed at inducing regeneration in the damaged central nervous system (CNS). The major strategies for inducing regeneration in the damaged CNS can be classified into two subgroups: (1) activation of endogenous neural stem cells and (2) cell transplantation therapies. In this paper, we would like to summarize our recent findings on the functions of the neural RNA-binding protein Musashi-l expressed in neural stem cells in relation to insult-induced neurogenesis, and therapeutic interventions for spinal cord injury, especially focusing on the treatment of spinal cord injury in the acute phase with anti-IL-6 receptor blocking antibody.

Gene transfer of human hepatocyte growth factor by the use of nanosecond pulsed laser-induced stress waves

Terakawa Mitsuhiro, Sato Shunichi, Saitoh Daizoh, Hasegawa Makoto, Ashida Hiroshi, Okano Hideyuki, Obara Minoru

2006年

We successfully delivered a therapeutic vector construct, which carries hepatocyte growth factor (HGF) gene, to rat skin in vivo. After HGF expression vector had been intradermally injected to rat skin, LISWs were generated by irradiating the laser target put on the rat skin with nanosecond pulses from the second harmonics (532 nm) of a Q-switched Nd:YAG laser. Concentration of HGF protein increased by a factor of four by the application of LISWs when compared with that of control samples without LISW application. We also investigated the effects of LISWs on the integrity of plasmid DNA.

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Laser-induced stress wave-assisted gene transfection with cationic liposome

岡野栄之

JAPANESE JOURNAL OF APPLIED PHYSICS PART 2-LETTERS & EXPRESS LETTERS 46 （ 45-49 ） L1243 - L1245 2007年12月

研究論文（学術雑誌）, ISSN 0021-4922

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Coordinated expression of HuD and GAP-43 in hippocampal dentate granule cells during developmental and adult plasticity

岡野栄之

NEUROCHEMICAL RESEARCH 32 （ 12 ） 2142 - 2151 2007年12月

研究論文（学術雑誌）, ISSN 0364-3190

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Laser-induced stress wave-assisted gene transfection with cationic liposome

岡野栄之

JAPANESE JOURNAL OF APPLIED PHYSICS PART 2-LETTERS & EXPRESS LETTERS 46 （ 45-49 ） L1243 - L1245 2007年12月

研究論文（学術雑誌）, ISSN 0021-4922

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Identification of a neuron-specific human gene, KIAA1110, that is a guanine nucleotide exchange factor for ARF1

岡野栄之

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 364 （ 4 ） 737 - 742 2007年12月

研究論文（学術雑誌）, ISSN 0006-291X

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RMD-1, a novel microtubule-associated protein, functions in chromosome segregation in Caenorhabditis elegans

岡野栄之

JOURNAL OF CELL BIOLOGY 179 （ 6 ） 1149 - 1162 2007年12月

研究論文（学術雑誌）, ISSN 0021-9525

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Visceral afferents directly activate catecholamine neurons in the solitary tract nucleus

岡野栄之

JOURNAL OF NEUROSCIENCE 27 （ 48 ） 13292 - 13302 2007年11月

研究論文（学術雑誌）, ISSN 0270-6474

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Visceral afferents directly activate catecholamine neurons in the solitary tract nucleus

岡野栄之

JOURNAL OF NEUROSCIENCE 27 （ 48 ） 13292 - 13302 2007年11月

研究論文（学術雑誌）, ISSN 0270-6474

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Use of an epidural cooling catheter with a closed countercurrent lumen to protect against ischemic spinal cord injury in pigs

岡野栄之

JOURNAL OF THORACIC AND CARDIOVASCULAR SURGERY 134 （ 5 ） 1220 - 1226 2007年11月

研究論文（学術雑誌）, ISSN 0022-5223

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beta-Catenin signaling promotes proliferation of progenitor cells in the adult mouse subventricular zone

岡野栄之

STEM CELLS 25 （ 11 ） 2827 - 2836 2007年11月

研究論文（学術雑誌）, ISSN 1066-5099

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Cyclin-dependent kinase 5 is required for control of neuroblast migration in the postnatal subventricular zone

岡野栄之

JOURNAL OF NEUROSCIENCE 27 （ 47 ） 12829 - 12838 2007年11月

研究論文（学術雑誌）, ISSN 0270-6474

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Angiotensin II type I receptor blockade prevents decrease in adult stem-like cells in kidney after ureteral obstruction

岡野栄之

EUROPEAN JOURNAL OF PHARMACOLOGY 573 （ 1-3 ） 216 - 220 2007年11月

研究論文（学術雑誌）, ISSN 0014-2999

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Side population in human uterine myometrium displays phenotypic and functional characteristics of myometrial stem cells

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 104 （ 47 ） 18700 - 18705 2007年11月

研究論文（学術雑誌）, ISSN 0027-8424

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Administration of granulocyte colony-stimulating factor after myocardial infarction enhances the recruitment of hematopoietic stem cell-derived myofibroblasts and contributes to cardiac repair

岡野栄之

STEM CELLS 25 （ 11 ） 2750 - 2759 2007年11月

研究論文（学術雑誌）, ISSN 1066-5099

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In vivo tracing of neural tracts in the intact and injured spinal cord of marmosets by diffusion tensor tractography

岡野栄之

JOURNAL OF NEUROSCIENCE 27 （ 44 ） 11991 - 11998 2007年10月

研究論文（学術雑誌）, ISSN 0270-6474

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Galectin-1 regulates neurogenesis in the subventricular zone and promotes functional recovery after stroke

岡野栄之

EXPERIMENTAL NEUROLOGY 207 （ 2 ） 302 - 313 2007年10月

研究論文（学術雑誌）, ISSN 0014-4886

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Regeneration of the central nervous system using endogenous repair mechanisms

岡野栄之

JOURNAL OF NEUROCHEMISTRY 102 （ 5 ） 1459 - 1465 2007年09月

研究論文（学術雑誌）, ISSN 0022-3042

Effects of 120 mT static magnetic field on TGF-beta 1-inhibited endothelial tubular formation in vitro

岡野栄之

BIOELECTROMAGNETICS 28 （ 6 ） 497 - 499 2007年09月

研究論文（学術雑誌）, ISSN 0197-8462

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Effects of 12 mT static magnetic field on sympathetic agonist-induced hypertension in Wistar rats

岡野栄之

BIOELECTROMAGNETICS 28 （ 5 ） 369 - 378 2007年07月

研究論文（学術雑誌）, ISSN 0197-8462

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Epidermal growth factor signaling mediated by Grb2 associated binder1 is required for the spatiotemporally regulated proliferation of Olig2-expressing progenitors in the embryonic spinal cord

岡野栄之

STEM CELLS 25 （ 6 ） 1410 - 1422 2007年06月

研究論文（学術雑誌）, ISSN 1066-5099

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Epidermal growth factor signaling mediated by Grb2 associated binder1 is required for the spatiotemporally regulated proliferation of Olig2-expressing progenitors in the embryonic spinal cord

岡野栄之

STEM CELLS 25 （ 6 ） 1410 - 1422 2007年06月

研究論文（学術雑誌）, ISSN 1066-5099

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Interleukin-6 receptor-mediated activation of signal transducer and activator of transcription-3 (STAT3) promotes choroidal neovascularization

岡野栄之

AMERICAN JOURNAL OF PATHOLOGY 170 （ 6 ） 2149 - 2158 2007年06月

研究論文（学術雑誌）, ISSN 0002-9440

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Neural stem cells: Their involvement in adult neurogenesis and CNS-repair

岡野栄之

INFLAMMATION RESEARCH 56 S347 - S348 2007年06月

研究論文（学術雑誌）, ISSN 1023-3830

Impaired motor functions in mice lacking the RNA-binding protein Hzf

岡野栄之

NEUROSCIENCE RESEARCH 58 （ 2 ） 183 - 189 2007年06月

研究論文（学術雑誌）, ISSN 0168-0102

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Musashi-I expression in postnatal mouse olfactory epithelium

岡野栄之

NEUROREPORT 18 （ 7 ） 641 - 644 2007年05月

研究論文（学術雑誌）, ISSN 0959-4965

Leptin suppresses ghrelin-induced activation of neuropeptide y neurons in the arcuate nucleus via phosphatidylinositol 3-kinase- and phosphodiesterase 3-mediated pathway

岡野栄之

ENDOCRINOLOGY 148 （ 5 ） 2251 - 2263 2007年05月

研究論文（学術雑誌）, ISSN 0013-7227

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Delayed gastric mucosal repair after ethanol intoxication in Musashi-1 null mice

岡野栄之

GASTROENTEROLOGY 132 （ 4 ） A270 - A271 2007年04月

研究論文（学術雑誌）, ISSN 0016-5085

beta-catenin activation and epithelial-mesenchymal transition in the pathogenesis of pterygium

岡野栄之

INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE 48 （ 4 ） 1511 - 1517 2007年04月

研究論文（学術雑誌）, ISSN 0146-0404

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Electrical stimulation modulates fate determination of differentiating embryonic stem cells

岡野栄之

STEM CELLS 25 （ 3 ） 562 - 570 2007年03月

研究論文（学術雑誌）, ISSN 1066-5099

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SOCS3 is required to temporally fine-tune photoreceptor cell differentiation

岡野栄之

DEVELOPMENTAL BIOLOGY 303 （ 2 ） 591 - 600 2007年03月

研究論文（学術雑誌）, ISSN 0012-1606

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Noninvasive and real-time assessment of reconstructed functional human endometrium in NOD/SCID/gamma(null)(c) immunodeficient mice

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 104 （ 6 ） 1925 - 1930 2007年02月

研究論文（学術雑誌）, ISSN 0027-8424

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Bone marrow-derived cells express matrix metalloproteinases and contribute to regression of liver fibrosis in mice

岡野栄之

HEPATOLOGY 45 （ 1 ） 213 - 222 2007年01月

研究論文（学術雑誌）, ISSN 0270-9139

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Augmented therapeutic efficacy of an oncolytic herpes simplex virus type 1 mutant expressing ICP34.5 under the transcriptional control of musashil promoter in the treatment of malignant glioma

岡野栄之

HUMAN GENE THERAPY 18 （ 1 ） 63 - 73 2007年01月

研究論文（学術雑誌）, ISSN 1043-0342

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Enhanced angiogenesis in grafted skins by laser-induced stress wave-assisted gene-transfer of hepatocyte growth factor

岡野栄之

JOURNAL OF BIOMEDICAL OPTICS 12 （ 3 ） 1046 - 1052 2007年01月

研究論文（学術雑誌）, ISSN 1083-3668

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Regeneration-based therapies for spinal cord injuries

岡野栄之

NEUROCHEMISTRY INTERNATIONAL 51 （ 2-4 ） 68 - 73 2007年01月

研究論文（学術雑誌）, ISSN 0197-0186

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An efficient gene delivery system into cells using a nano-size silicon needle

岡野栄之

NEUROSCIENCE RESEARCH 58 S242 - S242 2007年

研究論文（学術雑誌）, ISSN 0168-0102

Plasma as a scaffold for regeneration of neural precursor cells after transplantation into rats with spinal cord injury

岡野栄之

CELL TRANSPLANTATION 16 （ 1 ） 57 - 65 2007年

研究論文（学術雑誌）, ISSN 0963-6897

Inhibition of histone deacetylase activates side population cells in kidney and partially reverses chronic renal injury

岡野栄之

STEM CELLS 25 （ 10 ） 2469 - 2475 2007年

研究論文（学術雑誌）, ISSN 1066-5099

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Signals regulating the differentiation vs. maintenance of neural progenitor cells

岡野栄之

NEUROSCIENCE RESEARCH 58 S19 - S19 2007年

研究論文（学術雑誌）, ISSN 0168-0102

Two distinct stem cell lineages in murine bone marrow

岡野栄之

STEM CELLS 25 （ 5 ） 1213 - 1221 2007年

研究論文（学術雑誌）, ISSN 1066-5099

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Novel regulatory mechanism of Hes1 gene expression

岡野栄之

NEUROSCIENCE RESEARCH 58 S10 - S10 2007年

研究論文（学術雑誌）, ISSN 0168-0102

Epigenetic mechanism regulating fate specification and plasticity of neural cells

岡野栄之

NEUROSCIENCE RESEARCH 58 S19 - S19 2007年

研究論文（学術雑誌）, ISSN 0168-0102

Sox6 is expressed in mouse embryonic neural stem/progenitor cells (NSPCs) and contributes to their maintenance and/or survival

岡野栄之

NEUROSCIENCE RESEARCH 58 S199 - S199 2007年

研究論文（学術雑誌）, ISSN 0168-0102

Establishment of three-dimensional culture of neural stem/progenitor cells in collagen Type-1 Gel

岡野栄之

RESTORATIVE NEUROLOGY AND NEUROSCIENCE 25 （ 2 ） 109 - 117 2007年

研究論文（学術雑誌）, ISSN 0922-6028

Expression of the neural RNA-binding protein Musashi1 in pediatric brain tumors

岡野栄之

PEDIATRIC NEUROSURGERY 43 （ 4 ） 279 - 284 2007年

研究論文（学術雑誌）, ISSN 1016-2291

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A dual role for reactive astrocytes after spinal cord injury

岡野栄之

NEUROSCIENCE RESEARCH 58 S29 - S29 2007年

研究論文（学術雑誌）, ISSN 0168-0102

Plasma as a scaffold for regeneration of neural precursor cells after transplantation into rats with spinal cord injury

岡野栄之

CELL TRANSPLANTATION 16 （ 1 ） 57 - 65 2007年

研究論文（学術雑誌）, ISSN 0963-6897

Diffusion-Tensor Neuronal Fiber Tractography and Manganese-enhanced MR Imaging of Primate Visual Pathway in the Common Marmoset: Preliminary Results

岡野栄之

RADIOLOGY 249 （ 3 ） 855 - 864 2008年12月

研究論文（学術雑誌）, ISSN 0033-8419

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The Influence of a Gradient Static Magnetic Field on an Unstirred Belousov-Zhabotinsky Reaction

岡野栄之

BIOELECTROMAGNETICS 29 （ 8 ） 598 - 604 2008年12月

研究論文（学術雑誌）, ISSN 0197-8462

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Epigenetic regulation of neural cell differentiation plasticity in the adult mammalian brain

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 105 （ 46 ） 18012 - 18017 2008年11月

研究論文（学術雑誌）, ISSN 0027-8424

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Requirement for COUP-TFI and II in the temporal specification of neural stem cells in CNS development

岡野栄之

NATURE NEUROSCIENCE 11 （ 9 ） 1014 - 1023 2008年09月

研究論文（学術雑誌）, ISSN 1097-6256

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Roles of STAT3/SOCS3 pathway in regulating the visual function and ubiquitin-proteasome-dependent degradation of rhodopsin during retinal inflammation

岡野栄之

JOURNAL OF BIOLOGICAL CHEMISTRY 283 （ 36 ） 24561 - 24570 2008年09月

研究論文（学術雑誌）, ISSN 0021-9258

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Angiotensin II type 1 receptor signaling contributes to synaptophysin degradation and neuronal dysfunction in the diabetic retina

岡野栄之

DIABETES 57 （ 8 ） 2191 - 2198 2008年08月

研究論文（学術雑誌）, ISSN 0012-1797

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Differentiation of a murine intestinal epithelial cell line (MIE) toward the M cell lineage

岡野栄之

AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY 295 （ 2 ） G273 - G284 2008年08月

研究論文（学術雑誌）, ISSN 0193-1857

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Consequence of the loss of Sox2 in the developing brain of the mouse

岡野栄之

FEBS LETTERS 582 （ 18 ） 2811 - 2815 2008年08月

研究論文（学術雑誌）, ISSN 0014-5793

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Neurologic phenotype of schimke immuno-osseous dysplasia and neurodevelopmental expression of SMARCAL1

Deguchi Kimiko, Clewing Johanna M., Elizondo Leah I., Hirano Ryuki, Huang Cheng, Choi Kunho, Sloan Emily A., Lücke Thomas, Marwedel Katja M., Powell Ralph D., Santa Cruz Karen, Willaime-Morawek Sandrine, Inoue Ken, Lou Shu, Northrop Jennifer L., Kanemura Yonehiro, Van Der Kooy Derek, Okano Hideyuki, Armstrong Dawna L., Boerkoel Cornelius F.

Journal of Neuropathology and Experimental Neurology 67 （ 6 ） 565 - 577 2008年06月

ISSN 0022-3069

Schimke immuno-osseous dysplasia (OMIM 242900) is an uncommon autosomal-recessive multisystem disease caused by mutations in SMARCAL1 (swi/snf-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a-like 1), a gene encoding a putative chromatin remodeling protein. Neurologic manifestations identified to date relate to enhanced atherosclerosis and cerebrovascular disease. Based on a clinical survey, we determined that half of Schimke immuno-osseous dysplasia patients have a small head circumference, and 15% have social, language, motor, or cognitive abnormalities. Postmortem examination of 2 Schimke immuno-osseous dysplasia patients showed low brain weights and subtle brain histologic abnormalities suggestive of perturbed neuron-glial migration such as heterotopia, irregular cortical thickness, incomplete gyral formation, and poor definition of cortical layers. We found that SMARCAL1 is highly expressed in the developing and adult mouse and human brain, including neural precursors and neuronal lineage cells. These observations suggest that SMARCAL1 deficiency may influence brain development and function in addition to its previously recognized effect on cerebral circulation. © 2008 American Association of Neuropathologists, Inc.

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Neural stem cells: involvement in adult neurogenesis and CNS repair

岡野栄之

PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES 363 （ 1500 ） 2111 - 2122 2008年06月

研究論文（学術雑誌）, ISSN 0962-8436

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Introduction. Japan: its tradition and hot topics in biological sciences

岡野栄之

PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES 363 （ 1500 ） 2067 - 2069 2008年06月

研究論文（学術雑誌）, ISSN 0962-8436

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Neurologic phenotype of Schimke immuno-osseous dysplasia and neurodevelopmental expression of SMARCAL1

岡野栄之

JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY 67 （ 6 ） 565 - 577 2008年06月

研究論文（学術雑誌）, ISSN 0022-3069

Notch mediates the segmental specification of angioblasts in somites and their directed migration toward the dorsal aorta in avian embryos

岡野栄之

DEVELOPMENTAL CELL 14 （ 6 ） 890 - 901 2008年06月

研究論文（学術雑誌）, ISSN 1534-5807

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Musashil modulates mammary progenitor cell expansion through proliferin-mediated activation of the Wnt and Notch pathways

岡野栄之

MOLECULAR AND CELLULAR BIOLOGY 28 （ 11 ） 3589 - 3599 2008年06月

研究論文（学術雑誌）, ISSN 0270-7306

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Effects of static magnetic fields in biology: role of free radicals

岡野栄之

FRONTIERS IN BIOSCIENCE 13 6106 - 6125 2008年05月

研究論文（学術雑誌）, ISSN 1093-9946

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Neural RNA-binding protein Musashi1 inhibits translation initiation by competing with eIF4G for PABP

岡野栄之

JOURNAL OF CELL BIOLOGY 181 （ 4 ） 639 - 653 2008年05月

研究論文（学術雑誌）, ISSN 0021-9525

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Hzf regulates adipogenesis through translational control of C/EBP alpha

岡野栄之

EMBO JOURNAL 27 （ 10 ） 1481 - 1490 2008年05月

研究論文（学術雑誌）, ISSN 0261-4189

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Role of IL-6 in regulation of inflammation and stem cell differentiation in CNS trauma

Mukaino Masahiko, Nakamura Masaya, Okada Seiji, Toyama Yoshiaki, Liu Meigen, Okano Hideyuki

Nihon Rinshō Men'eki Gakkai kaishi = Japanese journal of clinical immunology 31 （ 2 ） 93 - 98 2008年04月

ISSN 0911-4300

Interleukin-6 (IL-6) has been demonstrated to play a pivotal role in CNS trauma as a proinflammatory cytokine which regulates inflammatory response. Not only being a mediator of inflammation, but IL-6 induces neural stem/progenitor cells to undergo astrocytic differentiation selectively in the injured CNS. These effects are considered to coordinate to prevent the CNS repair after traumatic injury. Consistently, we previously reported that the administration of anti-IL-6 receptor antibody (MR16-1) immediately after spinal cord injury in mice decreased the number of invading inflammatory cells and the severity of connective tissue scar formation, and led to improved functional recovery. These findings suggest that neutralization of IL-6 signaling in the acute phase of SCI will be beneficial for CNS repair. A critical merit of this anti-IL-6 receptor antibody is that humanized antibody to human IL-6 receptor (MRA; Atlizumab) has already been reshaped. However, there are several studies which show beneficial aspects of IL-6 signaling in the pathology of CNS trauma. Further investigation of the mechanisms how MR16-1 reduces tissue damage should be required for clinical application.

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Isolation and characterization of dendritic cells from common marmosets for preclinical cell therapy studies

岡野栄之

IMMUNOLOGY 123 （ 4 ） 566 - 574 2008年04月

研究論文（学術雑誌）, ISSN 0019-2805

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Spatial gradient effects of 120 mT static magnetic field on endothelial tubular formation in vitro

岡野栄之

BIOELECTROMAGNETICS 29 （ 3 ） 233 - 236 2008年04月

研究論文（学術雑誌）, ISSN 0197-8462

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M-numb and notch signals are the key factors for gastric mucosal regeneration after acute damage

岡野栄之

GASTROENTEROLOGY 134 （ 4 ） A235 - A235 2008年04月

研究論文（学術雑誌）, ISSN 0016-5085

Cell-cycle-specific nestin expression coordinates with morphological changes in embryonic cortical neural progenitors

岡野栄之

JOURNAL OF CELL SCIENCE 121 （ 8 ） 1204 - 1212 2008年04月

研究論文（学術雑誌）, ISSN 0021-9533

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Ontogeny and multipotency of neural crest-derived stem cells in mouse bone marrow, dorsal root ganglia, and whisker pad

岡野栄之

CELL STEM CELL 2 （ 4 ） 392 - 403 2008年04月

研究論文（学術雑誌）, ISSN 1934-5909

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[Mammalian neural stem cells].

Shimazaki Takuya, Okano Hideyuki

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 53 （ 4 Suppl ） 311 - 317 2008年03月

ISSN 0039-9450

Enhanced transfection efficiency in laser-induced stress wave-assisted gene transfer at low laser fluence by increasing pressure impulse

岡野栄之

APPLIED PHYSICS EXPRESS 1 （ 3 ） S89 - S89 2008年03月

研究論文（学術雑誌）, ISSN 1882-0778

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[Strategies for axonal regeneration after spinal cord injury].

Kitamura Kazuya, Nakamura Masaya, Toyama Yoshiaki, Okano Hideyuki

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 53 （ 4 Suppl ） 411 - 417 2008年03月

ISSN 0039-9450

Helicobacter pylori infection induces candidate stem cell marker musashi-1 in the human gastric epithelium

岡野栄之

DIGESTIVE DISEASES AND SCIENCES 53 （ 2 ） 363 - 369 2008年02月

研究論文（学術雑誌）, ISSN 0163-2116

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The intestinal epithelium compensates for p53-mediated cell death and guarantees organismal survival

Valentin-Vega Y. A., Okano H., Lozano G.

Cell Death and Differentiation 15 （ 11 ） 1772 - 1781 2008年

ISSN 1350-9047

Mdm2 is the major inhibitor of the p53 tumor suppressor. Loss of Mdm2 in mice or in specific tissues of the mouse always yields p53-dependent lethal phenotypes. However, the role of Mdm2 in tissues with high turnover capacity is unknown. We have engineered mice lacking Mdm2 in the intestinal epithelium using the Cre/LoxP system. Loss of Mdm2 (Mdm2^intΔ) results in viable animals, but neonates display multiple intestinal abnormalities such as hyperplasia, enterocyte vacuolization, and inflammation. These defects correlate with a drastic increase in p53-dependent apoptosis in highly proliferative and differentiated cells. Unexpectedly, the observed phenotypes disappear with age. The tissue selects against Mdm2-null cells and increases its proliferative capacity. Additionally, the intestinal stem and progenitor cell populations are enriched leading to an increase in crypt fission events. Enhanced proliferation is achieved by activation of the canonical Wnt and EGFR-mediated Ras/MAPK pathways. While Mdm2 is a critical inhibitor of p53 in the intestinal epithelium, the tissue employs a series of processes that compensate for cell death.

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The long-term live monitoring of transplanted human fetal neural stem/progenitor cells in vivo

岡野栄之

NEUROSCIENCE RESEARCH 61 S85 - S85 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Post-translational regulation of Musashi1 RNA-binding activity and its translational control

岡野栄之

NEUROSCIENCE RESEARCH 61 S154 - S154 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Isolation and characterization of neural crest-derived stem cells in mouse bone marrow

岡野栄之

NEUROSCIENCE RESEARCH 61 S36 - S36 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Expression and function of Galectin-1 in adult neurogenesis of the hippocampus

岡野栄之

NEUROSCIENCE RESEARCH 61 S239 - S239 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Visualization of neurogenesis with fluorescent proteins

岡野栄之

NEUROSCIENCE RESEARCH 61 S229 - S229 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Common marmoset (Callithrix jacchus) as a model system to study neuroscience

岡野栄之

NEUROSCIENCE RESEARCH 61 S14 - S14 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Effect of low oxygen environment on cultured human fetal neural stem/progenitor cells

岡野栄之

NEUROSCIENCE RESEARCH 61 S157 - S157 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Prospective isolation and functional elucidation of murine mesenchymal stem cells

岡野栄之

NEUROSCIENCE RESEARCH 61 S154 - S154 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Spinal cord injury: Emerging beneficial role of reactive astrocytes' migration

岡野栄之

INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY 40 （ 9 ） 1649 - 1653 2008年

研究論文（学術雑誌）, ISSN 1357-2725

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Multipotent neural crest-derived stem cells in the adult mouse cornea

岡野栄之

NEUROSCIENCE RESEARCH 61 S281 - S281 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Isolation and identification of mesenchymal stem cells in human bone marrow

岡野栄之

NEUROSCIENCE RESEARCH 61 S84 - S84 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Mechanism of axonal regeneration and degeneration in spinal cord injury and other CNS diseases

岡野栄之

NEUROSCIENCE RESEARCH 61 S8 - S8 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Analysis of inner ear-selective cis-regulatory enhancer of Sox21 gene

岡野栄之

NEUROSCIENCE RESEARCH 61 S103 - S103 2008年

研究論文（学術雑誌）, ISSN 0168-0102

The morphological maturation course of the cerebellar Purkinje cell generated from the mouse embryonic stem cell

岡野栄之

NEUROSCIENCE RESEARCH 61 S158 - S158 2008年

研究論文（学術雑誌）, ISSN 0168-0102

A practical device for pinpoint delivery of molecules into multiple neurons in culture

岡野栄之

NEUROSCIENCE RESEARCH 61 S211 - S211 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Directed neural differentiation of induced pluripotent stem cells

岡野栄之

NEUROSCIENCE RESEARCH 61 S44 - S44 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Switching mechanism to neural differentiation controlled by Musashi and Hu proteins.

岡野栄之

NEUROSCIENCE RESEARCH 61 S155 - S155 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Effects of granulocyte macrophage colony stimulating factor following spinal cord injury in mice

岡野栄之

NEUROSCIENCE RESEARCH 61 S237 - S237 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Hepatocyte growth factor (HGF) promotes endogenous repair and functional recovery after spinal cord injury: Preclinical trial from rodent to primate

岡野栄之

NEUROSCIENCE RESEARCH 61 S16 - S16 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Self-assembling peptide scaffold promotes the axonal growth in the injured spinal cord

岡野栄之

NEUROSCIENCE RESEARCH 61 S93 - S93 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Suppression of Oct4 by germ cell nuclear factor restricts pluripotency and promotes neural stem cell development in the early neural lineage

岡野栄之

NEUROSCIENCE RESEARCH 61 S231 - S231 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Analysis of a transcription factor involved in neural stem cell maintenance

岡野栄之

NEUROSCIENCE RESEARCH 61 S230 - S230 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Sox21 is required for adult neurogenesis in mouse hippocampus

岡野栄之

NEUROSCIENCE RESEARCH 61 S96 - S96 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Whole genome transcriptome profile of human embryonic stem cells and differentiated cells

岡野栄之

NEUROSCIENCE RESEARCH 61 S43 - S43 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Coordination between the nestin gene expression and the morphological alteration of cortical neural progenitor cells

岡野栄之

NEUROSCIENCE RESEARCH 61 S87 - S87 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Prospective isolation of living neural stem cells from mouse brain through surface N-glycan

岡野栄之

NEUROSCIENCE RESEARCH 61 S139 - S139 2008年

研究論文（学術雑誌）, ISSN 0168-0102

In vivo imaging of contusive spinal cord injury using diffusion tensor tractography in Non-human Primates

岡野栄之

NEUROSCIENCE RESEARCH 61 S64 - S64 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Kspot, a novel ubiquitin ligase, regulates synaptic activity

岡野栄之

NEUROSCIENCE RESEARCH 61 S218 - S218 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Altered participation in neural differentiation by HuD splice variants

岡野栄之

NEUROSCIENCE RESEARCH 61 S229 - S229 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Comparison between ES cell-derived neurogenic and gliogenic neurospheres as a source of transplantation for spinal cord injury

岡野栄之

NEUROSCIENCE RESEARCH 61 S43 - S43 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Young neurons migrate along the blood vessel scaffold in the regenerating postnatal brain

岡野栄之

NEUROSCIENCE RESEARCH 61 S58 - S58 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Spatiotemporal Recapitulation of Central Nervous System Development by Murine Embryonic Stem Cell-Derived Neural Stem/Progenitor Cells

岡野栄之

STEM CELLS 26 （ 12 ） 3086 - 3098 2008年

研究論文（学術雑誌）, ISSN 1066-5099

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A factor secreted from bone-marrow-derived endothelial progenitor cells promotes self-renewal of neural stem cells in mice

岡野栄之

NEUROSCIENCE RESEARCH 61 S86 - S86 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Application of q-space MRI to spinal cord in common marmoset

岡野栄之

NEUROSCIENCE RESEARCH 61 S277 - S277 2008年

研究論文（学術雑誌）, ISSN 0168-0102

A novel gamma-secretase inhibitor, LY411575, replaced auditory hair cells and recovered hearing loss after severe acoustic trauma in mice

岡野栄之

NEUROSCIENCE RESEARCH 61 S25 - S25 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Proliferation of neural progenitors and induction of reactive glia after brain ischemia detected with Nestin-Cre-ER system

岡野栄之

NEUROSCIENCE RESEARCH 61 S133 - S133 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Neural crest-derived cells in the peripheral olfactory system

岡野栄之

NEUROSCIENCE RESEARCH 61 S178 - S178 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Angiotensin II type 1 receptor signaling contributes to synaptophysin degradation and neuronal dysfunction in the diabetic retina

岡野栄之

NEUROSCIENCE RESEARCH 61 S220 - S220 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Analysis of transcriptional regulation of musashi1 gene in neural stem cells

岡野栄之

NEUROSCIENCE RESEARCH 61 S152 - S152 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Comparative study of different administrations of the neural stem cells for spinal cord injury in mice

岡野栄之

NEUROSCIENCE RESEARCH 61 S85 - S85 2008年

研究論文（学術雑誌）, ISSN 0168-0102

In vitro and vivo characterization in human neural stem/progenitor cells expressing ABC transporter ABCB1

岡野栄之

NEUROSCIENCE RESEARCH 61 S86 - S86 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Functional analysis of Hu-associated RNA binding protein-network

岡野栄之

NEUROSCIENCE RESEARCH 61 S42 - S42 2008年

研究論文（学術雑誌）, ISSN 0168-0102

The effect of anti-interleukin-6 receptor antibody in noise induced damaged cochlea

岡野栄之

NEUROSCIENCE RESEARCH 61 S102 - S102 2008年

研究論文（学術雑誌）, ISSN 0168-0102

The functional analysis of RNA-binding protein Musashi2 in the cerebellum

岡野栄之

NEUROSCIENCE RESEARCH 61 S89 - S89 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Investigation of neural crest stem cell therapy for Hirschsprung's disease

岡野栄之

NEUROSCIENCE RESEARCH 61 S36 - S36 2008年

研究論文（学術雑誌）, ISSN 0168-0102

SOCS3 is required to protect photoreceptor cell function during retinal inflammation

岡野栄之

NEUROSCIENCE RESEARCH 61 S269 - S269 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Neural RNA-binding protein Musashi1 inhibits translation initiation by competing with eIF4G for PABP

岡野栄之

NEUROSCIENCE RESEARCH 61 S153 - S153 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Anti IL-6 antibody treatment reduces the infiltration of hematogenous macrophages into the injured spinal cord

岡野栄之

NEUROSCIENCE RESEARCH 61 S275 - S275 2008年

研究論文（学術雑誌）, ISSN 0168-0102

A role of neuronal RNA-binding protein Hu and NF45/90 in the nervous system

岡野栄之

NEUROSCIENCE RESEARCH 61 S153 - S153 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Expression of free fatty acid receptor GPR40 in the neurogenic niche of adult monkey hippocampus

岡野栄之

HIPPOCAMPUS 18 （ 3 ） 326 - 333 2008年

研究論文（学術雑誌）, ISSN 1050-9631

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Mature Adipocyte-Derived Cells, Dedifferentiated Fat Cells (DFAT), Promoted Functional Recovery From Spinal Cord Injury-Induced Motor Dysfunction in Rats

岡野栄之

CELL TRANSPLANTATION 17 （ 8 ） 877 - 886 2008年

研究論文（学術雑誌）, ISSN 0963-6897

In vivo imaging in humanized mice

Masuda H., Okano H. J., Maruyama T., Yoshimura Y., Okano H., Matsuzaki Y.

Current topics in microbiology and immunology 324 179 - 196 2008年

ISSN 0070-217X

The radiological modalities that are currently utilized as critical components in clinical medicine have also been adapted to small-animal imaging, among which are ultrasound imaging, X-ray computerized tomography (CT), magnetic resonance imaging (MRI), positron emission tomography (PET), and singlephoton emission computed tomography (SPECT). Optical imaging techniques such as bioluminescence imaging (BLI) and fluorescence imaging (FLI) are approaches that are commonly used in small animals. Longitudinal surveys of living (i.e., nonsacrificed) animal models with these modalities provide some clues for the development of clinical applications. The techniques are absolutely essential for translational research. However, there are currently few tools available with sufficient spatial or temporal resolution ideal for all experimental studies. In this chapter, we provide a rationale and techniques for visualizing target cells in living small animals and an overview of the advantages and limitations of current imaging technology. Finally, we introduce a humanized mouse and a novel in vivo imaging system that we have developed. We also discuss real-time observations of reconstructs and clinical manifestations. © 2008 Springer-Verlag.

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Structural analysis of Musashi-RNA complex on the basis of long-range structural information.

Ohyama Takako, Furukawa Ayako, Mashima Tsukasa, Sugiyama Takashi, Ohgara Shouta, Yamazaki Toshio, Imai Takao, Okano Hideyuki, Nagata Takashi, Katahira Masato

Nucleic acids symposium series (2004) （ 52 ） 193 - 194 2008年

Musashi protein is supposed to be involved in the regulation of differentiation of neural stem cells. Musashi binds to 3' untranslated region of target mRNA and represses the translation of mRNA. Musashi has two tandem RNA-binding domains (RBDs), RBD1 and RBD2. Both RBDs cooperatively bind to the target mRNA. Here, we determined the structure of RBD1-RBD2 in complex with target RNA. First, the structures of two RBDs in the complex were determined on the basis of short-range distance restrains derived from NOEs. However, the relative position of two RBDs was not determined due to the lack of long-range distance restraints across two RBDs. In order to overcome the situation, we introduced the paramagnetic center into Musashi by attaching MTSL carrying the NO radical. The long-range distance restraints (ca. 20-40 A) between two RBDs were derived from paramagnetic relaxation enhancement (PRE) caused by the paramagnetic center. The relative position of two RBDs was successfully determined on the basis of these long-range distance restraints. The change in the relative position of two RBDs on binding to the target RNA was also detected by PRE. The determined structure of RBD1-RBD2 in the complex has suggested how Musashi recognizes its target mRNA.

Mature adipocyte-derived cells, dedifferentiated fat cells (DFAT), promoted functional recovery from spinal cord injury-induced motor dysfunction in rats

Ohta Yuki, Takenaga Mitsuko, Tokura Yukie, Hamaguchi Akemi, Matsuraoto Taro, Kano Koichiro, Mugishima Hideo, Okano Hideyuki, Igarashi Rie

Cell Transplantation 17 （ 8 ） 877 - 886 2008年

ISSN 0963-6897

Transplantation of mature adipocyte-derived cells (dedifferentiated fat cells) led to marked functional recovery from spinal cord injury (SCI)-induced motor dysfunction in rats. When mature adipocytes were isolated from rat adipose tissue and grown in ceiling culture, transformation into fibroblast-like cells without lipid droplets occurred. These fibroblast-like cells, termed dedifferentiated fat cells (DFAT), could proliferate and could also differentiate back into adipocytes. DFAT expressed neural markers such as nestin, βIII tubulin, and GFAP. Allografting of DFAT into SCI-induced rats led to significant recovery from hindlimb dysfunction. Grafted cells were detected at the injection site, and some of these cells expressed βIII tubulin. DFAT expressed neurotrophic factors such as BDNF and GDNF prior to transplantation, and grafted cells were also positive for these factors. Therefore, these neurotrophic factors derived from grafted DFAT might have contributed to the promotion of functional recovery. These findings also suggest that mature adipocytes could become a new source for cell replacement therapy to treat central nervous system disorders. Copyright © 2008 Cognizant Comm. Corp.

Cell therapy for spinal cord injury by neural stem/progenitor cells derived from induced pluripotent stem cells

岡野栄之

NEUROSCIENCE RESEARCH 61 S69 - S69 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Neural RNA-binding protein Musashi1 inhibits translation initiation by competing with eIF4G for PABP

岡野栄之

NEUROSCIENCE RESEARCH 61 S153 - S153 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Expression of free fatty acid receptor GPR40 in the neurogenic niche of adult monkey hippocampus

岡野栄之

HIPPOCAMPUS 18 （ 3 ） 326 - 333 2008年

研究論文（学術雑誌）, ISSN 1050-9631

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Analysis of RNA binding protein Musashi2 in mammalian nervous system development

岡野栄之

NEUROSCIENCE RESEARCH 61 S89 - S89 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Stromal cell-secreted factors promote the survival of ES cell-derived early neural stem/progenitor cells via the activation of MAPK and PI3K-Akt pathways

岡野栄之

NEUROSCIENCE RESEARCH 61 S156 - S156 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Neuroepithelial stem cells are converted into neural progenitors during an elongated interphase in Drosophila larval optic lobe

岡野栄之

NEUROSCIENCE RESEARCH 61 S156 - S156 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Mechanisms of reactive astrocytes migration after spina cord injury

岡野栄之

NEUROSCIENCE RESEARCH 61 S227 - S227 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Intraarterial transplantation of mononuclear cells promotes neovascularization after transient cerebral ischemia in mice

岡野栄之

NEUROSCIENCE RESEARCH 61 S134 - S134 2008年

研究論文（学術雑誌）, ISSN 0168-0102

An efficient screening method using inner-ear-derived spheres for selection of compounds that induce hair cell differentiation

岡野栄之

NEUROSCIENCE RESEARCH 61 S57 - S57 2008年

研究論文（学術雑誌）, ISSN 0168-0102

Notch-Hes1 Pathway Contributes to the Cochlear Prosensory Formation Potentially Through the Transcriptional Down-Regulation of p27(Kip1)

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 87 （ 16 ） 3521 - 3534 2009年12月

研究論文（学術雑誌）, ISSN 0360-4012

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Preimplantation Development of Somatic Cell Cloned Embryos in the Common Marmoset (Callithrix jacchus)

岡野栄之

CLONING AND STEM CELLS 11 （ 4 ） 575 - 583 2009年12月

研究論文（学術雑誌）, ISSN 1536-2302

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Transplantation of dendritic cells promotes functional recovery from spinal cord injury in common marmoset

岡野栄之

NEUROSCIENCE RESEARCH 65 （ 4 ） 384 - 392 2009年12月

研究論文（学術雑誌）, ISSN 0168-0102

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Neuronal re-juvenilization in the nucleus ambiguus after vagal nerve injury

岡野栄之

NEUROSCIENCE RESEARCH 65 （ 4 ） 353 - 359 2009年12月

研究論文（学術雑誌）, ISSN 0168-0102

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Reconsideration of Insulin Signals Induced by Improved Laboratory Animal Diets, Japanese and American Diets, in IRS-2 Deficient Mice

Hashimoto H., Arai T., Mori A., Kawai K., Hikishima K., Ohnishi Y., Eto T., Ito M., Hioki K., Suzuki R., Ohsugi M., Saito M., Ueyama Y., Okano H., Yamauchi T., Kubota N., Ueki K., Tobe K., Tamaoki N., Kadowaki T., Kosaka K.

Experimental and Clinical Endocrinology and Diabetes 117 （ 10 ） 577 - 586 2009年11月

ISSN 0947-7349

Current Japanese and American diets and Japanese diet immediately after the War were converted to laboratory animal diets. As a result, current laboratory animal diet (CA-I, CLEA) unexpectedly resembled the diet of Japanese after the War. This is considered to result in an under-evaluation of diabetes research using laboratory animals at present. Therefore, changes in insulin signals caused by current Japanese and American diets were examined using IRS-2 deficient mice (Irs2 ^-/- mice) and mechanisms of aggravation of type 2 diabetes due to modern diets were examined. Irs2 ^-/- mice at 6 weeks of age were divided into three groups: Japanese diet (Jd) group, American diet (Ad) group and CA-I diet [regular diet (Rd)] group. Each diet was given to the dams from 7 days before delivery. When the Irs2 ^-/- mice reached 6 weeks of age, the glucose tolerance test (GTT), insulin tolerance test (ITT) and organ sampling were performed. The sampled organs and white adipose tissue were used for analysis of RNA, enzyme activity and tissues. In GTT and ITT, the Ad group showed worse glucose tolerance and insulin resistance than the Rd group. Impaired glucose tolerance of the Jd group was the same as that of the Rd group, but insulin resistance was worse than in the Rd group. These results were caused an increase in fat accumulation and adipocytes in the peritoneal cavity by lipogenic enzyme activity in the liver and muscle, and the increase in TNFa of hypertrophic adipocyte origin further aggravated insulin resistance and the increase in resistin also aggravated the impaired glucose tolerance, leading to aggravation of type 2 diabetes. The Japanese and American diets given to the Irs2 ^-/- mice, which we developed, showed abnormal findings in some Irs2 ^-/- mice but inhibited excessive reactions of insulin signals as diets used in ordinary nutritional management.

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Roles of ES Cell-Derived Gliogenic Neural Stem/Progenitor Cells in Functional Recovery after Spinal Cord Injury

岡野栄之

PLOS ONE 4 （ 11 ） 1109 - 1119 2009年11月

研究論文（学術雑誌）, ISSN 1932-6203

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Strategies toward CNS-regeneration using induced pluripotent stem cells.

Okano Hideyuki

Genome informatics. International Conference on Genome Informatics 23 （ 1 ） 217 - 220 2009年10月

ISSN 0919-9454

Induced pluripotent stem (iPS) cells are pluripotent stem cells directly reprogrammed from cultured mouse fibroblast by introducing Oct3/4, Sox2, c-Myc, and Klf4. Cells obtained using this technology, which allows the ethical issues and immunological rejection associated with embryonic stem (ES) cells to be avoided, might be a clinically useful source for cell replacement therapics. Here we demonstrate that murine iPS cells formed neurospheres that produced electrophysiologically functional neurons, astrocytes, and oligodendrocytes. Secondary neurospheres (SNSs) generated from various mouse iPS cell showed their neural differentiation capacity and teratoma formation after transplantation into the brain of immunodeficient NOD/SCID mice. We found that origin (source of somatic cells) of the iPS cells are the crucial determinant for the potential tumorigenicity of iPS-derived neural stem/progenitor cclls and that their tumorigenicity results from the persistent presence of undifferentiated cells within the SNSs. Furthermore, transplantation of non-tumorigenic Nanog-iPS-derived SNSs into mouse spinal cord injury (SCI) model promoted locomotor function recovery. Surprisingly, SNSs derived from c-Myc minus iPS cells generated without drug selection showed robust tumorigenesis, in spite of their potential to contribute adult chimeric mice without tumor formation.

Prospective identification, isolation, and systemic transplantation of multipotent mesenchymal stem cells in murine bone marrow

岡野栄之

JOURNAL OF EXPERIMENTAL MEDICINE 206 （ 11 ） 2483 - 2496 2009年10月

研究論文（学術雑誌）, ISSN 0022-1007

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Identification of tumor-initiating cells in a highly aggressive brain tumor using promoter activity of nucleostemin

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 106 （ 40 ） 17163 - 17168 2009年10月

研究論文（学術雑誌）, ISSN 0027-8424

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ENVIRONMENTAL CUE-DEPENDENT DOPAMINERGIC NEURONAL DIFFERENTIATION AND FUNCTIONAL EFFECT OF GRAFTED NEUROEPITHELIAL STEM CELLS IN PARKINSONIAN BRAIN

岡野栄之

NEUROSURGERY 65 （ 4 ） 741 - 753 2009年10月

研究論文（学術雑誌）, ISSN 0148-396X

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Cell surface N-glycans mediated isolation of mouse neural stem cells

岡野栄之

JOURNAL OF NEUROCHEMISTRY 110 （ 5 ） 1575 - 1584 2009年09月

研究論文（学術雑誌）, ISSN 0022-3042

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ABCB1 Is Predominantly Expressed in Human Fetal Neural Stem/progenitor Cells at an Early Development Stage

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 87 （ 12 ） 2615 - 2623 2009年09月

研究論文（学術雑誌）, ISSN 0360-4012

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Selective induction of neocortical GABAergic neurons by the PDK1-Akt pathway through activation of Mash1

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 106 （ 31 ） 13064 - 13069 2009年08月

研究論文（学術雑誌）, ISSN 0027-8424

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RMD-1, a novel microtubule-associated protein, functions in chromosome segregation in Caenorhabditis elegans (vol 179, pg 1149, 2007)

岡野栄之

JOURNAL OF CELL BIOLOGY 186 （ 4 ） 629 - 629 2009年08月

研究論文（学術雑誌）, ISSN 0021-9525

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Neural Crest-Derived Stem Cells Display a Wide Variety of Characteristics

岡野栄之

JOURNAL OF CELLULAR BIOCHEMISTRY 107 （ 6 ） 1046 - 1052 2009年08月

研究論文（学術雑誌）, ISSN 0730-2312

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Variation in the safety of induced pluripotent stem cell lines

岡野栄之

NATURE BIOTECHNOLOGY 27 （ 8 ） 743 - 745 2009年08月

研究論文（学術雑誌）, ISSN 1087-0156

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Fbxo45 Forms a Novel Ubiquitin Ligase Complex and Is Required for Neuronal Development

岡野栄之

MOLECULAR AND CELLULAR BIOLOGY 29 （ 13 ） 3529 - 3543 2009年07月

研究論文（学術雑誌）, ISSN 0270-7306

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3 '-Untranslated region of doublecortin mRNA is a binding target of the Musashi1 RNA-binding protein

岡野栄之

FEBS LETTERS 583 （ 14 ） 2429 - 2434 2009年07月

研究論文（学術雑誌）, ISSN 0014-5793

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Possibility of iPS cells (discussion)

Okano Hideyuki, Yamanaka Shinya, Tsuji Shoji

Brain and nerve = Shinkei kenkyū no shinpo 61 （ 6 ） 711 - 718 2009年06月

ISSN 1881-6096

Drosophila big brain does not act as a water channel, but mediates cell adhesion

岡野栄之

FEBS LETTERS 583 （ 12 ） 2077 - 2082 2009年06月

研究論文（学術雑誌）, ISSN 0014-5793

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The disruption of Sox21-mediated hair shaft cuticle differentiation causes cyclic alopecia in mice

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 106 （ 23 ） 9292 - 9297 2009年06月

研究論文（学術雑誌）, ISSN 0027-8424

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Generation of transgenic non-human primates with germline transmission

岡野栄之

NATURE 459 （ 7246 ） 523 - U50 2009年05月

研究論文（学術雑誌）, ISSN 0028-0836

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Tumor initiating potential of side population cells in human gastric cancer

岡野栄之

INTERNATIONAL JOURNAL OF ONCOLOGY 34 （ 5 ） 1201 - 1207 2009年05月

研究論文（学術雑誌）, ISSN 1019-6439

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M-Numb Expression Induced Gastric Mucosal Regeneration After Injury By Stimulating the Expression of Morphogenic Genes

岡野栄之

GASTROENTEROLOGY 136 （ 5 ） A23 - A23 2009年05月

研究論文（学術雑誌）, ISSN 0016-5085

Effect of a Gradient Static Magnetic Field on an Unstirred Belousov-Zhabotinsky Reaction by Changing the Thickness of the Medium

岡野栄之

JOURNAL OF PHYSICAL CHEMISTRY A 113 （ 13 ） 3061 - 3067 2009年04月

研究論文（学術雑誌）, ISSN 1089-5639

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Cell types to order: temporal specification of CNS stem cells

岡野栄之

CURRENT OPINION IN NEUROBIOLOGY 19 （ 2 ） 112 - 119 2009年04月

研究論文（学術雑誌）, ISSN 0959-4388

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Musashi-1, an RNA-binding protein, is indispensable for survival of photoreceptors

岡野栄之

EXPERIMENTAL EYE RESEARCH 88 （ 3 ） 347 - 355 2009年03月

研究論文（学術雑誌）, ISSN 0014-4835

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The Role of Uterine Smooth Muscle Stem Cells, Isolated Based on Cell Surface Markers, in Myometrial Remodeling in the Pregnant Uterus

岡野栄之

REPRODUCTIVE SCIENCES 16 （ 3 ） 296A - 297A 2009年03月

研究論文（学術雑誌）, ISSN 1933-7191

Adipogenic differentiation of human induced pluripotent stem cells: Comparison with that of human embryonic stem cells

岡野栄之

FEBS LETTERS 583 （ 6 ） 1029 - 1033 2009年03月

研究論文（学術雑誌）, ISSN 0014-5793

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The effect of Am-80, a synthetic retinoid, on spinal cord injury-induced motor dysfunction in rats

Takenaga Mitsuko, Ohta Yuki, Tokura Yukie, Hamaguchi Akemi, Shudo Koichi, Okano Hideyuki, Igarashi Rie

Biological and Pharmaceutical Bulletin 32 （ 2 ） 225 - 231 2009年02月

ISSN 0918-6158

The present study investigated the effect of 4[(5,6,7,8-tetrahydro-5,5,8,8, -tetramethyl-2-naphthalenyl)car-bamoyl] benzoic acid (Am-80), a synthetic retinoid, on spinal cord injury (SCI) in rats. Treatment with Am-80 (orally and subcutaneously) significantly promoted recovery from SCI-induced motor dysfunction. On day 28 after injury, the lesion cavity was markedly reduced, while the expression of myelin basic protein (MBP; myelin), βIIItubulin (neuron), and glial fibrillary acidic protein (GFAP; astrocyte) was increased, in comparison with SCI controls. Interestingly, expression of neurotrophin receptor, tyrosine kinase B (TrkB) was over 3-fold higher after Am-80 treatment than in SCI controls. A lot of TrkB-positive cells as well as brain-derived neurotrophic factor (BDNF)-positive ones were observed around the injured site. Am-80 (10 μm) combined with BDNF (100ng/ml) promoted extensive neurite outgrowth and TrkB gene expression by cultured SH-SY5Y cells, as did all-trans retinoic acid (ATRA). Thymidine incorporation was dramatically suppressed, but there was little effect on cell viability. These findings suggest that Am-80 has the potential to be used for treating neurodegenerative disorders, including SCI. Its efficacy may be partly ascribed to promotion of cell viability and differentiation of neural stem cells through increased TrkB expression. © 2009 Pharmaceutical Society of Japan.

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Visualization of peripheral nerve degeneration and regeneration: Monitoring with diffusion tensor tractography

岡野栄之

NEUROIMAGE 44 （ 3 ） 884 - 892 2009年02月

研究論文（学術雑誌）, ISSN 1053-8119

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Development of mesenchymal stem cells partially originate from the neural crest

岡野栄之

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 379 （ 4 ） 1114 - 1119 2009年02月

研究論文（学術雑誌）, ISSN 0006-291X

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The Effect of Am-80, a Synthetic Retinoid, on Spinal Cord Injury-Induced Motor Dysfunction in Rats

岡野栄之

BIOLOGICAL & PHARMACEUTICAL BULLETIN 32 （ 2 ） 225 - 231 2009年02月

研究論文（学術雑誌）, ISSN 0918-6158

Suppression of Oct4 by Germ Cell Nuclear Factor Restricts Pluripotency and Promotes Neural Stem Cell Development in the Early Neural Lineage

岡野栄之

JOURNAL OF NEUROSCIENCE 29 （ 7 ） 2113 - 2124 2009年02月

研究論文（学術雑誌）, ISSN 0270-6474

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Evaluation of Human Fetal Neural Stem/Progenitor Cells as a Source for Cell Replacement Therapy for Neurological Disorders: Properties and Tumorigenicity After Long-Term In Vitro Maintenance

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 87 （ 2 ） 307 - 317 2009年02月

研究論文（学術雑誌）, ISSN 0360-4012

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Mouse Embryonic Stem Cells Give Rise to Gut-Like Morphogenesis, Including Intestinal Stem Cells, in the Embryoid Body Model

岡野栄之

STEM CELLS AND DEVELOPMENT 18 （ 1 ） 113 - 125 2009年01月

研究論文（学術雑誌）, ISSN 1547-3287

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Regeneration of the central nervous system using iPS cell-technologies

Okano Hideyuki

Clinical Neurology 49 （ 11 ） 825 - 826 2009年

ISSN 0009-918X

Induced pluripotent stem (iPS) cells are pluripotent stem cells directly reprogrammed from cultured mouse fibroblast by introducing Oct3/4, Sox2, c-Myc, and Klf4. Cells obtained using this technology, which allows the ethical issues and immunological rejection associated with embryonic stem (ES) cells to be avoided, might be a clinically useful source for cell replacement therapies. We found that secondary neurospheres (SNSs) generated from various mouse iPS cell showed their neural differentiation capacity and teratoma formation after transplantation into the brain of immunodeficient NOD/SCID mice. We found that origin (source of somatic cells) of the iPS cells are the crucial determinant for the potential tumorigenicity of iPS-derived neural stem/progenitor cells and that their tumorigenicity results from the persistent presence of undifferentiated cells within the SNSs. Surprisingly, SNSs derived from c-Myc minusiPS cells generated without drug selection showed robust tumorigenesis, in spite of their potential to contribute adult chimeric mice without tumor formation. Furthermore, we examined whether the transplantation of non-tumorigenic Nanog-iPS-derived SNSs into mouse spinal cord injury (SCI) model could promote locomotor function recovery. As a result, we found that properly pre-evaluated iPS clone-derived neural stem/progenitor cells may be a promising cell source for future transplantation therapy of SCI.

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COMPARATIVE STUDY OF DIFFERENT ADMINISTRATIONS OF THE NEURAL STEM CELLS FOR SPINAL CORD INJURY IN MICE

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 442 - 442 2009年

研究論文（学術雑誌）, ISSN 1880-6546

SENSORY NEURAL NETWORK FORMATION WITH RNA BINDING PROTEIN MUSASHI2 DEFICIENT MICE

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 337 - 337 2009年

研究論文（学術雑誌）, ISSN 1880-6546

ANALYSIS OF CELL SURFACE MOLECULES WITH COMPLEX TYPE N-GLYCANS ON NEURAL STEM CELLS

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 291 - 291 2009年

研究論文（学術雑誌）, ISSN 1880-6546

Distinct roles of Sox family transcription factors in adult neurogenesis

岡野栄之

NEUROSCIENCE RESEARCH 65 S5 - S5 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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Macrophage migration inhibitory factor (MIF) promotes proliferation and cell survival of neural stem/progenitor cells

岡野栄之

NEUROSCIENCE RESEARCH 65 S91 - S91 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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STAMPORATION: A NEW MICROINJECTION METHOD INTO VERIOUS CULTURED CELLS

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 528 - 528 2009年

研究論文（学術雑誌）, ISSN 1880-6546

Planar polarity decisions for directional beating of ependymal cilia and fluid flow in the adult mouse lateral ventricles

岡野栄之

NEUROSCIENCE RESEARCH 65 S54 - S54 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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NEURAL DIFFERENTIATION AND CELL THERAPY USING PLURIPOTENT STEM CELLS

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 45 - 45 2009年

研究論文（学術雑誌）, ISSN 1880-6546

Possible implication of mu-opioid receptors in the neural differentiation from embryonic stem cells

岡野栄之

NEUROSCIENCE RESEARCH 65 S137 - S137 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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SLIT-ROBO SIGNALING REGULATES THE MIGRATION OF NEW NEURONS UNDER PHYSIOLOGICAL AND PATHOLOGICAL CONDITIONS

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 442 - 442 2009年

研究論文（学術雑誌）, ISSN 1880-6546

THE CHARACTERIZATION OF THE CEREBELLAR PURKINJE CELLS INDUCED FROM THE MICE EMBRYONIC STEM CELLS AND TRANSPLANTATION INTO THE ADULT PCD MICE

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 343 - 343 2009年

研究論文（学術雑誌）, ISSN 1880-6546

Sensory network formation of RNA binding protein Musashi2

岡野栄之

NEUROSCIENCE RESEARCH 65 S96 - S96 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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Role of Diversin as a possible switch of distinct Wnt pathways regulating multiple steps of adult neurogenesis

岡野栄之

NEUROSCIENCE RESEARCH 65 S54 - S54 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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ROLES OF RNA BINDING PROTEIN Hu IN DEVELOPING AND MATURE NEURONS

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 343 - 343 2009年

研究論文（学術雑誌）, ISSN 1880-6546

ROLE OF WNT/BETA-CATENIN SIGNALING IN NEUROGENESIS IN THE ADULT MOUSE SUBVENTRICULAR ZONE

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 442 - 442 2009年

研究論文（学術雑誌）, ISSN 1880-6546

DIFFUSION TENSOR PERIPHERAL NERVE TRACTOGRAPHY

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 293 - 293 2009年

研究論文（学術雑誌）, ISSN 1880-6546

Promotion of the neural differentiation from embryonic stem cells activated by dopamine D-3 receptors

岡野栄之

NEUROSCIENCE RESEARCH 65 S137 - S137 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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A role of neuronal RNA binding protein Hu and NF45/NF90 in the nervous system

岡野栄之

NEUROSCIENCE RESEARCH 65 S90 - S90 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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Genetic analysis of the organization and function of the blood brain barrier in Drosophila

岡野栄之

NEUROSCIENCE RESEARCH 65 S150 - S150 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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Strategies toward regenerating the damaged CNS using ES/iPS cells

岡野栄之

NEUROSCIENCE RESEARCH 65 S1 - S1 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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Neuroepithelial stem cells are converted into neural progenitors during an elongated interphase in drosophila larval optic lobe

岡野栄之

JOURNAL OF NEUROGENETICS 23 S19 - S19 2009年

研究論文（学術雑誌）, ISSN 0167-7063

Analysis of cell surface glycoproteins on neural stem cells

岡野栄之

NEUROSCIENCE RESEARCH 65 S90 - S91 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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AXONAL DEGENERATION IN HuC-DEFICIENT CEREBELLUM

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 337 - 337 2009年

研究論文（学術雑誌）, ISSN 1880-6546

ROLE AND MECHANISMS OF REACTIVE ASTROCYTE S MIGRATION AFTER SPINAL CORD INJURY

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 341 - 341 2009年

研究論文（学術雑誌）, ISSN 1880-6546

Analysis of spinal cord injury using diffusion MRI

岡野栄之

NEUROSCIENCE RESEARCH 65 S35 - S35 2009年

研究論文（学術雑誌）, ISSN 0168-0102

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COMPARATIVE STUDY OF DIFFERENT ADMINISTRATIONS OF THE NEURAL STEM CELLS FOR SPINAL CORD INJURY IN MICE

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 442 - 442 2009年

研究論文（学術雑誌）, ISSN 1880-6546

A ROLE FOR RNA-BINDING PROTEIN MUSASHI 1 IN REGULATING THE AXONAL MIDLINE CROSSING

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 342 - 342 2009年

研究論文（学術雑誌）, ISSN 1880-6546

Epigenetic mechanisms regulating neural cell fate determination

Nakashima Kinichi, Kohyama Jun, Namihira Masakazu, Gage Fred H., Okano Hideyuki, Sawamoto Kazunobu

No To Hattatsu 41 （ 6 ） 411 - 414 2009年

ISSN 0029-0831

Neural stem/progenitor cells (NSCs/NPCs) give rise to neurons, astrocytes, and oligodendrocytes. It has become apparent that intracellular epigenetic modification including DNA methylation, in concert with extracellular cues such as cytokine signaling, is deeply involved in specifiying the fate of NSCs/NPCs by defining cell-type specific gene expression. However, it is still unclear how differentiated neural cells retain their specific attributes by repressing cellular properties characteristic of other lineages. In previous work, we have shown that methyl-CpG binding protein transcriptional repressors (MBDs), which were expressed predominantly in neurons in the central nervous system, inhibited astrocyte-specific gene expression by binding to highly methylated regions of their target genes. Here we report that oligodendrocytes, which do not express MBDs, can transdifferentiate into astrocytes both in vitro (cytokine stimulation) and in vivo (ischemic injury) through the activation of the JAK/STAT signaling pathway. These findings suggest that differentiation plasticity in neural cells is regulated by cell-intrinsic epigenetic mechanisms in collaboration with ambient cell-extrinsic cues.

RETROVIRUS INTEGRATION SITE ANALYSIS OF HUMAN INDUCED PLURIPOTENT STEM (IPS) CELLS

岡野栄之

JOURNAL OF GENE MEDICINE 12 （ 12 ） 1060 - 1061 2010年12月

研究論文（学術雑誌）, ISSN 1099-498X

Identification of Genes Required for Neural-Specific Glycosylation Using Functional Genomics

岡野栄之

PLOS GENETICS 6 （ 12 ） S57 - S57 2010年12月

研究論文（学術雑誌）, ISSN 1553-7404

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[Frontline in the research of regenerative medicine].

Inoue Haruhisa, Okano Hideyuki

Rinshō shinkeigaku = Clinical neurology 50 （ 11 ） 886 2010年11月

Expression and Proliferation-Promoting Role of Diversin in the Neuronally Committed Precursor Cells Migrating in the Adult Mouse Brain

岡野栄之

STEM CELLS 28 （ 11 ） 2017 - 2026 2010年11月

研究論文（学術雑誌）, ISSN 1066-5099

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Balanced ubiquitylation and deubiquitylation of Frizzled regulate cellular responsiveness to Wg/Wnt

岡野栄之

DIFFERENTIATION 80 S61 - S61 2010年11月

研究論文（学術雑誌）, ISSN 0301-4681

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iPS based cell therapy for the damaged CNS

岡野栄之

DIFFERENTIATION 80 S13 - S13 2010年11月

研究論文（学術雑誌）, ISSN 0301-4681

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New insight into cancer therapeutics: Induction of differentiation by regulating the Musashi/Numb/Notch pathway

岡野栄之

CELL RESEARCH 20 （ 10 ） 1083 - 1085 2010年10月

研究論文（学術雑誌）, ISSN 1001-0602

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Benefits of Utilizing Gene-Modified iPSCs for Clinical Applications

岡野栄之

CELL STEM CELL 7 （ 4 ） 429 - 430 2010年10月

研究論文（学術雑誌）, ISSN 1934-5909

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Generating induced pluripotent stem cells from common marmoset (Callithrix jacchus) fetal liver cells using defined factors, including Lin28

岡野栄之

GENES TO CELLS 15 （ 9 ） 959 - 969 2010年09月

研究論文（学術雑誌）, ISSN 1356-9597

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Generating induced pluripotent stem cells from common marmoset (Callithrix jacchus) fetal liver cells using defined factors, including Lin28

岡野栄之

GENES TO CELLS 15 （ 9 ） 959 - 969 2010年09月

研究論文（学術雑誌）, ISSN 1356-9597

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FRS2 alpha Regulates Erk Levels to Control a Self-Renewal Target Hes1 and Proliferation of FGF-Responsive Neural Stem/Progenitor Cells

岡野栄之

STEM CELLS 28 （ 9 ） 1660 - 1672 2010年09月

研究論文（学術雑誌）, ISSN 1066-5099

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Planar polarity of multiciliated ependymal cells involves the anterior migration of basal bodies regulated by non-muscle myosin II

岡野栄之

DEVELOPMENT 137 （ 18 ） 3037 - 3046 2010年09月

研究論文（学術雑誌）, ISSN 0950-1991

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The Neural Stem/Progenitor Cell Marker Nestin Is Expressed in Proliferative Endothelial Cells, but Not in Mature Vasculature

岡野栄之

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY 58 （ 8 ） 721 - 730 2010年08月

研究論文（学術雑誌）, ISSN 0022-1554

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Musashi1 regulates breast tumor cell proliferation and is a prognostic indicator of poor survival

岡野栄之

MOLECULAR CANCER 9 2462 - 2469 2010年08月

研究論文（学術雑誌）, ISSN 1476-4598

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Neural RNA-Binding Protein Musashi1 Controls Midline Crossing of Precerebellar Neurons through Posttranscriptional Regulation of Robo3/Rig-1 Expression

岡野栄之

NEURON 67 （ 3 ） 407 - 421 2010年08月

研究論文（学術雑誌）, ISSN 0896-6273

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Anti-IL-6-receptor antibody promotes repair of spinal cord injury by inducing microglia-dominant inflammation

岡野栄之

EXPERIMENTAL NEUROLOGY 224 （ 2 ） 403 - 414 2010年08月

研究論文（学術雑誌）, ISSN 0014-4886

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ADF2 is required for transformation of the ookinete and sporozoite in malaria parasite development

岡野栄之

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 397 （ 4 ） 668 - 672 2010年07月

研究論文（学術雑誌）, ISSN 0006-291X

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New Neurons Clear the Path of Astrocytic Processes for Their Rapid Migration in the Adult Brain

岡野栄之

NEURON 67 （ 2 ） 213 - 223 2010年07月

研究論文（学術雑誌）, ISSN 0896-6273

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Therapeutic potential of appropriately evaluated safe-induced pluripotent stem cells for spinal cord injury

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 107 （ 28 ） 12704 - 12709 2010年07月

研究論文（学術雑誌）, ISSN 0027-8424

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Regulation of adult neural progenitor cells by Galectin-1/beta 1 Integrin interaction

岡野栄之

JOURNAL OF NEUROCHEMISTRY 113 （ 6 ） 1516 - 1524 2010年06月

研究論文（学術雑誌）, ISSN 0022-3042

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Lizards respond to an extremely low-frequency electromagnetic field

岡野栄之

JOURNAL OF EXPERIMENTAL BIOLOGY 213 （ 12 ） 1985 - 1990 2010年06月

研究論文（学術雑誌）, ISSN 0022-0949

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Synaptic Input of ON-Bipolar Cells onto the Dopaminergic Neurons of the Mouse Retina

岡野栄之

JOURNAL OF COMPARATIVE NEUROLOGY 518 （ 11 ） 2035 - 2050 2010年06月

研究論文（学術雑誌）, ISSN 0021-9967

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The Musashi family RNA-binding proteins in stem cells

Horisawa Kenichi, Imai Takao, Okano Hideyuki, Yanagawa Hiroshi

Biomolecular Concepts 1 （ 1 ） 59 - 66 2010年05月

ISSN 1868-5021

The Musashi family is an evolutionarily conserved group of RNA-binding proteins. In mammal, two members of the group, Msi1 and Msi2, have been identified to date. Msi1 is considered to play roles in maintaining the stem cell status (stemness) of neural stem/progenitor cells in adults and in the development of central nervous system through translational regulation of its target mRNAs, which encode regulators of signal transduction and the cell cycle. Recently, strong expression of Msi1 in various somatic stem/progenitor cells of adult tissues, such as eye, gut, stomach, breast, and hair follicle, has been reported. The protein is also expressed in various cancer cells, and ectopically emerging cells have been found in neural tissues of patients with diseases involving neural disorder, including epilepsy. Many novel target mRNAs and regulatory pathways of Msi1 have been reported in recent years. Here, we present a review of the functions and action mechanisms of Msi1 protein and discuss possible directions for further study.

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The LTB4-BLT1 Axis Mediates Neutrophil Infiltration and Secondary Injury in Experimental Spinal Cord Injury

岡野栄之

AMERICAN JOURNAL OF PATHOLOGY 176 （ 5 ） 2352 - 2366 2010年05月

研究論文（学術雑誌）, ISSN 0002-9440

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Transplantation of Galectin-1-Expressing Human Neural Stem Cells Into the Injured Spinal Cord of Adult Common Marmosets

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 88 （ 7 ） 1394 - 1405 2010年05月

研究論文（学術雑誌）, ISSN 0360-4012

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Analysis of the Expression and Function of BRINP Family Genes During Neuronal Differentiation in Mouse Embryonic Stem Cell-Derived Neural Stem Cells

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 88 （ 7 ） 1387 - 1393 2010年05月

研究論文（学術雑誌）, ISSN 0360-4012

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von Hippel-Lindau protein regulates transition from the fetal to the adult circulatory system in retina

岡野栄之

DEVELOPMENT 137 （ 9 ） 1563 - 1571 2010年05月

研究論文（学術雑誌）, ISSN 0950-1991

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BMP-induced REST regulates the establishment and maintenance of astrocytic identity

岡野栄之

JOURNAL OF CELL BIOLOGY 189 （ 1 ） 159 - U220 2010年04月

研究論文（学術雑誌）, ISSN 0021-9525

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Musashi1 as a marker of reactive astrocytes after transient focal brain ischemia

岡野栄之

NEUROSCIENCE RESEARCH 66 （ 4 ） 390 - 395 2010年04月

研究論文（学術雑誌）, ISSN 0168-0102

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Regional Spinal Cord Cooling Using a Countercurrent Closed-Lumen Epidural Catheter

岡野栄之

ANNALS OF THORACIC SURGERY 89 （ 4 ） 1312 - 1313 2010年04月

研究論文（学術雑誌）, ISSN 0003-4975

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Neural stem cells and strategies for the regeneration of the central nervous system

岡野栄之

PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES 86 （ 4 ） 438 - 450 2010年04月

研究論文（学術雑誌）, ISSN 0386-2208

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Blockade of interleukin-6 signaling suppressed cochlear inflammatory response and improved hearing impairment in noise-damaged mice cochlea

岡野栄之

NEUROSCIENCE RESEARCH 66 （ 4 ） 345 - 352 2010年04月

研究論文（学術雑誌）, ISSN 0168-0102

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Stem Cell-Like Properties of the Endometrial Side Population: Implication in Endometrial Regeneration

岡野栄之

PLOS ONE 5 （ 4 ） E161 - E161 2010年04月

研究論文（学術雑誌）, ISSN 1932-6203

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Subventricular Zone-Derived Neural Progenitor Cells Migrate Along a Blood Vessel Scaffold Toward the Post-stroke Striatum

岡野栄之

STEM CELLS 28 （ 3 ） 545 - 554 2010年03月

研究論文（学術雑誌）, ISSN 1066-5099

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Stromal Cell-Secreted Factors Promote the Survival of Embryonic Stem Cell-Derived Early Neural Stem/Progenitor Cells Via the Activation of MAPK and PI3K-Akt Pathways

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 88 （ 4 ） 722 - 734 2010年03月

研究論文（学術雑誌）, ISSN 0360-4012

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Seizure suppression in amygdala-kindled mice by transplantation of neural stem/progenitor cells derived from mouse embryonic stem cells

Shindo Atsushi, Nakamura Takehiro, Matsumoto Yoshihito, Kawai Nobuyuki, Okano Hideyuki, Nagao Seigo, Itano Toshifumi, Tamiya Takashi

Neurologia Medico-Chirurgica 50 （ 2 ） 98 - 105 2010年02月

ISSN 0470-8105

Embryonic stem cells (ES cells) differentiate into multiple cell lineages including neural cells. The present study optimized the method to induce differentiation of gamma-aminobutyric acid-producing neurons (GABAergic neurons) from ES cell-derived neural stem/progenitor cells (NS/PCs), and transplanted these ES cell-derived GABAergic neurons producing neural progenitors into kindled epileptic mice, and analyzed the morphological and functional recovery from epilepsy. The response of kindling was evaluated by the modified Racine scale. Following stage 5 kindling, the mice were divided into two groups. Group 1 received NS/PCs derived from the ES cells ubiquitously expressing green fluorescent protein transplanted into the dorsal hippocampal area. Group 2 received microinjections of only the medium. After transplantation, the recovery of seizures was evaluated by the modified Racine scale again. All mice were perfused and fixed for immunohistochemical analysis after finishing the kindling experiment. In Group 1, one mouse was classified as stage 0, five as stage 3, and one as stage 4 recovering from stage 5 at 6 weeks after transplantation. In Group 2, all mice remained in stage 5. The transplanted cells were examined immunohistochemically using neuronal and GABAergic markers. In the transplanted mice, substantial hippocampal GABAergic re-innervation and seizure-suppressing effects were observed. NS/PCs derived from ES cells have high potential for use in transplantation therapy for clinically intractable epilepsies.

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Efficient Generation of Mature Cerebellar Purkinje Cells From Mouse Embryonic Stem Cells

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 88 （ 2 ） 234 - 247 2010年02月

研究論文（学術雑誌）, ISSN 0360-4012

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Heart failure causes cholinergic transdifferentiation of cardiac sympathetic nerves via gp130-signaling cytokines in rodents

岡野栄之

JOURNAL OF CLINICAL INVESTIGATION 120 （ 2 ） 408 - 421 2010年02月

研究論文（学術雑誌）, ISSN 0021-9738

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Seizure Suppression in Amygdala-Kindled Mice by Transplantation of Neural Stem/Progenitor Cells Derived From Mouse Embryonic Stem Cells

岡野栄之

NEUROLOGIA MEDICO-CHIRURGICA 50 （ 2 ） 98 - 105 2010年02月

研究論文（学術雑誌）, ISSN 0470-8105

Fbxo45, a Novel Ubiquitin Ligase, Regulates Synaptic Activity

岡野栄之

JOURNAL OF BIOLOGICAL CHEMISTRY 285 （ 6 ） 3840 - 3849 2010年02月

研究論文（学術雑誌）, ISSN 0021-9258

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Molecular Identity of Periglomerular and Short Axon Cells

岡野栄之

JOURNAL OF NEUROSCIENCE 30 （ 3 ） 1185 - 1196 2010年01月

研究論文（学術雑誌）, ISSN 0270-6474

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Spatial Gradient Effects of 120 mT Static Magnetic Field on Endothelial Tubular Formation In Vitro (vol 29, pg 233, 2008)

岡野栄之

BIOELECTROMAGNETICS 31 （ 1 ） 84 - 84 2010年01月

研究論文（学術雑誌）, ISSN 0197-8462

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Investigation of regeneration of damaged CNS and neurological diseases mechanisms using iPS cells

Okano Hideyuki

Clinical Neurology 50 （ 11 ） 887 2010年

ISSN 0009-918X

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Significance of remyelination in the functional recovery after transplantation of NSPCs to SCI

岡野栄之

NEUROSCIENCE RESEARCH 68 E359 - E359 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Induction of neural crest cells from mouse embryonic stem cells in a serum-free monolayer culture

岡野栄之

INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY 54 （ 8-9 ） 1287 - 1294 2010年

研究論文（学術雑誌）, ISSN 0214-6282

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Protein transduction of p38 MAP kinase enhances adult neural stem cell migration.

岡野栄之

NEUROSCIENCE RESEARCH 68 E254 - E254 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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The maintenance of Purkinje axons by HuC in adult cerebellum

岡野栄之

NEUROSCIENCE RESEARCH 68 E61 - E61 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Transplantation of human iPS cell-derived neurospheres for the treatment of spinal cord injury in NOD-scid mice

岡野栄之

NEUROSCIENCE RESEARCH 68 E427 - E427 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Strategies toward CNS-Regeneration using iPS cell technology

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 112 28P - 28P 2010年

研究論文（学術雑誌）, ISSN 1347-8613

Multiple expression analysis of neurotransmittor under embryonic stem cell differentiation

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 112 28P - 28P 2010年

研究論文（学術雑誌）, ISSN 1347-8613

Investigating the roles of purinergic signaling in adult neurogenesis

岡野栄之

NEUROSCIENCE RESEARCH 68 E367 - E367 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Drug discovery using human neurons derived from iPS cells

岡野栄之

NEUROSCIENCE RESEARCH 68 E134 - E135 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Fbxo45, a novel ubiquitin ligase, regulates synaptic activity

岡野栄之

NEUROSCIENCE RESEARCH 68 E60 - E60 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Combining extensive treadmill training with a selective semaphorin3A inhibitor treatment enhances locomotor functional recovery by wiring regenerated axons in adult spinal cord-transected rats

岡野栄之

NEUROSCIENCE RESEARCH 68 E259 - E260 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Development and analysis of less invasive transient middle cerebral artery occlusion model on Common Marmoset

岡野栄之

NEUROSCIENCE RESEARCH 68 E320 - E320 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Galectin-1 is expressed in early type neural progenitor cells and down-regulates neurogenesis in the adult hippocampus

岡野栄之

NEUROSCIENCE RESEARCH 68 E365 - E365 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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In silico analysis of hnRNP K binding sites in p21 mRNA

岡野栄之

NEUROSCIENCE RESEARCH 68 E354 - E354 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Planar cell polarity of multiciliated ependymal cells regulated by non-muscle myosin II

岡野栄之

NEUROSCIENCE RESEARCH 68 E364 - E364 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Role of G-protein coupled receptors in the neural differentiation from embryonic stem cells

岡野栄之

NEUROSCIENCE RESEARCH 68 E134 - E134 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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New neurons form and maintain their path of astrocytic processes for rapid migration in the adult brain

岡野栄之

NEUROSCIENCE RESEARCH 68 E93 - E93 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Implication of monoamines and neuropeptides in embryogenesisis and neurogenesis

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 112 70P - 70P 2010年

研究論文（学術雑誌）, ISSN 1347-8613

Transplantation of human embryonic stem cell derived-neural stem/progenitor cells for spinal cord injury in adult NOD/Scid mice

岡野栄之

NEUROSCIENCE RESEARCH 68 E425 - E425 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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"Color Timer" mice: visualization of neuronal differentiation with fluorescent proteins

岡野栄之

MOLECULAR BRAIN 3 S230 - S230 2010年

研究論文（学術雑誌）, ISSN 1756-6606

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Generation of induced pluripotent stem cells from patients with schizophrenia

岡野栄之

NEUROSCIENCE RESEARCH 68 E314 - E314 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Analysis of molecular mechanism underlying the control of ES cell differentiation by the stimulation of dopamine receptors located on embryonic stem cells

岡野栄之

NEUROSCIENCE RESEARCH 68 E134 - E134 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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New aspects on the migration and differentiation of neural crest stem cells

岡野栄之

NEUROSCIENCE RESEARCH 68 E39 - E39 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Strategies toward regeneration of damaged CNS using iPS cells

岡野栄之

NEUROSCIENCE RESEARCH 68 E53 - E53 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Agenetic approach for the organization and function of the blood brain barrier in Drosophila

岡野栄之

NEUROSCIENCE RESEARCH 68 E57 - E57 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Functional analysis of macrophage migration inhibitory factor (MIF) in the mouse neural stem/progenitor cells

岡野栄之

NEUROSCIENCE RESEARCH 68 E245 - E246 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Touch and pain receptive network formation with RNA binding protein Musashi2 and Pleiotrophin

岡野栄之

NEUROSCIENCE RESEARCH 68 E161 - E161 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Comparison of the neural crest stem cell characteristics derived from dorsal root ganglia, whisker pad and bone marrow in neonate rodents

岡野栄之

NEUROSCIENCE RESEARCH 68 E129 - E129 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Role and mechanisms of reactive astrocytes? Migration after spinal cord injury

岡野栄之

NEUROSCIENCE RESEARCH 68 E242 - E243 2010年

研究論文（学術雑誌）, ISSN 0168-0102

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Sox10-Venus mice: a new tool for real-time labeling of neural crest lineage cells and oligodendrocytes

岡野栄之

MOLECULAR BRAIN 3 2127 - 2128 2010年

研究論文（学術雑誌）, ISSN 1756-6606

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The common marmoset as a model system for neuroscience

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 60 S28 - S28 2010年

研究論文（学術雑誌）, ISSN 1880-6546

Modeling familial Alzheimer's disease with induced pluripotent stem cells

岡野栄之

HUMAN MOLECULAR GENETICS 20 （ 23 ） 4530 - 4539 2011年12月

研究論文（学術雑誌）, ISSN 0964-6906

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Generation of Stratified Squamous Epithelial Progenitor Cells from Mouse Induced Pluripotent Stem Cells

岡野栄之

PLOS ONE 6 （ 12 ） S89 - S89 2011年12月

研究論文（学術雑誌）, ISSN 1932-6203

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Significance of Remyelination by Neural Stem/Progenitor Cells Transplanted into the Injured Spinal Cord

岡野栄之

STEM CELLS 29 （ 12 ） 1983 - 1994 2011年12月

研究論文（学術雑誌）, ISSN 1066-5099

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Host-derived MMP-13 exhibits a protective role in lung metastasis of melanoma cells by local endostatin production

Fukuda H., Mochizuki S., Abe H., Okano H. J., Hara-Miyauchi C., Okano H., Yamaguchi N., Nakayama M., D'Armiento J., Okada Y.

British Journal of Cancer 105 （ 10 ） 1615 - 1624 2011年11月

ISSN 0007-0920

Background: Although matrix metalloproteinases (MMPs) are implicated in tumourigenesis and cancer progression, the role of MMP-13 in melanoma cell metastases is poorly understood. Methods: Lung metastases of mouse melanoma B16BL6 cells were analysed in MMP-13 knockout (KO) and wild-type (WT) mice after intravenous injection. The mRNA and protein expression of MMP-13 in lung tissues was analysed by RT-PCR, real-time PCR, immunoblotting and immunohistochemistry. The expression of SDF-1α, CXCR4 and endostatin, and effects of endostatin to cultured melanoma cells and lung metastases were also studied. Results: Lung metastases of B16BL6 cells were significantly higher by 2.5-5.7-fold in MMP-13 KO mice than in WT mice. The expression of MMP-13 in WT mouse lung tissue was stimulated on day 1 after intravenous injection of the melanoma cells and MMP-13 was immunolocalised to vascular endothelial cells in the lungs. Endostatin formation, but not degradation of SDF-1α, in the lung tissue was associated with reduced lung metastasis in WT mice. Endostatin significantly inhibited migration of B16BL6 cells in monolayer wounding assay and remarkably suppressed Matrigel invasion and transendothelial invasion of the cells. In addition, lung metastases of melanoma cells in MMP-13 KO mice were reduced by intraperitoneal administration of endostatin. Conclusion: Our results suggest that MMP-13 is overproduced by endothelial cells in the lungs with melanoma cells and has a protective role in lung metastasis by local generation of endostatin. © 2011 Cancer Research UK All rights reserved.

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Human Hepatocyte Growth Factor Promotes Functional Recovery in Primates after Spinal Cord Injury

岡野栄之

PLOS ONE 6 （ 11 ） 3521 - 3534 2011年11月

研究論文（学術雑誌）, ISSN 1932-6203

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IN VITRO CHEMOSENSITIVITY OF HUMAN GLIOMA STEM CELLS COMPARED WITH THAT OF NEURAL STEM CELLS FROM HUMAN IPS CELLS

岡野栄之

NEURO-ONCOLOGY 13 146 - 146 2011年11月

研究論文（学術雑誌）, ISSN 1522-8517

Conserved metabolic energy production pathways govern Eiger/TNF-induced nonapoptotic cell death

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 108 （ 47 ） 18977 - 18982 2011年11月

研究論文（学術雑誌）, ISSN 0027-8424

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Beneficial compaction of spinal cord lesion by migrating astrocytes through glycogen synthase kinase-3 inhibition

岡野栄之

EMBO MOLECULAR MEDICINE 3 （ 11 ） 682 - 696 2011年11月

研究論文（学術雑誌）, ISSN 1757-4676

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Stress-Activated Protein Kinase MKK7 Regulates Axon Elongation in the Developing Cerebral Cortex

岡野栄之

JOURNAL OF NEUROSCIENCE 31 （ 46 ） 16872 - 16883 2011年11月

研究論文（学術雑誌）, ISSN 0270-6474

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Strategic approaches to regeneration of a damaged central nervous system

Okano Hideyuki

Cornea 30 （ 10 SUPPL. 1 ） 2011年10月

ISSN 0277-3740

It has long been believed that a damaged central nervous system (CNS) is difficult to regenerate; however, as a result of the rapid progress in biomedical science, including stem cell biology, there has been a renewed and an increasing interest in developing regenerative therapies for the CNS. Driving this interest has been the discovery of neural stem cells in the adult mammalian CNS, including the human CNS, and the finding that neurogenesis occurs in the adult brain. To achieve regeneration of a damaged CNS, our main strategies encompass: (1) axonal regeneration, (2) replenishment of lost neural cells, and (3) functional recovery. In this review, achievements using these approaches will be outlined. © 2011 by Lippincott Williams & Wilkins.

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Grafted human-induced pluripotent stem-cell-derived neurospheres promote motor functional recovery after spinal cord injury in mice

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 108 （ 40 ） 16825 - 16830 2011年10月

研究論文（学術雑誌）, ISSN 0027-8424

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Dysfunction of fibroblasts of extrarenal origin underlies renal fibrosis and renal anemia in mice

岡野栄之

JOURNAL OF CLINICAL INVESTIGATION 121 （ 10 ） 3981 - 3990 2011年10月

研究論文（学術雑誌）, ISSN 0021-9738

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Cell Therapy for Spinal Cord Injury by Neural Stem/Progenitor Cells Derived from iPS/ES Cells

岡野栄之

NEUROTHERAPEUTICS 8 （ 4 ） 668 - 676 2011年10月

研究論文（学術雑誌）, ISSN 1933-7213

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Strategic Approaches to Regeneration of a Damaged Central Nervous System

岡野栄之

CORNEA 30 S15 - S18 2011年10月

研究論文（学術雑誌）, ISSN 0277-3740

Transplantation of human neural stem/progenitor cells overexpressing galectin-1 improves functional recovery from focal brain ischemia in the mongolian gerbil

岡野栄之

MOLECULAR BRAIN 4 739 - 750 2011年09月

研究論文（学術雑誌）, ISSN 1756-6606

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Impaired spatial and contextual memory formation in galectin-1 deficient mice

岡野栄之

MOLECULAR BRAIN 4 170 - 170 2011年09月

研究論文（学術雑誌）, ISSN 1756-6606

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The dual origin of the peripheral olfactory system: placode and neural crest

岡野栄之

MOLECULAR BRAIN 4 103 - 113 2011年09月

研究論文（学術雑誌）, ISSN 1756-6606

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Gene Targeting and Subsequent Site-Specific Transgenesis at the beta-actin (ACTB) Locus in Common Marmoset Embryonic Stem Cells

岡野栄之

STEM CELLS AND DEVELOPMENT 20 （ 9 ） 1588 - 1600 2011年09月

研究論文（学術雑誌）, ISSN 1547-3287

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An efficient method to generate human iPS-derived epithelial progenitors/keratinocyte

岡野栄之

JOURNAL OF INVESTIGATIVE DERMATOLOGY 131 S77 - S77 2011年09月

研究論文（学術雑誌）, ISSN 0022-202X

Special Issue in Honor of Professor Katsuhiko Mikoshiba

岡野栄之

NEUROCHEMICAL RESEARCH 36 （ 7 ） 1147 - 1148 2011年07月

研究論文（学術雑誌）, ISSN 0364-3190

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Expression and Function of Sox21 During Mouse Cochlea Development

岡野栄之

NEUROCHEMICAL RESEARCH 36 （ 7 ） 1261 - 1269 2011年07月

研究論文（学術雑誌）, ISSN 0364-3190

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Mimicking the Neurotrophic Factor Profile of Embryonic Spinal Cord Controls the Differentiation Potential of Spinal Progenitors into Neuronal Cells

岡野栄之

PLOS ONE 6 （ 6 ） 33 - 41 2011年06月

研究論文（学術雑誌）, ISSN 1932-6203

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Generating mouse models of degenerative diseases using Ore/lox-mediated in vivo mosaic cell ablation

岡野栄之

JOURNAL OF CLINICAL INVESTIGATION 121 （ 6 ） 2462 - 2469 2011年06月

研究論文（学術雑誌）, ISSN 0021-9738

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Schwann-Spheres Derived from Injured Peripheral Nerves in Adult Mice - Their In Vitro Characterization and Therapeutic Potential

岡野栄之

PLOS ONE 6 （ 6 ） 515 - 518 2011年06月

研究論文（学術雑誌）, ISSN 1932-6203

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Cell Surface N-Glycans-Mediated Isolation of Mouse Neural Stem Cells

岡野栄之

JOURNAL OF CELLULAR PHYSIOLOGY 226 （ 6 ） 1433 - 1438 2011年06月

研究論文（学術雑誌）, ISSN 0021-9541

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Calcium signal regulates temperature-dependent transformation of sporozoites in malaria parasite development

岡野栄之

EXPERIMENTAL PARASITOLOGY 128 （ 2 ） 176 - 180 2011年06月

研究論文（学術雑誌）, ISSN 0014-4894

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Functional analysis of HOXD9 in human gliomas and glioma cancer stem cells

岡野栄之

MOLECULAR CANCER 10 866 - 876 2011年05月

研究論文（学術雑誌）, ISSN 1476-4598

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DISC1-dependent switch from progenitor proliferation to migration in the developing cortex

岡野栄之

NATURE 473 （ 7345 ） 92 - U107 2011年05月

研究論文（学術雑誌）, ISSN 0028-0836

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Musashi1 Cooperates in Abnormal Cell Lineage Protein 28 (Lin28)-mediated Let-7 Family MicroRNA Biogenesis in Early Neural Differentiation

岡野栄之

JOURNAL OF BIOLOGICAL CHEMISTRY 286 （ 18 ） 683 - 688 2011年05月

研究論文（学術雑誌）, ISSN 0021-9258

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Isolation and function of mouse tissue resident vascular precursors marked by myelin protein zero

岡野栄之

JOURNAL OF EXPERIMENTAL MEDICINE 208 （ 5 ） 949 - 960 2011年05月

研究論文（学術雑誌）, ISSN 0022-1007

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Properties of dopaminergic neurons in organotypic mesencephalic-striatal co-cultures - evidence for a facilitatory effect of dopamine on the glutamatergic input mediated by alpha-1 adrenergic receptors

岡野栄之

EUROPEAN JOURNAL OF NEUROSCIENCE 33 （ 9 ） 1622 - 1636 2011年05月

研究論文（学術雑誌）, ISSN 0953-816X

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Schwann Cell Plasticity After Spinal Cord Injury Shown by Neural Crest Lineage Tracing

岡野栄之

GLIA 59 （ 5 ） 771 - 784 2011年05月

研究論文（学術雑誌）, ISSN 0894-1491

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Identification of a novel intronic enhancer responsible for the transcriptional regulation of musashi1 in neural stem/progenitor cells

岡野栄之

MOLECULAR BRAIN 4 1841 - 1853 2011年04月

研究論文（学術雑誌）, ISSN 1756-6606

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Fbxw7-dependent Degradation of Notch Is Required for Control of "Stemness" and Neuronal-Glial Differentiation in Neural Stem Cells

岡野栄之

JOURNAL OF BIOLOGICAL CHEMISTRY 286 （ 15 ） 13754 - 13764 2011年04月

研究論文（学術雑誌）, ISSN 0021-9258

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Neural Crest-Derived Stem Cells Migrate and Differentiate Into Cardiomyocytes After Myocardial Infarction

岡野栄之

ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY 31 （ 3 ） 582 - U234 2011年03月

研究論文（学術雑誌）, ISSN 1079-5642

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Downregulation of Notch mediates the seamless transition of individual Drosophila neuroepithelial progenitors into optic medullar neuroblasts during prolonged G1

岡野栄之

DEVELOPMENTAL BIOLOGY 351 （ 1 ） 163 - 175 2011年03月

研究論文（学術雑誌）, ISSN 0012-1606

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Cellular Composition and Organization of the Subventricular Zone and Rostral Migratory Stream in the Adult and Neonatal Common Marmoset Brain

岡野栄之

JOURNAL OF COMPARATIVE NEUROLOGY 519 （ 4 ） 690 - 713 2011年03月

研究論文（学術雑誌）, ISSN 0021-9967

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Purified Mesenchymal Stem Cells Are an Efficient Source for iPS Cell Induction

岡野栄之

PLOS ONE 6 （ 3 ） S90 - S91 2011年03月

研究論文（学術雑誌）, ISSN 1932-6203

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A 1-mu T extremely low-frequency electromagnetic field vs. sham control for mild-to-moderate hypertension: a double-blind, randomized study

岡野栄之

HYPERTENSION RESEARCH 34 （ 3 ） 372 - 377 2011年03月

研究論文（学術雑誌）, ISSN 0916-9636

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Population-averaged standard template brain atlas for the common marmoset (Callithrix jacchus)

Hikishima K., Quallo M. M., Komaki Y., Yamada M., Kawai K., Momoshima S., Okano H. J., Sasaki E., Tamaoki N., Lemon R. N., Iriki A., Okano H.

NeuroImage 54 （ 4 ） 2741 - 2749 2011年02月

ISSN 1053-8119

Advanced magnetic resonance (MR) neuroimaging analysis techniques based on voxel-wise statistics, such as voxel-based morphometry (VBM) and functional MRI, are widely applied to cognitive brain research in both human subjects and in non-human primates. Recent developments in imaging have enabled the evaluation of smaller animal models with sufficient spatial resolution. The common marmoset (Callithrix jacchus), a small New World primate species, has been widely used in neuroscience research, to which voxel-wise statistics could be extended with a species-specific brain template. Here, we report, for the first time, a tissue-segmented, population-averaged standard template of the common marmoset brain. This template was created by using anatomical T₁-weighted images from 22 adult marmosets with a high-resolution isotropic voxel size of (0.2mm)³ at 7-Tesla and DARTEL algorithm in SPM8. Whole brain templates are available at International Neuroinformatics Japan Node website, http://brainatlas.brain.riken.jp/marmoset/. © 2010 Elsevier Inc.

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Akt Suppresses Retrograde Degeneration of Dopaminergic Axons by Inhibition of Macroautophagy

岡野栄之

JOURNAL OF NEUROSCIENCE 31 （ 6 ） 2125 - 2135 2011年02月

研究論文（学術雑誌）, ISSN 0270-6474

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Generation of Human Melanocytes from Induced Pluripotent Stem Cells

岡野栄之

PLOS ONE 6 （ 1 ） 101P - 101P 2011年01月

研究論文（学術雑誌）, ISSN 1932-6203

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Effect of Magnetotherapeutic Device on Pain Associated With Neck and Shoulder Stiffness

岡野栄之

ALTERNATIVE THERAPIES IN HEALTH AND MEDICINE 17 （ 6 ） 44 - 48 2011年01月

研究論文（学術雑誌）, ISSN 1078-6791

Galectin-1 is expressed in early-type neural progenitor cells and down-regulates neurogenesis in the adult hippocampus

岡野栄之

MOLECULAR BRAIN 4 683 - 688 2011年01月

研究論文（学術雑誌）, ISSN 1756-6606

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Evaluation of human iPS cells by neural differentiation and tumorigenicity

Okada Yohei, Okano Hideyuki

Clinical Neurology 51 （ 11 ） 1079 - 1080 2011年

ISSN 0009-918X

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A vascular niche factor for neural stem cells of the mammalian brain

岡野栄之

NEUROSCIENCE RESEARCH 71 E51 - E51 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Efficient derivation of neural stem cells from common marmoset ES cells and iPS cells

岡野栄之

NEUROSCIENCE RESEARCH 71 E332 - E332 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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In silico analysis of hnRNP K bunding sites in p21 mRNA and its in vitro examination

岡野栄之

NEUROSCIENCE RESEARCH 71 E225 - E225 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Development and analysis of less invasive transient middle cerebral artery occlusion model on non-human primate

岡野栄之

NEUROSCIENCE RESEARCH 71 E403 - E404 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Novel Concept of Motor Functional Analysis for Spinal Cord Injury in Adult Mice

岡野栄之

JOURNAL OF BIOMEDICINE AND BIOTECHNOLOGY 18977 - 18982 2011年

研究論文（学術雑誌）, ISSN 1110-7243

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Application of ES/somatic cell cloning technique to the production of genetically modified marmosets

岡野栄之

NEUROSCIENCE RESEARCH 71 E41 - E41 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Sensory network formation with RNA binding protein Musashi2

岡野栄之

NEUROSCIENCE RESEARCH 71 E225 - E225 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Regulation of Posttranscriptional Modification as a Possible Therapeutic Approach for Retinal Neuroprotection

岡野栄之

JOURNAL OF OPHTHALMOLOGY 1702 - 1710 2011年

研究論文（学術雑誌）, ISSN 2090-004X

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The maintenance of Purkinje axons by an RNA-binding protein HuC in adult cerebellum

岡野栄之

NEUROSCIENCE RESEARCH 71 E95 - E96 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Transgenic technologies in nonhuman primates

岡野栄之

NEUROSCIENCE RESEARCH 71 E41 - E41 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Comparative Study of Methods for Administering Neural Stem/Progenitor Cells to Treat Spinal Cord Injury in Mice

岡野栄之

CELL TRANSPLANTATION 20 （ 5 ） 727 - 739 2011年

研究論文（学術雑誌）, ISSN 0963-6897

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Disrupted-in-Schizoprenia-1 regulates transport of neuronal mRNA for neural plasticity

岡野栄之

NEUROSCIENCE RESEARCH 71 E323 - E323 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Generation and characterization of human ES cells carrying Sox1-reporter gene for neural differentiation

岡野栄之

NEUROSCIENCE RESEARCH 71 E125 - E125 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Direct induction of safe neural stem cells from adult mouse fibroblasts

岡野栄之

NEUROSCIENCE RESEARCH 71 E228 - E228 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Evaluation of human iPS cells by neural differentiation and tumorigenicity

岡野栄之

NEUROSCIENCE RESEARCH 71 E329 - E330 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Dysregulation of NF-kappa B signaling is involved in the pathogenesis of a mouse model for Rett syndrome

岡野栄之

NEUROSCIENCE RESEARCH 71 E401 - E401 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Beneficial compaction of spinal cord lesion by migrating astrocytes through Glycogen Synthase Kinase-3 inhibition

岡野栄之

NEUROSCIENCE RESEARCH 71 E59 - E59 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Multiple analyses of G-protein coupled receptor (GPCR) expression in the neural differentiation from embryonic stem cells

岡野栄之

NEUROSCIENCE RESEARCH 71 E126 - E126 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Recovery Effects of a 180 mT Static Magnetic Field on Bone Mineral Density of Osteoporotic Lumbar Vertebrae in Ovariectomized Rats

岡野栄之

EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE 1 - 8 2011年

研究論文（学術雑誌）, ISSN 1741-427X

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Analysis for gene expression program governing specific neural network formation

岡野栄之

NEUROSCIENCE RESEARCH 71 E336 - E336 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Directional migration of new neurons generated by transplanted human iPS cell-derived neural stem cells in the striatum of a mouse stroke model

岡野栄之

NEUROSCIENCE RESEARCH 71 E338 - E338 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Regeneration of the damaged CNS using human iPSCs-derived neural progenitor cells

岡野栄之

NEUROSCIENCE RESEARCH 71 E26 - E26 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Molecular basis of the integrity of blood-brain barrier in Drosophila

岡野栄之

NEUROSCIENCE RESEARCH 71 E59 - E59 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Eiger and Wengen: The Drosophila Orthologs of TNF/TNFR

岡野栄之

ADVANCES IN TNF FAMILY RESEARCH 691 45 - 50 2011年

研究論文（学術雑誌）, ISSN 0065-2598

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Generation of neural crest progenitor cells from human induced pluripotent stem cells by a simple method

岡野栄之

NEUROSCIENCE RESEARCH 71 E127 - E127 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Efficient generation and developmental analysis of basal forebrain cholinergic neurons from mouse embryonic stem cells

岡野栄之

NEUROSCIENCE RESEARCH 71 E332 - E332 2011年

研究論文（学術雑誌）, ISSN 0168-0102

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Isolation of mouse mesenchymal stem cells on the basis of expression of Sca-1 and PDGFR-α

Houlihan Diarmaid D., Mabuchi Yo, Morikawa Satoru, Niibe Kunimichi, Araki Daisuke, Suzuki Sadafumi, Okano Hideyuki, Matsuzaki Yumi

Nature Protocols 7 （ 12 ） 2103 - 2111 2012年12月

ISSN 1754-2189

Platelet-derived growth factor receptor α (PDGFR-α) and stem cell antigen 1 (Sca-1) have recently been identified as selective markers of mouse mesenchymal stem cells (MSCs). PDGFR-α + Sca-1 + (PαS) MSCs have augmented growth potential and robust tri-lineage differentiation compared with standard culture-selected MSCs. In addition, the selective isolation of PαS MSCs avoids cellular contamination that can complicate other methods. Here we describe in detail our protocol to isolate PαS MSCs using flow cytometry. In brief, the tibia and femora are isolated and crushed using a pestle and mortar. The crushed bones are then chopped and incubated for 1 h at 37°C in 20 ml of DMEM containing 0.2% (wt/vol) collagenase. The cell suspension is filtered before red blood cell lysis and incubated with the following antibodies: allophycocyanin (APC)-conjugated PDGFR-α, FITC-conjugated Sca-1, phycoerythrin (PE)-conjugated CD45 and Ter119. Appropriate gates are constructed on a cell sorter to exclude dead cells and lineage (CD45 + Ter-119 +)-positive cells. Approximately 10,000 PαS MSCs may then be isolated per mouse. The total protocol takes ∼7 h to complete. © 2012 Nature America, Inc.

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Isolation of mouse mesenchymal stem cells on the basis of expression of Sca-1 and PDGFR-alpha

岡野栄之

NATURE PROTOCOLS 7 （ 12 ） 2103 - 2111 2012年12月

研究論文（学術雑誌）, ISSN 1754-2189

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The common marmoset as a novel animal model system for biomedical and neuroscience research applications

岡野栄之

SEMINARS IN FETAL & NEONATAL MEDICINE 17 （ 6 ） 336 - 340 2012年12月

研究論文（学術雑誌）, ISSN 1744-165X

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Balanced ubiquitination determines cellular responsiveness to extracellular stimuli

岡野栄之

CELLULAR AND MOLECULAR LIFE SCIENCES 69 （ 23 ） 4007 - 4016 2012年12月

研究論文（学術雑誌）, ISSN 1420-682X

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Conditions for quantitative evaluation of injured spinal cord by in vivo diffusion tensor imaging and tractography: Preclinical longitudinal study in common marmosets

岡野栄之

NEUROIMAGE 63 （ 4 ） 1841 - 1853 2012年12月

研究論文（学術雑誌）, ISSN 1053-8119

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Comparative gene expression in developing cognitive circuits in common marmoset brain

岡野栄之

INTERNATIONAL JOURNAL OF DEVELOPMENTAL NEUROSCIENCE 30 （ 8 ） 669 - 669 2012年12月

研究論文（学術雑誌）, ISSN 0736-5748

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Novel In Vivo Imaging Analysis of an Inner Ear Drug Delivery System in Mice: Comparison of Inner Ear Drug Concentrations over Time after Transtympanic and Systemic Injections

岡野栄之

PLOS ONE 7 （ 12 ） 163 - 175 2012年12月

研究論文（学術雑誌）, ISSN 1932-6203

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Comparative Anatomy of Marmoset and Mouse Cortex from Genomic Expression (April, pg 5039, 2012)

岡野栄之

JOURNAL OF NEUROSCIENCE 32 （ 49 ） 17932 - 17932 2012年12月

研究論文（学術雑誌）, ISSN 0270-6474

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Pre-Evaluated Safe Human iPSC-Derived Neural Stem Cells Promote Functional Recovery after Spinal Cord Injury in Common Marmoset without Tumorigenicity

岡野栄之

PLOS ONE 7 （ 12 ） 58 - 69 2012年12月

研究論文（学術雑誌）, ISSN 1932-6203

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Exhaustion of nucleus pulposus progenitor cells with ageing and degeneration of the intervertebral disc

岡野栄之

NATURE COMMUNICATIONS 3 1067 - 1080 2012年12月

研究論文（学術雑誌）, ISSN 2041-1723

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MRI Characterization of Paranodal Junction Failure and Related Spinal Cord Changes in Mice

岡野栄之

PLOS ONE 7 （ 12 ） 6106 - 6125 2012年12月

研究論文（学術雑誌）, ISSN 1932-6203

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Efficient Derivation of Multipotent Neural Stem/Progenitor Cells from Non-Human Primate Embryonic Stem Cells

岡野栄之

PLOS ONE 7 （ 11 ） 24561 - 24570 2012年11月

研究論文（学術雑誌）, ISSN 1932-6203

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The Semaphorin 3A Inhibitor SM-345431 Accelerates Peripheral Nerve Regeneration and Sensitivity in a Murine Corneal Transplantation Model

岡野栄之

PLOS ONE 7 （ 11 ） 106 - 114 2012年11月

研究論文（学術雑誌）, ISSN 1932-6203

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Sustained bFGF-Release Tubes for Peripheral Nerve Regeneration: Comparison with Autograft

岡野栄之

PLASTIC AND RECONSTRUCTIVE SURGERY 130 （ 4 ） 866 - 876 2012年10月

研究論文（学術雑誌）, ISSN 0032-1052

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Direct isolation and RNA-seq reveal environment-dependent properties of engrafted neural stem/progenitor cells

岡野栄之

NATURE COMMUNICATIONS 3 2811 - 2815 2012年10月

研究論文（学術雑誌）, ISSN 2041-1723

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Multiple Analyses of G-Protein Coupled Receptor (GPCR) Expression in the Development of Gefitinib-Resistance in Transforming Non-Small-Cell Lung Cancer

岡野栄之

PLOS ONE 7 （ 10 ） S230 - S230 2012年10月

研究論文（学術雑誌）, ISSN 1932-6203

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Identification of mutations in the prostaglandin transporter gene SLCO2A1 and its phenotype-genotype correlation in Japanese patients with pachydermoperiostosis

岡野栄之

JOURNAL OF DERMATOLOGICAL SCIENCE 68 （ 1 ） 36 - 44 2012年10月

研究論文（学術雑誌）, ISSN 0923-1811

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Mitochondrial dysfunction associated with increased oxidative stress and alpha-synuclein accumulation in PARK2 iPSC-derived neurons and postmortem brain tissue

岡野栄之

MOLECULAR BRAIN 5 36 - 44 2012年10月

研究論文（学術雑誌）, ISSN 1756-6606

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Neuronal Elav-like (Hu) Proteins Regulate RNA Splicing and Abundance to Control Glutamate Levels and Neuronal Excitability

岡野栄之

NEURON 75 （ 6 ） 1067 - 1080 2012年09月

研究論文（学術雑誌）, ISSN 0896-6273

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Sox21 Promotes Hippocampal Adult Neurogenesis via the Transcriptional Repression of the Hes5 Gene

岡野栄之

JOURNAL OF NEUROSCIENCE 32 （ 36 ） 12543 - 12557 2012年09月

研究論文（学術雑誌）, ISSN 0270-6474

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REGENERATION OF THE DAMAGED CNS USING HUMAN iPSCs-DERIVED NEURAL PROGENITOR CELLS

岡野栄之

ALCOHOLISM-CLINICAL AND EXPERIMENTAL RESEARCH 36 12A - 12A 2012年09月

研究論文（学術雑誌）, ISSN 0145-6008

The effects of moderate-intensity gradient static magnetic fields on nerve conduction

岡野栄之

BIOELECTROMAGNETICS 33 （ 6 ） 518 - 526 2012年09月

研究論文（学術雑誌）, ISSN 0197-8462

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Accelerating progress in induced pluripotent stem cell research for neurological diseases

岡野栄之

ANNALS OF NEUROLOGY 72 （ 2 ） 167 - 174 2012年08月

研究論文（学術雑誌）, ISSN 0364-5134

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Roles of Planar Cell Polarity Signaling in Maturation of Neuronal Precursor Cells in the Postnatal Mouse Olfactory Bulb

岡野栄之

STEM CELLS 30 （ 8 ） 1726 - 1733 2012年08月

研究論文（学術雑誌）, ISSN 1066-5099

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Cell transplantation for spinal cord injury focusing on iPSCs

岡野栄之

EXPERT OPINION ON BIOLOGICAL THERAPY 12 （ 7 ） 811 - 821 2012年07月

研究論文（学術雑誌）, ISSN 1471-2598

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Purinergic Signaling Promotes Proliferation of Adult Mouse Subventricular Zone Cells

岡野栄之

JOURNAL OF NEUROSCIENCE 32 （ 27 ） 9238 - 9247 2012年07月

研究論文（学術雑誌）, ISSN 0270-6474

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Establishment of Induced Pluripotent Stem Cells from Centenarians for Neurodegenerative Disease Research

岡野栄之

PLOS ONE 7 （ 7 ） 683 - 688 2012年07月

研究論文（学術雑誌）, ISSN 1932-6203

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Epigenetic Regulation of Cancer Stem Cell Genes in Triple-Negative Breast Cancer

岡野栄之

AMERICAN JOURNAL OF PATHOLOGY 181 （ 1 ） 257 - 267 2012年07月

研究論文（学術雑誌）, ISSN 0002-9440

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Macrophage migration inhibitory factor (MIF) promotes cell survival and proliferation of neural stem/progenitor cells

岡野栄之

JOURNAL OF CELL SCIENCE 125 （ 13 ） 3210 - 3220 2012年07月

研究論文（学術雑誌）, ISSN 0021-9533

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Neural stem cells, adult neurogenesis, and galectin-1: From bench to bedside

岡野栄之

DEVELOPMENTAL NEUROBIOLOGY 72 （ 7 ） 1059 - 1067 2012年07月

研究論文（学術雑誌）, ISSN 1932-8451

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Characterization of the RNA-binding protein Musashi1 in zebrafish

岡野栄之

BRAIN RESEARCH 1462 162 - 173 2012年06月

研究論文（学術雑誌）, ISSN 0006-8993

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Xenotransplanted Embryonic Kidney Provides a Niche for Endogenous Mesenchymal Stem Cell Differentiation into Erythropoietin-Producing Tissue

岡野栄之

STEM CELLS 30 （ 6 ） 1228 - 1235 2012年06月

研究論文（学術雑誌）, ISSN 1066-5099

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Neural Stem Cells Directly Differentiated from Partially Reprogrammed Fibroblasts Rapidly Acquire Gliogenic Competency

岡野栄之

STEM CELLS 30 （ 6 ） 1109 - 1119 2012年06月

研究論文（学術雑誌）, ISSN 1066-5099

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Effect of ADAM28 on Carcinoma Cell Metastasis by Cleavage of von Willebrand Factor

岡野栄之

JOURNAL OF THE NATIONAL CANCER INSTITUTE 104 （ 12 ） 906 - 922 2012年06月

研究論文（学術雑誌）, ISSN 0027-8874

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Molecular signatures to define spermatogenic cells in common marmoset (Callithrix jacchus)

岡野栄之

REPRODUCTION 143 （ 5 ） 597 - 609 2012年05月

研究論文（学術雑誌）, ISSN 1470-1626

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Structure of Musashi1 in a complex with target RNA: the role of aromatic stacking interactions

岡野栄之

NUCLEIC ACIDS RESEARCH 40 （ 7 ） 3218 - 3231 2012年04月

研究論文（学術雑誌）, ISSN 0305-1048

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Comparative Anatomy of Marmoset and Mouse Cortex from Genomic Expression

岡野栄之

JOURNAL OF NEUROSCIENCE 32 （ 15 ） 5039 - 5053 2012年04月

研究論文（学術雑誌）, ISSN 0270-6474

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Derivation of Human Differential Photoreceptor-like Cells from the Iris by Defined Combinations of CRX, RX and NEUROD

岡野栄之

PLOS ONE 7 （ 4 ） S230 - S230 2012年04月

研究論文（学術雑誌）, ISSN 1932-6203

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Effect of-opioid receptor agonist on the growth of non-small cell lung cancer (NSCLC) cells

Kuzumaki N., Suzuki A., Narita M., Hosoya T., Nagasawa A., Imai S., Yamamizu K., Morita H., Nagase H., Okada Y., Okano H. J., Yamashita J. K., Okano H., Suzuki T., Narita M.

British Journal of Cancer 106 （ 6 ） 1148 - 1152 2012年03月

ISSN 0007-0920

Background: It is becoming increasingly recognised that opioids are responsible for tumour growth. However, the effects of opioids on tumour growth have been controversial.Methods:The effects of-opioid receptor (KOR) agonist on the growth of non-small cell lung cancer (NSCLC) cells were assessed by a cell proliferation assay. Western blotting was performed to ascertain the mechanism by which treatment with KOR agonist suppresses tumour growth. Results: Addition of the selective KOR agonist U50,488H to gefitinib-sensitive (HCC827) and gefitinib-resistant (H1975) NSCLC cells produced a concentration-dependent decrease in their growth. These effects were abolished by co-treatment with the selective KOR antagonist nor-BNI. Furthermore, the growth-inhibitory effect of gefitinib in HCC827 cells was further enhanced by co-treatment with U50,488H. With regard to the inhibition of tumour growth, the addition of U50, 488H to H1975 cells produced a concentration-dependent decrease in phosphorylated- glycogen synthase kinase 3Β (p-GSK3Β).Conclusion:The present results showed that stimulation of KOR reduces the growth of gefitinib-resistant NSCLC cells through the activation of GSK3Β. © 2012 Cancer Research UK All rights reserved.

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The first clinical trial in Tohoku university hospital after the Great East JAPAN Earthquake

Okano Hideyuki

Keio Journal of Medicine 61 （ 1 ） 3 - 9 2012年03月

ISSN 0022-9717

The Great East Japan Earthquake of 2011 seriously jeopardized our collaborative research with Professor Masashi Aoki (Tohoku University School of Medicine) on the development of new therapies for amyotrophic lateral sclerosis (ALS) using hepatocyte growth factor. After the earthquake struck, Professor Aoki made a tremendous contribution to saving patients' lives and to recovering from the disastrous situation. Thanks to his strong leadership and support from many reliable colleagues, we could finally start new clinical trials for ALS patients. In this article, I wish to introduce Professor Aoki's heroic efforts. © 2012 by The Keio Journal of Medicine.

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The formation of an angiogenic astrocyte template is regulated by the neuroretina in a HIF-1-dependent manner

岡野栄之

DEVELOPMENTAL BIOLOGY 363 （ 1 ） 106 - 114 2012年03月

研究論文（学術雑誌）, ISSN 0012-1606

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RNA-Binding Protein Musashi1 Modulates Glioma Cell Growth through the Post-Transcriptional Regulation of Notch and PI3 Kinase/Akt Signaling Pathways

岡野栄之

PLOS ONE 7 （ 3 ） 1223 - 1237 2012年03月

研究論文（学術雑誌）, ISSN 1932-6203

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RNA-Binding Protein HuD Controls Insulin Translation

岡野栄之

MOLECULAR CELL 45 （ 6 ） 826 - 835 2012年03月

研究論文（学術雑誌）, ISSN 1097-2765

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Identification of Proteasome Components Required for Apical Localization of Chaoptin Using Functional Genomics

岡野栄之

JOURNAL OF NEUROGENETICS 26 （ 1 ） 53 - 63 2012年03月

研究論文（学術雑誌）, ISSN 0167-7063

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Bioluminescent system for dynamic imaging of cell and animal behavior

岡野栄之

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 419 （ 2 ） 188 - 193 2012年03月

研究論文（学術雑誌）, ISSN 0006-291X

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Structure, function and formation of glycans in Drosophila

Yamamoto-Hino Miki, Okano Hideyuki, Kanie Osamu, Goto Satoshi

165 - 188 2012年02月

Glycosylation of proteins shows two striking features: first, there are a large number of glycan structures that are involved in various biological events (diversity) and, second, certain structures are selectively added to specific sites within proteins (specificity) in a context-dependent manner. However, the mechanisms that regulate diversity and specificity are unknown. Over the last few decades, genetic studies in Drosophila have been used to examine the functions mediated by glycosylation in vivo. Recent improvements in the combined use of biochemical/biophysical, genetic, and cell biology approaches has allowed the rapid and detailed analysis of the structure, function and formation of glycans in Drosophila. This chapter aims to provide an overview of the different types of glycans and their biological function(s) in Drosophila. The regulation of glycan synthesis in Drosophila and mammals is also discussed. © 2012 by Nova Science Publishers, Inc. All rights reserved.

Abundant Occurrence of Basal Radial Glia in the Subventricular Zone of Embryonic Neocortex of a Lissencephalic Primate, the Common Marmoset Callithrix jacchus

岡野栄之

CEREBRAL CORTEX 22 （ 2 ） 469 - 481 2012年02月

研究論文（学術雑誌）, ISSN 1047-3211

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Therapeutic potential of induced pluripotent stem cells for spinal cord injury

Nori Satoshi, Tsuji Osahiko, Okada Yohei, Toyama Yoshiaki, Okano Hideyuki, Nakamura Masaya

Brain and Nerve 64 （ 1 ） 17 - 27 2012年01月

ISSN 1881-6096

Once the safety issue has been overcome, induced pluripotent stem cells (iPSCs), which do not entail ethical or immunological concerns, may become the preferred cell source for regenerative medicine. Various types of iPSCs have been established by different methods, and each type exhibits different biological properties. Before iPSC-based clinical applications can be initiated, detailed evaluations of the cells, including their differentiation potentials and tumorigenic activities in different contexts, should be investigated to establish their safety and effectiveness for cell transplantation therapies. Recently, we demonstrated the directed neural differentiation of mouse iPSCs and examined their therapeutic potential in a mouse spinal cord injury (SCI) model. Mouse iPSC-derived neural stem/progenitor cells (NS/PCs), which had been pre-evaluated as non-tumorigenic by their transplantation into nonobese diabetic-severe combined immunodeficiency (NOD-scid) mouse brain, were transplanted into the spinal cord 9 days after SCI. Mouse iPSC-derived NS/PCs differentiated into all three neural lineages without forming teratomas or other tumors. They also participated in re-myelination and induced the axonal re-growth, promoting motor functional recovery. Nevertheless, our results constitute only the first step toward clinical application. The safety and effectiveness of human iPSC-derived NS/PCs need to be more intensively investigated in future preclinical studies, for example, using non-human primate SCI models. In particular, human iPSCs established by delivering reprogramming factors using a safer method than retrovirus system, such as an integration-free virus system, virus-free system, or transgene-free system should be evaluated.

Derivation of human differential photoreceptor-like cells from the iris by defined combinations of CRX, RX and NEUROD.

Seko Yuko, Azuma Noriyuki, Kaneda Makoto, Nakatani Kei, Miyagawa Yoshitaka, Noshiro Yuuki, Kurokawa Reiko, Okano Hideyuki, Umezawa Akihiro

PLoS ONE 7 （ 4 ） 2012年

Examples of direct differentiation by defined transcription factors have been provided for beta-cells, cardiomyocytes and neurons. In the human visual system, there are four kinds of photoreceptors in the retina. Neural retina and iris-pigmented epithelium (IPE) share a common developmental origin, leading us to test whether human iris cells could differentiate to retinal neurons. We here define the transcription factor combinations that can determine human photoreceptor cell fate. Expression of rhodopsin, blue opsin and green/red opsin in induced photoreceptor cells were dependent on combinations of transcription factors: A combination of CRX and NEUROD induced rhodopsin and blue opsin, but did not induce green opsin; a combination of CRX and RX induced blue opsin and green/red opsin, but did not induce rhodopsin. Phototransduction-related genes as well as opsin genes were up-regulated in those cells. Functional analysis; i.e. patch clamp recordings, clearly revealed that generated photoreceptor cells, induced by CRX, RX and NEUROD, responded to light. The response was an inward current instead of the typical outward current. These data suggest that photosensitive photoreceptor cells can be generated by combinations of transcription factors. The combination of CRX and RX generate immature photoreceptors: and additional NEUROD promotes maturation. These findings contribute substantially to a major advance toward eventual cell-based therapy for retinal degenerative diseases.

Nestin Protein Is Phosphorylated in Adult Neural Stem/Progenitor Cells and Not Endothelial Progenitor Cells

岡野栄之

STEM CELLS INTERNATIONAL 1410 - 1422 2012年

研究論文（学術雑誌）, ISSN 1687-966X

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Role of adrenergic receptors in the neural differentiation from pluripotent stem cells

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 118 172P - 172P 2012年

研究論文（学術雑誌）, ISSN 1347-8613

Cisterna-specific Localization of Glycosylation-related Proteins to the Golgi Apparatus

岡野栄之

CELL STRUCTURE AND FUNCTION 37 （ 1 ） 55 - 63 2012年

研究論文（学術雑誌）, ISSN 0386-7196

Control mechanism of gefitinib-resistant human non-small-cell lung cancer (NSCLC) cells growth and transformation by stimulation of delta-opioid receptors

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 118 123P - 123P 2012年

研究論文（学術雑誌）, ISSN 1347-8613

The analysis of telomeric status in "Extraordinary Cancer Resistant Mammal" Naked mole rat

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 118 175P - 175P 2012年

研究論文（学術雑誌）, ISSN 1347-8613

Regeneration of the damaged CNS using human iPSCs-derived neural progenitor cells

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 118 48P - 48P 2012年

研究論文（学術雑誌）, ISSN 1347-8613

MicroRNAs in neural stem cells and neurogenesis

岡野栄之

FRONTIERS IN NEUROSCIENCE 6 S347 - S348 2012年

研究論文（学術雑誌）, ISSN 1662-453X

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NOTCH SIGNALING AND THE DEVELOPING INNER EAR

岡野栄之

NOTCH SIGNALING IN EMBRYOLOGY AND CANCER 727 161 - 173 2012年

研究論文（学術雑誌）, ISSN 0065-2598

Usefulness of human disease-specific iPS cells research for understanding of age-related higher brain dysfunction

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 118 124P - 124P 2012年

研究論文（学術雑誌）, ISSN 1347-8613

Nestin Protein Is Phosphorylated in Adult Neural Stem/Progenitor Cells and Not Endothelial Progenitor Cells

岡野栄之

STEM CELLS INTERNATIONAL 1410 - 1422 2012年

研究論文（学術雑誌）, ISSN 1687-966X

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Renin-angiotensin system hyperactivation can induce inflammation and retinal neural dysfunction

Kurihara Toshihide, Ozawa Yoko, Ishida Susumu, Okano Hideyuki, Tsubota Kazuo

International Journal of Inflammation 2012 2012年

ISSN 2090-8040

The renin-angiotensin system (RAS) is a hormone system that has been classically known as a blood pressure regulator but is becoming well recognized as a proinflammatory mediator. In many diverse tissues, RAS pathway elements are also produced intrinsically, making it possible for tissues to respond more dynamically to systemic or local cues. While RAS is important for controlling normal inflammatory responses, hyperactivation of the pathway can cause neural dysfunction by inducing accelerated degradation of some neuronal proteins such as synaptophysin and by activating pathological glial responses. Chronic inflammation and oxidative stress are risk factors for high incidence vision-threatening diseases such as diabetic retinopathy (DR), age-related macular degeneration (AMD), and glaucoma. In fact, increasing evidence suggests that RAS inhibition may actually prevent progression of various ocular diseases including uveitis, DR, AMD, and glaucoma. Therefore, RAS inhibition may be a promising therapeutic approach to fine-tune inflammatory responses and to prevent or treat certain ocular and neurodegenerative diseases. © 2012 Toshihide Kurihara et al.

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Derivation of induced pluripotent stem cells by retroviral gene transduction in mammalian species

Imamura Masanori, Okuno Hironobu, Tomioka Ikuo, Kawamura Yoshimi, Lin Zachary Yu Ching, Nakajima Ryusuke, Akamatsu Wado, Okano Hirotaka James, Matsuzaki Yumi, Sasaki Erika, Okano Hideyuki

21 - 48 2012年

Pluripotent stem cells can provide us with an enormous cell source for in vitro model systems for development. In 2006, new methodology was designed to generate pluripotent stem cells directly from somatic cells, and these cells were named induced pluripotent stem cells (iPSCs). This method consists of technically simple procedures: donor cell preparation, gene transduction, and isolation of embryonic stem cell-like colonies. The iPSC technology enables cell biologists not only to obtain pluripotent stem cells easily but also to study the reprogramming events themselves. Here, we describe the protocols to generate iPSCs from somatic origins by using conventional viral vectors. Specifically, we state the usage of three mammalian species: mouse, common marmoset, and human. As mouse iPSC donors, fibroblasts are easily prepared, while mesenchymal stem cells are expected to give rise to highly reprogrammed iPSCs efficiently. Common marmoset (Callithrix jacchus), a nonhuman primate, represents an alternative model to the usual laboratory animals. Finally, patient-specific human iPSCs give us an opportunity to examine the pathology and mechanisms of dysregulated genomic imprinting. The iPSC technology will serve as a valuable method for studying genomic imprinting, and conversely, the insights from these studies will offer valuable criteria to assess the potential of iPSCs. © 2012 Springer Science+Business Media, LLC.

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Notch signaling and the developing inner ear

Murata Junko, Ikeda Katsuhisa, Okano Hideyuki

161 - 173 2012年

Sensory hair cells (HCs) and their associated nonsensory supporting cells (SCs) exhibit a typical mosaic pattern in each of the sensory patches in the inner ear. Notch signaling has been considered to conduct the formation of this mosaic pattern through one of its famous functions, known as 'lateral inhibition'. The two Notch ligands Delta-like1 and Jagged2 are believed to act synergistically at the stage of cell diversification in mammals. In addition, many current studies suggest that Notch signaling has another inductive, but not inhibiting, role in the determination of the prosensory region, which precedes the cell diversification of HCs and SCs and Jagged1 is thought to be an essential ligand in this process. Earlier in ear development, the first cell fate determination begins with the delamination of the neuroblasts from the otic epithelium. The delaminated neuroblasts migrate and coalesce to form cochleovestibular ganglion. Notch signaling pathway is thought to function during the delamination through its lateral inhibitory mechanism. Recently, many experiments examining Notch-related gene expression patterns and direct functional analyses of genes have revealed multiple important functions of Notch in inner ear development. Here, we survey a series of studies and discuss the issues that remain to be elucidated in the future. © 2012 Landes Bioscience and Springer Science+Business Media.

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Regeneration of the damaged CNS using human iPSCs-derived neural progenitor cells

Okano Hideyuki

Clinical Neurology 52 （ 11 ） 1147 2012年

ISSN 0009-918X

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Modeling familial alzheimer's disease with induced pluripotent stem cells

Yagi Takuya, Ito Daisuke, Okada Yohei, Akamatsu Wado, Nihei Yoshihiro, Okano Hideyuki, Suzuki Norihiro

Clinical Neurology 52 （ 11 ） 1134 - 1136 2012年

ISSN 0009-918X

Alzheimer's disease (AD) is the most common form of age-related dementia, characterized by progressive memory loss and cognitive disturbance. According to the amyloid cascade hypothesis, a prevailing theory of AD pathology, accumulation of toxic Aβ42, in the brain is the initiator of AD pathogenesis, subsequently leading to the formation of neurofibrillary tangles, and consequently neuronal loss. Mutations of presenilin 1 (PS1) and presenilin 2(PS2), which are catalytic components of γ-secretase, are causative factors for autosomal dominant early-onset familial AD (FAD). Induced pluripotent stem cell (iPSC) technology provides a new method for elucidating the molecular basis of human diseases, including neurodegenerative diseases. Here we generate iPSCs from fibroblasts of FAD patients with mutations in PS1 (A246E) and PS2 (N141I), and characterize the differentiation of these cells into neurons. We find that FAD-iPSC-derived differentiated neurons have increased toxic Aβ42 secretion, recapitulating the molecular pathogenesis of mutant presenilins. Furthermore, secretion of Aβ42 from these neurons sharply responds to γ secretase inhibitors and modulators, indicating the potential for identification and validation of candidate drugs. Our findings demonstrate that the FAD-iPSC-derived neuron is a valid model of AD and provides an innovative strategy for the study of late-onset neurodegenerative diseases.

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Investigation of pathophysiology of Parkinson's disease using iPS cells technology

Okano Hideyuki

Clinical Neurology 52 （ 11 ） 899 2012年

ISSN 0009-918X

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Cisterna-specific localization of glycosylation-related proteins to the Golgi apparatus

Yamamoto-Hino Miki, Abe Masato, Shibano Takako, Setoguchi Yuka, Awano Wakae, Ueda Ryu, Okano Hideyuki, Goto Satoshi

Cell Structure and Function 37 （ 1 ） 55 - 63 2012年

ISSN 0386-7196

The Golgi apparatus is an intracellular organelle playing central roles in post-translational modification and in the secretion of membrane and secretory proteins. These proteins are synthesized in the endoplasmic reticulum (ER) and transported to the cis-, medial- and trans-cisternae of the Golgi. While trafficking through the Golgi, proteins are sequentially modified with glycan moieties by different glycosyltransferases. Therefore, it is important to analyze the glycosylation function of the Golgi at the level of cisternae. Markers widely used for cis-, medial- and trans-cisternae/trans Golgi network (TGN) in Drosophila are GM130, 120 kDa and Syntaxin16 (Syx16); however the anti-120 kDa antibody is no longer available. In the present study, Drosophila Golgi complex-localized glycoprotein-1 (dGLG1) was identified as an antigen recognized by the anti-120 kDa antibody. A monoclonal anti-dGLG1 antibody suitable for immunohistochemistry was raised in rat. Using these markers, the localization of glycosyltransferases and nucleotide-sugar transporters (NSTs) was studied at the cisternal level. Results showed that glycosyltransferases and NSTs involved in the same sugar modification are localized to the same cisternae. Furthermore, valuable functional information was obtained on the localization of novel NSTs with as yet incompletely characterized biochemical properties, © 2012 by Japan Society for Cell Biology.

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Novel Method for Analyzing Locomotor Ability after Spinal Cord Injury in Rats: Technical Note

岡野栄之

NEUROLOGIA MEDICO-CHIRURGICA 53 （ 12 ） 907 - 913 2013年12月

研究論文（学術雑誌）, ISSN 0470-8105

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Olfactory cells via nasal biopsy reflect the developing brain in gene expression profiles: Utility and limitation of the surrogate tissues in research for brain disorders

岡野栄之

NEUROSCIENCE RESEARCH 77 （ 4 ） 247 - 250 2013年12月

研究論文（学術雑誌）, ISSN 0168-0102

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Multidimensional MRI-CT atlas of the naked mole-rat brain (Heterocephalus glaber)

岡野栄之

FRONTIERS IN NEUROANATOMY 7 2462 - 2469 2013年12月

研究論文（学術雑誌）, ISSN 1662-5129

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In vivo tracing of neural tracts in tiptoe-walking yoshimura mice by diffusion tensor tractography

Takano Morito, Komaki Yuji, Hikishima Keigo, Konomi Tsunehiko, Fujiyoshi Kanehiro, Tsuji Osahiko, Okano Hideyuki, Toyama Yoshiaki, Nakamura Masaya

107 - 117 2013年11月

In ossification of the posterior longitudinal ligaments (OPLL), axonal disruption results in motor and sensory function impairment. Twy (tiptoe-walking Yoshimura) mice develop spontaneous calcification in the cervical ligaments, thereby causing chronic compression of the spinal cords. To determine whether in vivo diffusion tensor tractography (DTT) can evaluate the axonal disruption of the chronic compressive spinal cords in twy mice, 6-, 15-, and 20-week-old twy mice were chronologically subjected to DTT. MRI was performed using a 7.0-Tesla magnet with a surface coil (CryoProbe). Diffusion tensor images were analyzed using TrackVis (Massachusetts General Hospital, MA, USA). We succeeded in depicting in vivo high-resolution DTT of the twy mice. The progress of the ligamentous calcification was observed at C2-3 level in each twy mouse, and the number of RT-97 or SMI31 positive fibers was decreased depending on the severity of the compression of the spinal cord. Quantitative analysis of sequential DTT enabled to detect subtle damage of the compressed spinal cord prior to the deterioration of neurological function in twy mice. Thus, in a clinical setting, DTT could be a new effective imaging modality in patients with cervical OPLL.

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Regenerative medicine for spinal cord injury utilizing iPS cells

Tsuji Osahiko, Nori Satoshi, Kobayashi Yoshiomi, Fujiyoshi Kanehiro, Okano Hideyuki, Toyama Yoshiaki, Nakamura Masaya

229 - 245 2013年11月

Reports of functional recovery from spinal cord injury (SCI) after the transplantation of rat-fetus-derived neural stem/precursor cells (NS/PCs) and murine embryonic stem cells (ES cells) have raised great expectations for the successful clinical trial of stem cell transplantation therapy. However, the ethical issues concerning about destroying human embryos or fertilized oocytes to obtain such stem cells have been a major impediment to developing clinically useful stem cell sources and to use them in clinical applications. Recently, induced pluripotent stem cells (iPS cells), which can serve as a source of autologous cell transplantation, have been attracting a lot of attention as a clinically practical alternative to stem cells obtained directly from tissues. In this chapter, we outline the neural induction of murine and human iPS cells, their therapeutic efficacy in mouse and primate SCI models, and their safety in vivo.

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Recombinant human hepatocyte growth factor promotes functional recovery after spinal cord injury

Kitamura Kazuya, Iwanami Akio, Fujiyoshi Kanehiro, Yamane Jun Ichi, Hikishima Keigo, Funakoshi Hiroshi, Nakamura Toshikazu, Aoki Masashi, Toyama Yoshiaki, Okano Hideyuki, Nakamura Masaya

147 - 167 2013年11月

Hepatocyte growth factor (HGF), which is a potent mitogen for mature hepatocytes and a mediator of the inflammatory responses to tissue injury, was recently highlighted as a potent neurotrophic factor in the central nervous system. We revealed that introducing exogenous HGF into the injured rat spinal cord using a herpes simplex virus-1 vector significantly reduces the area of damaged tissue and promotes functional recovery. However, this rat study did not examine the therapeutic effects of administering HGF after injury, which is the most critical issue for clinical application. To translate this strategy to human treatment, we induced a contusive cervical SCI in the common marmoset, a primate, and then administered recombinant human HGF (rhHGF) intrathecally. Motor function was assessed using an original open-field scoring system focusing on manual function, including reach-and-grasp performance and hand placement in walking. The intrathecal rhHGF preserved the corticospinal fibers and myelinated areas, thereby promoting functional recovery. This study demonstrates the therapeutic effects, safety, and clinical efficacy of intrathecal rhHGF treatment for SCI in adult nonhuman primates and the possibility that this novel therapy may be suitable for clinical application.

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Assessment of injured spinal cord using diffusion tensor tractography

Fujiyoshi Kanehiro, Konomi Tsunehiko, Tsuji Osahiko, Yamada Masayuki, Hikishima Keigo, Momoshima Suketaka, Okano Hideyuki, Toyama Yoshiaki, Nakamura Masaya

345 - 365 2013年11月

In spinal cord injury (SCI), the evaluation of axonal fibers is important to assess the severity of injury and efficacy of any treatment protocol, but conventional methods such as tracer injection in the brain parenchyma are highly invasive and require histological evaluation, precluding clinical applications. Magnetic resonance imaging (MRI) is essential for predicting prognosis and planning treatment of patients with SCI noninvasively. However, the information provided by conventional T1- and T2-weighted MRI of the spinal cord is essentially limited to the differentiation of the white matter from the gray matter. By contrast, diffusion-weighted magnetic resonance imaging (DWI) provides much information about biological structures. In particular, diffusion tensor imaging (DTI) and diffusion tensor tractography (DTT) are powerful tools for evaluating white matter fibers in the central nervous system. We previously established a reproducible SCI model in adult common marmosets and demonstrated that DTT could be used to trace the neural tracts in the intact and injured spinal cord of these animals in vivo. Recently, many reports using DTT to analyze the spinal cord area have been published. Based on the findings from our experimental studies, we are now routinely performing DTT of the human spinal cord clinically. In this chapter, we outline the basic principles of DTT and describe the characteristics, limitations, and clinical application of DTT in the spinal cord.

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Chondroitin sulphate N-acetylgalactosaminyl-transferase-1 inhibits recovery from neural injury

岡野栄之

NATURE COMMUNICATIONS 4 106 - 114 2013年11月

研究論文（学術雑誌）, ISSN 2041-1723

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A homozygous nonsense mutation in the gene for Tmem79, a component for the lamellar granule secretory system, produces spontaneous eczema in an experimental model of atopic dermatitis

岡野栄之

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY 132 （ 5 ） 1111 - + 2013年11月

研究論文（学術雑誌）, ISSN 0091-6749

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Role of HIF-1 alpha-activated Epac1 on HSC-mediated neuroplasticity in stroke model

岡野栄之

NEUROBIOLOGY OF DISEASE 58 76 - 91 2013年10月

研究論文（学術雑誌）, ISSN 0969-9961

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Differential cellular localization of antioxidant enzymes in the trigeminal ganglion

Sato H., Shibata M., Shimizu T., Shibata S., Toriumi H., Ebine T., Kuroi T., Iwashita T., Funakubo M., Kayama Y., Akazawa C., Wajima K., Nakagawa T., Okano H., Suzuki N.

Neuroscience 248 345 - 358 2013年09月

ISSN 0306-4522

Because of its high oxygen demands, neural tissue is predisposed to oxidative stress. Here, our aim was to clarify the cellular localization of antioxidant enzymes in the trigeminal ganglion. We found that the transcriptional factor Sox10 is localized exclusively in satellite glial cells (SGCs) in the adult trigeminal ganglion. The use of transgenic mice that express the fluorescent protein Venus under the Sox10 promoter enabled us to distinguish between neurons and SGCs. Although both superoxide dismutases 1 and 2 were present in the neurons, only superoxide dismutase 1 was identified in SGCs. The enzymes relevant to hydrogen peroxide degradation displayed differential cellular localization, such that neurons were endowed with glutathione peroxidase 1 and thioredoxin 2, and catalase and thioredoxin 2 were present in SGCs. Our immunohistochemical finding showed that only SGCs were labeled by the oxidative damage marker 8-hydroxy-2'-deoxyguanosine, which indicates that the antioxidant systems of SGCs were less potent. The transient receptor potential vanilloid subfamily member 1 (TRPV1), the capsaicin receptor, is implicated in inflammatory hyperalgesia, and we demonstrated that topical capsaicin application causes short-lasting mechanical hyperalgesia in the face. Our cell-based assay revealed that TRPV1 agonist stimulation in the presence of TRPV1 overexpression caused reactive oxygen species-mediated caspase-3 activation. Moreover, capsaicin induced the cellular demise of primary TRPV1-positive trigeminal ganglion neurons in a dose-dependent manner, and this effect was inhibited by a free radical scavenger and a pancaspase inhibitor. This study delineates the localization of antioxidative stress-related enzymes in the trigeminal ganglion and reveals the importance of the pivotal role of reactive oxygen species in the TRPV1-mediated caspase-dependent cell death of trigeminal ganglion neurons. Therapeutic measures for antioxidative stress should be taken to prevent damage to trigeminal primary sensory neurons in inflammatory pain disorders. © 2013 IBRO.

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[110th Scientific Meeting of the Japanese Society of Internal Medicine

Nakamura Masaya, Okano Hideyuki, Toyama Yoshiaki

Nihon Naika Gakkai zasshi. The Journal of the Japanese Society of Internal Medicine 102 （ 9 ） 2247 - 2253 2013年09月

ISSN 0021-5384

The effect of SCN1A mutations on patient-derived GABAergic neurons

Higurashi Norimichi, Okano Hideyuki, Hirose Shinichi

Future Neurology 8 （ 5 ） 487 - 489 2013年09月

ISSN 1479-6708

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Sox6 Up-Regulation by Macrophage Migration Inhibitory Factor Promotes Survival and Maintenance of Mouse Neural Stem/Progenitor Cells

岡野栄之

PLOS ONE 8 （ 9 ） 279 - 284 2013年09月

研究論文（学術雑誌）, ISSN 1932-6203

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Sema3A regulates bone-mass accrual through sensory innervations (vol 497, pg 490, 2013)

岡野栄之

NATURE 500 （ 7464 ） 855 - 864 2013年08月

研究論文（学術雑誌）, ISSN 0028-0836

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SOX10 is a novel marker of acinus and intercalated duct differentiation in salivary gland tumors: a clue to the histogenesis for tumor diagnosis

岡野栄之

MODERN PATHOLOGY 26 （ 8 ） 1041 - 1050 2013年08月

研究論文（学術雑誌）, ISSN 0893-3952

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LNGFR(+)THY-1(+)VCAM-1(hi+) Cells Reveal Functionally Distinct Subpopulations in Mesenchymal Stem Cells

岡野栄之

STEM CELL REPORTS 1 （ 2 ） 152 - 165 2013年08月

研究論文（学術雑誌）, ISSN 2213-6711

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The long non-coding RNA nuclear-enriched abundant transcript 1_2 induces paraspeckle formation in the motor neuron during the early phase of amyotrophic lateral sclerosis

岡野栄之

MOLECULAR BRAIN 6 13064 - 13069 2013年07月

研究論文（学術雑誌）, ISSN 1756-6606

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Chromatin Regulator PRC2 Is a Key Regulator of Epigenetic Plasticity in Glioblastoma

岡野栄之

CANCER RESEARCH 73 （ 14 ） 4559 - 4570 2013年07月

研究論文（学術雑誌）, ISSN 0008-5472

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Development and Characterization of cDNA Resources for the Common Marmoset: One of the Experimental Primate Models

岡野栄之

DNA RESEARCH 20 （ 3 ） 255 - 262 2013年06月

研究論文（学術雑誌）, ISSN 1340-2838

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Kidney Specific Protein-Positive Cells Derived from Embryonic Stem Cells Reproduce Tubular Structures In Vitro and Differentiate into Renal Tubular Cells

岡野栄之

PLOS ONE 8 （ 6 ） 811 - 821 2013年06月

研究論文（学術雑誌）, ISSN 1932-6203

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Human Induced Pluripotent Stem Cell-Derived Ectodermal Precursor Cells Contribute to Hair Follicle Morphogenesis In Vivo

岡野栄之

JOURNAL OF INVESTIGATIVE DERMATOLOGY 133 （ 6 ） 1479 - 1488 2013年06月

研究論文（学術雑誌）, ISSN 0022-202X

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Musashi1 as a potential therapeutic target and diagnostic marker for lung cancer

岡野栄之

ONCOTARGET 4 （ 5 ） 739 - 750 2013年05月

研究論文（学術雑誌）, ISSN 1949-2553

The Drosophila Ortholog of MLL3 and MLL4, trithorax related, Functions as a Negative Regulator of Tissue Growth

岡野栄之

MOLECULAR AND CELLULAR BIOLOGY 33 （ 9 ） 1702 - 1710 2013年05月

研究論文（学術雑誌）, ISSN 0270-7306

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A human Dravet syndrome model from patient induced pluripotent stem cells

岡野栄之

MOLECULAR BRAIN 6 1312 - 1313 2013年05月

研究論文（学術雑誌）, ISSN 1756-6606

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Correction

Muto Jun, Imai Takao, Ogawa Daisuke, Nishimoto Yoshinori, Okada Yohei, Mabuchi Yo, Kawase Takeshi, Iwanami Akio, Mischel Paul S., Saya Hideyuki, Yoshida Kazunari, Matsuzaki Yumi, Okano Hideyuki

PLoS ONE 8 （ 4 ） 2013年04月

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Diffusion tensor imaging and tractography of the spinal cord: From experimental studies to clinical application

岡野栄之

EXPERIMENTAL NEUROLOGY 242 74 - 82 2013年04月

研究論文（学術雑誌）, ISSN 0014-4886

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Modulation of the shape and speed of a chemical wave in an unstirred Belousov-Zhabotinsky reaction by a rotating magnet

岡野栄之

BIOELECTROMAGNETICS 34 （ 3 ） 220 - 230 2013年04月

研究論文（学術雑誌）, ISSN 0197-8462

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Notch Inhibition Induces Cochlear Hair Cell Regeneration and Recovery of Hearing after Acoustic Trauma (vol 77, pg 58, 2013)

岡野栄之

NEURON 78 （ 2 ） 403 - 403 2013年04月

研究論文（学術雑誌）, ISSN 0896-6273

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PML mediates glioblastoma resistance to mammalian target of rapamycin (mTOR)-targeted therapies

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 110 （ 11 ） 4339 - 4344 2013年03月

研究論文（学術雑誌）, ISSN 0027-8424

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PML mediates glioblastoma resistance to mammalian target of rapamycin (mTOR)-targeted therapies

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 110 （ 11 ） 4339 - 4344 2013年03月

研究論文（学術雑誌）, ISSN 0027-8424

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Aberrant Gene Expression and Sexually Incompatible Genomic Imprinting in Oocytes Derived from XY Mouse Embryonic Stem Cells In Vitro

岡野栄之

PLOS ONE 8 （ 3 ） 116 - 125 2013年03月

研究論文（学術雑誌）, ISSN 1932-6203

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Functional Corneal Endothelium Derived from Corneal Stroma Stem Cells of Neural Crest Origin by Retinoic Acid and Wnt/beta-Catenin Signaling

岡野栄之

STEM CELLS AND DEVELOPMENT 22 （ 5 ） 682 - 696 2013年03月

研究論文（学術雑誌）, ISSN 1547-3287

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Enhanced Aggregation of Androgen Receptor in Induced Pluripotent Stem Cell-derived Neurons from Spinal and Bulbar Muscular Atrophy

岡野栄之

JOURNAL OF BIOLOGICAL CHEMISTRY 288 （ 12 ） 8043 - 8052 2013年03月

研究論文（学術雑誌）, ISSN 0021-9258

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Assessment of homozygosity levels in the mito-gynogenetic torafugu (Takifugu rubripes) by genome-wide SNP analyses

岡野栄之

AQUACULTURE 380 114 - 119 2013年03月

研究論文（学術雑誌）, ISSN 0044-8486

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Epigenetic transcriptional activation of monocyte chemotactic protein 3 contributes to long-lasting neuropathic pain

岡野栄之

BRAIN 136 828 - 843 2013年03月

研究論文（学術雑誌）, ISSN 0006-8950

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[Reconstruction for spinal cord injury].

Hori Keiko, Nakamura Masaya, Okano Hideyuki

Nihon Jibiinkoka Gakkai kaiho 116 （ 2 ） 53 - 59 2013年02月

ISSN 0030-6622

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Semaphorin 3A regulates bone homeostasis through sensory nerve system

岡野栄之

JOURNAL OF BONE AND MINERAL RESEARCH 28 S154 - S154 2013年02月

研究論文（学術雑誌）, ISSN 0884-0431

Steps Toward Safe Cell Therapy Using Induced Pluripotent Stem Cells

岡野栄之

CIRCULATION RESEARCH 112 （ 3 ） 523 - 533 2013年02月

研究論文（学術雑誌）, ISSN 0009-7330

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Atlas of the developing brain of the marmoset monkey constructed using magnetic resonance histology

Hikishima K., Sawada K., Murayama A. Y., Komaki Y., Kawai K., Sato N., Inoue T., Itoh T., Momoshima S., Iriki A., Okano H. J., Sasaki E., Okano H.

Neuroscience 230 102 - 113 2013年01月

ISSN 0306-4522

The developmental anatomy of the brain is largely directed by neural-based cues. Despite this knowledge, the developmental trajectory of the primate brain has not yet been fully characterized. To realize this goal, the advance in noninvasive imaging methods and new brain atlases are essential. The common marmoset (Callithrix jacchus), a small New World primate, is widely used in neuroscience research. The recent introduction of transgenic techniques has enabled the marmoset to be used as a genetically modifiable primate model for brain development. Here, a magnetic resonance histology technique involving the use of ultra-high-resolution ex vivo magnetic resonance imaging (MRI) was performed to identify the developmental anatomy of the marmoset brain at different time points from gestational week 8 through to birth. The data allowed the generation of a multidimensional atlas of brain structures at different developmental stages. Furthermore, in utero MRI techniques were developed to noninvasively monitor brain development during the embryonic and fetal stages. The multidimensional atlas and the MRI tools developed herein are anticipated to further our understanding of the developing primate brain. © 2012 IBRO.

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Cell-intrinsic reprogramming capability

Imamura Masanori, Lin Zachary Yu Ching, Okano Hideyuki

Reproductive Medicine and Biology 12 （ 1 ） 1 - 14 2013年01月

ISSN 1445-5781

In multicellular organisms, germ cells are an extremely specialized cell type with the vital function of transmitting genetic information across generations. In this respect, they are responsible for the perpetuity of species, and are separated from somatic lineages at each generation. Interestingly, in the past two decades research has shown that germ cells have the potential to proceed along two distinct pathways: gametogenesis or pluripotency. Unequivocally, the primary role of germ cells is to produce gametes, the sperm or oocyte, to produce offspring. However, under specific conditions germ cells can become pluripotent, as shown by teratoma formation in vivo or cell culture-induced reprogramming in vitro. This phenomenon seems to be a general propensity of germ cells, irrespective of developmental phase. Recent attempts at cellular reprogramming have resulted in the generation of induced pluripotent stem cells (iPSCs). In iPSCs, the intracellular molecular networks instructing pluripotency have been activated and override the exclusively somatic cell programs that existed. Because the generation of iPSCs is highly artificial and depends on gene transduction, whether the resulting machinery reflects any physiological cell-intrinsic programs is open to question. In contrast, germ cells can spontaneously shift their fate to pluripotency during in-vitro culture. Here, we review the two fates of germ cells, i.e., differentiation and reprogramming. Understanding the molecular mechanisms regulating differentiation versus reprogramming would provide invaluable insight into understanding the mechanisms of cellular reprogramming that generate iPSCs. © 2012 Japan Society for Reproductive Medicine.

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Musashi-1 Post-Transcriptionally Enhances Phosphotyrosine-Binding Domain-Containing m-Numb Protein Expression in Regenerating Gastric Mucosa

岡野栄之

PLOS ONE 8 （ 1 ） 1669 - 1681 2013年01月

研究論文（学術雑誌）, ISSN 1932-6203

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Time-dependent changes in the microenvironment of injured spinal cord affects the therapeutic potential of neural stem cell transplantation for spinal cord injury

岡野栄之

MOLECULAR BRAIN 6 1451 - 1464 2013年01月

研究論文（学術雑誌）, ISSN 1756-6606

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Dose-dependent roles for canonical Wnt signalling in de novo crypt formation and cell cycle properties of the colonic epithelium

岡野栄之

DEVELOPMENT 140 （ 1 ） 66 - 75 2013年01月

研究論文（学術雑誌）, ISSN 0950-1991

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In Vivo Tracing of Neural Tracts in Tiptoe Walking Yoshimura Mice by Diffusion Tensor Tractography

岡野栄之

SPINE 38 （ 2 ） E66 - E72 2013年01月

研究論文（学術雑誌）, ISSN 0362-2436

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Cell transplantation therapies for spinal cord injury focusing on induced pluripotent stem cells

岡野栄之

CELL RESEARCH 23 （ 1 ） 70 - 80 2013年01月

研究論文（学術雑誌）, ISSN 1001-0602

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Human induced pluripotent stem (iPS) cells-derived neural stem cell bundle covered with growth factor-encapsulated amphiphilic chitosan

Kato-Negishi Midori, Onoe Hiroaki, Iwanaga Shintaroh, Kobayashi Yoshiomi, Nakamura Masaya, Okano Hideyuki, Takeuchi Shoji

1054 - 1056 2013年

This paper describes human induced pluripotent stem (iPS) cell-derived neural stem cell (hiPSC-NSC) bundles covered with growth factor-encapsulated amphiphilic chitosan. The hiPSC-NSC bundle is formed by 4-12 hiPSC-NSC microfibers that are made by core-shell hydrogel microfibers encapsulating hiPSC-NSCs. The hiPSC-NSC microfibers can be cultured for over two weeks and induced to differentiate into neurons predominantly. We modified the coating material to make bundle structures that have growth factor enrichment capability (Figure 1). The coating material, amphiphilic chitosan, can release growth factors slowly for a long time, allowing hiPSC-NSCs to survive, grow and differentiate into neural lineages in the bundle structure. We believe that our hiPSC-NSC bundle would be an extremely effective tool for neural transplantation in neurodegenerative diseases such as stroke, spinal cord injury and peripheral nerve injury, because hiPSC-NSCs in the bundle structure have long-term survival ability in vivo.

Identification and analysis of target genes of the Rett syndrome causative gene, Mecp2, in the cerebral cortex

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 63 S25 - S25 2013年

研究論文（学術雑誌）, ISSN 1880-6546

Multiple profiling of changes in miRNA expression under the neuropathic pain

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 121 121P - 121P 2013年

研究論文（学術雑誌）, ISSN 1347-8613

Static Magnetic Field Effects on Impaired Peripheral Vasomotion in Conscious Rats

岡野栄之

EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE S54 - S54 2013年

研究論文（学術雑誌）, ISSN 1741-427X

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Purinergic Regulation of Astrocytic Phagocytosis

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 121 74P - 74P 2013年

研究論文（学術雑誌）, ISSN 1347-8613

Conditional deletion of STAT3 in astrocytes suppresses reactive astrocytes after nerve injury and neuropathic pain

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 121 60P - 60P 2013年

研究論文（学術雑誌）, ISSN 1347-8613

Epigenetic transcriptional activation of chemokines in astrocytes is critical for neuropathic pain

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 121 109P - 109P 2013年

研究論文（学術雑誌）, ISSN 1347-8613

Role of dopamine receptors in ES and iPS cell differentiation

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 121 76P - 76P 2013年

研究論文（学術雑誌）, ISSN 1347-8613

iPS technology-based cell therapy for damaged CNS and investigation of neural disorders

岡野栄之

JOURNAL OF PHYSIOLOGICAL SCIENCES 63 S85 - S85 2013年

研究論文（学術雑誌）, ISSN 1880-6546

Prospective Isolation of Murine and Human Bone Marrow Mesenchymal Stem Cells Based on Surface Markers

岡野栄之

STEM CELLS INTERNATIONAL S269 - S269 2013年

研究論文（学術雑誌）, ISSN 1687-966X

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Current status and perspective on regenerative medicine for spinal cord injury using iPS cell

Nakamura Masaya, Toyama Yoshiaki, Okano Hideyuki

Clinical Neurology 53 （ 11 ） 1013 - 1016 2013年

ISSN 0009-918X

Stimulated by the 2012 Nobel Prize in Physiology or Medicine awarded for Shinya Yamanaka and Sir John Gurdon, there is an increasing interest in the iPS cells and reprogramming technologies in medical science. While iPS cells are expected to open new era providing enormous opportunities in the biomedical sciences in terms of cell therapies for regenerative medicine, safety-related concerns for iPS cell-based cell therapy should be resolved prior to the clinical application of iPS cells. In this symposium, the pre-clinical investigations of cell therapy for SCI using neural stem/progenitor cells derived from iPS cells, and their safety issues in vivo are outlined.

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Generation of human melanocytes from induced pluripotent stem cells

Ohta Shigeki, Imaizumi Yoichi, Akamatsu Wado, Okano Hideyuki, Kawakami Yutaka

193 - 215 2013年

The discovery of human induced pluripotent stem cells (iPSCs) has provided a model system for studying early events during human development. Developmentally melanocytes originate from migratory neural crest cells that emerge from the neural plate during embryogenesis after a complex process of differentiation, proliferation, and migration out of the neural tube along defined pathways. In the adult, human melanocytes are located in the basal layer of the epidermis, hair follicles, uvea, inner ear, and meninges. In the epidermis, melanocytes produce melanin pigment that gives color to the skin as well as providing protection from ultraviolet light damage. In addition, melanocytes transfer melanin pigment to hair matrix keratinocytes during each hair cycle to maintain hair pigmentation. Characterization of mouse melanocyte stem cells (MELSCs) is more complete than for humans. MELSCs are located in the bulge region of hair follicles, where hair follicle stem cells (HFSCs) also reside. Recently, it has been demonstrated that HFSCs provide a functional nice for MELSCs. According to current cancer stem cell theory, melanomas are considered to evolve from MELSCs, although the exact mechanism remains to be elucidated fully. In humans, importantly, the lack of more specific markers of MELSCs, current understanding of the molecular regulations of melanocyte development remains incomplete. Recently, the generation of melanocytes from iPSCs has lead to some clarification of human melanocyte development in vitro. Utilization of iPSC-derived melanocytes may prove invaluable in further study of human melanocytic development and novel therapies for patients suffering with pigmentation disorders and melanoma. © 2013 Springer Science+Business Media New York.

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Impaired hematopoietic differentiation of RUNX1-mutated induced pluripotent stem cells derived from FPD/AML patients

Sakurai M., Kunimoto H., Watanabe N., Fukuchi Y., Yuasa S., Yamazaki S., Nishimura T., Sadahira K., Fukuda K., Okano H., Nakauchi H., Morita Y., Matsumura I., Kudo K., Ito E., Ebihara Y., Tsuji K., Harada Y., Harada H., Okamoto S., Nakajima H.

Leukemia 28 （ 12 ） 2344 - 2354 2014年12月

ISSN 0887-6924

Somatic mutation of RUNX1 is implicated in various hematological malignancies, including myelodysplastic syndrome and acute myeloid leukemia (AML), and previous studies using mouse models disclosed its critical roles in hematopoiesis. However, the role of RUNX1 in human hematopoiesis has never been tested in experimental settings. Familial platelet disorder (FPD)/AML is an autosomal dominant disorder caused by germline mutation of RUNX1, marked by thrombocytopenia and propensity to acute leukemia. To investigate the physiological function of RUNX1 in human hematopoiesis and pathophysiology of FPD/AML, we derived induced pluripotent stem cells (iPSCs) from three distinct FPD/AML pedigrees (FPD-iPSCs) and examined their defects in hematopoietic differentiation. By in vitro differentiation assays, FPD-iPSCs were clearly defective in the emergence of hematopoietic progenitors and differentiation of megakaryocytes, and overexpression of wild-type (WT)-RUNX1 reversed most of these phenotypes. We further demonstrated that overexpression of mutant-RUNX1 in WT-iPSCs did not recapitulate the phenotype of FPD-iPSCs, showing that the mutations were of loss-of-function type. Taken together, this study demonstrated that haploinsufficient RUNX1 allele imposed cell-intrinsic defects on hematopoietic differentiation in human experimental settings and revealed differential impacts of RUNX1 dosage on human and murine megakaryopoiesis. FPD-iPSCs will be a useful tool to investigate mutant RUNX1-mediated molecular processes in hematopoiesis and leukemogenesis.

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Inflammatory and immune responses in the cochlea: potential therapeutic targets for sensorineural hearing loss

岡野栄之

FRONTIERS IN PHARMACOLOGY 5 70 - 80 2014年12月

研究論文（学術雑誌）, ISSN 1663-9812

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Three-dimensional motion analysis of arm-reaching movements in healthy and hemispinalized common marmosets

岡野栄之

BEHAVIOURAL BRAIN RESEARCH 275 259 - 268 2014年12月

研究論文（学術雑誌）, ISSN 0166-4328

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Nuclear receptor gene alteration in human induced pluripotent stem cells with hepatic differentiation propensity

岡野栄之

HEPATOLOGY RESEARCH 44 （ 14 ） E408 - E419 2014年12月

研究論文（学術雑誌）, ISSN 1386-6346

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Characterization of transposable element regulating small RNAs in the adult testes of the common marmoset

岡野栄之

GENES & GENETIC SYSTEMS 89 （ 6 ） 286 - 286 2014年12月

研究論文（学術雑誌）, ISSN 1341-7568

The Potential of Using Induced Pluripotent Stem Cells in Skin Diseases

Ohta Shigeki, Veraitch Ophelia, Okano Hideyuki, Ohyama Manabu, Kawakami Yutaka

223 - 245 2014年11月

The seminal discovery of induced pluripotent stem cells (iPSCs) and subsequent expansion in iPSC research have greatly aided our understanding of stem cell biology and have catapulted the changing face of regenerative medicine into prominence. With easy accessibility and visibility, the skin has been considered a favorable organ to study stem cell behaviors. In particular, the skin provides valuable materials for tissue engineering and testing of novel therapeutic approaches including gene therapy and regenerative medicine. The skin consists of multiple cell lineages, including interfollicular/follicular keratinocytes, melanocytes, sebocytes, and follicular/intradermal mesenchymal cells. Nowadays, iPSCs can be generated from various skin-derived cell populations. Elegant label-tracking experiments have successfully identified their stem/early precursor cell populations. Following molecular and cellular biological experimentations and transgenic studies the elucidation of necessary factors, signaling networks, environments supporting their behavior, and interactions in organogenesis and homeostasis have been uncovered. Taking advantage of the knowledge accumulated by these works, iPSC-derived keratinocytes and melanocytes have been successfully induced, which holds promise as future materials for the bioengineering of the skin and hair, and investigation of biology of skin stem cells. Recently, the concept of cancer stem cells (CSCs) has been proposed. Cancer stem cells have been implied to originate from stem cell subsets and, accordingly, iPSCs may be converted to skin CSCs, especially most life-threatening melanoma CSCs. Those iPSC-derived CSCs may significantly enhance our understanding of cancer biology and allow the development of novel therapeutic strategies. Recently, patient-specific iPSCs were established from intractable genodermatoses, including epidermolysis bullosa, and corrected for their genetic defects. Thus, iPSC technology should open a new era of stem cell biology, genetic/regenerative medicine and oncology in the field of dermatology.

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Global gene expression analysis following spinal cord injury in non-human primates

岡野栄之

EXPERIMENTAL NEUROLOGY 261 171 - 179 2014年11月

研究論文（学術雑誌）, ISSN 0014-4886

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Cadherin-7 Regulates Mossy Fiber Connectivity in the Cerebellum

岡野栄之

CELL REPORTS 9 （ 1 ） 311 - 323 2014年10月

研究論文（学術雑誌）, ISSN 2211-1247

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Spatiotemporal Expression of TRPM4 in the Mouse Cochlea

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 92 （ 10 ） 1409 - 1418 2014年10月

研究論文（学術雑誌）, ISSN 0360-4012

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Production of an immunodeficient common marmoset by ZFN-based mutagenesis

岡野栄之

TRANSGENIC RESEARCH 23 （ 5 ） 873 - 874 2014年10月

研究論文（学術雑誌）, ISSN 0962-8819

Regeneration of the damaged central nervous system through reprogramming technology: Basic concepts and potential application for cell replacement therapy

岡野栄之

EXPERIMENTAL NEUROLOGY 260 12 - 18 2014年10月

研究論文（学術雑誌）, ISSN 0014-4886

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Dramatic improvement in genome assembly achieved using doubled-haploid genomes

岡野栄之

SCIENTIFIC REPORTS 4 36 - 44 2014年10月

研究論文（学術雑誌）, ISSN 2045-2322

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Stem Cell Research and Regenerative Medicine in 2014: First Year of Regenerative Medicine in Japan

岡野栄之

STEM CELLS AND DEVELOPMENT 23 （ 18 ） 2127 - 2128 2014年09月

研究論文（学術雑誌）, ISSN 1547-3287

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Diffusion-Tensor MRI and Tractography of the Esophageal Wall Ex Vivo

岡野栄之

JOURNAL OF MAGNETIC RESONANCE IMAGING 40 （ 3 ） 567 - 576 2014年09月

研究論文（学術雑誌）, ISSN 1053-1807

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Neural Stem Cell Transplantation Improved Delayed Gastric Emptying in Apolipoprotein E-Knockout Mice

岡野栄之

DIGESTIVE DISEASES AND SCIENCES 59 （ 9 ） 2019 - 2019 2014年09月

研究論文（学術雑誌）, ISSN 0163-2116

Regulatory Factor X Transcription Factors Control Musashi1 Transcription in Mouse Neural Stem/Progenitor Cells

岡野栄之

STEM CELLS AND DEVELOPMENT 23 （ 18 ） 2250 - 2261 2014年09月

研究論文（学術雑誌）, ISSN 1547-3287

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Focal Transplantation of Human iPSC-Derived Glial-Rich Neural Progenitors Improves Lifespan of ALS Mice

岡野栄之

STEM CELL REPORTS 3 （ 2 ） 242 - 249 2014年08月

研究論文（学術雑誌）, ISSN 2213-6711

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Leptin Receptor Makes Its Mark on MSCs

岡野栄之

CELL STEM CELL 15 （ 2 ） 112 - 114 2014年08月

研究論文（学術雑誌）, ISSN 1934-5909

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Small RNA profiling and characterization of piRNA clusters in the adult testes of the common marmoset, a model primate

岡野栄之

RNA 20 （ 8 ） 1223 - 1237 2014年08月

研究論文（学術雑誌）, ISSN 1355-8382

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Esophageal Carcinoma: Ex Vivo Evaluation with Diffusion-Tensor MR Imaging and Tractography at 7 T

岡野栄之

RADIOLOGY 272 （ 1 ） 164 - 173 2014年07月

研究論文（学術雑誌）, ISSN 0033-8419

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Yb Integrates piRNA Intermediates and Processing Factors into Perinuclear Bodies to Enhance piRISC Assembly

岡野栄之

CELL REPORTS 8 （ 1 ） 103 - 113 2014年07月

研究論文（学術雑誌）, ISSN 2211-1247

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Thoracic and thoracoabdominal aortic repair under regional spinal cord hypothermia

岡野栄之

EUROPEAN JOURNAL OF CARDIO-THORACIC SURGERY 46 （ 1 ） 40 - 43 2014年07月

研究論文（学術雑誌）, ISSN 1010-7940

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Human speech- and reading-related genes display partially overlapping expression patterns in the marmoset brain

岡野栄之

BRAIN AND LANGUAGE 133 26 - 38 2014年06月

研究論文（学術雑誌）, ISSN 0093-934X

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Proposing a new RNA quadruplex structure: j-motif, with possible links to neural development

岡野栄之

JOURNAL OF BIOCHEMISTRY 155 （ 6 ） 385 - 392 2014年06月

研究論文（学術雑誌）, ISSN 0021-924X

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Astrocytic Activation in the Anterior Cingulate Cortex is Critical for Sleep Disorder Under Neuropathic Pain

岡野栄之

SYNAPSE 68 （ 6 ） 235 - 247 2014年06月

研究論文（学術雑誌）, ISSN 0887-4476

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The use of induced pluripotent stem cells to reveal pathogenic gene mutations and explore treatments for retinitis pigmentosa

岡野栄之

MOLECULAR BRAIN 7 1312 - 1313 2014年06月

研究論文（学術雑誌）, ISSN 1756-6606

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Involvement of ER Stress in Dysmyelination of Pelizaeus-Merzbacher Disease with PLP1 Missense Mutations Shown by iPSC-Derived Oligodendrocytes

岡野栄之

STEM CELL REPORTS 2 （ 5 ） 648 - 661 2014年05月

研究論文（学術雑誌）, ISSN 2213-6711

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Modeling human neurological disorders with induced pluripotent stem cells

岡野栄之

JOURNAL OF NEUROCHEMISTRY 129 （ 3 ） 388 - 399 2014年05月

研究論文（学術雑誌）, ISSN 0022-3042

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Differentiation, polarization, and migration of human induced pluripotent stem cell-derived neural progenitor cells co-cultured with a human glial cell line with radial glial-like characteristics

岡野栄之

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 447 （ 4 ） 683 - 688 2014年05月

研究論文（学術雑誌）, ISSN 0006-291X

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Regulation of PSCA expression by m-Numb in the stomach

岡野栄之

FASEB JOURNAL 28 （ 1 ） 442 - 442 2014年04月

研究論文（学術雑誌）, ISSN 0892-6638

Characterization of common marmoset dysgerminoma-like tumor induced by the lentiviral expression of reprogramming factors

岡野栄之

CANCER SCIENCE 105 （ 4 ） 402 - 408 2014年04月

研究論文（学術雑誌）, ISSN 1349-7006

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Reprogramming non-human primate somatic cells into functional neuronal cells by defined factors

岡野栄之

MOLECULAR BRAIN 7 257 - 267 2014年04月

研究論文（学術雑誌）, ISSN 1756-6606

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Birth of Healthy Offspring following ICSI in In Vitro-Matured Common Marmoset (Callithrix jacchus) Oocytes

岡野栄之

PLOS ONE 9 （ 4 ） 3674 - 3686 2014年04月

研究論文（学術雑誌）, ISSN 1932-6203

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Promise of Human Induced Pluripotent Stem Cells in Skin Regeneration and Investigation

岡野栄之

JOURNAL OF INVESTIGATIVE DERMATOLOGY 134 （ 3 ） 605 - 609 2014年03月

研究論文（学術雑誌）, ISSN 0022-202X

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Inflammatory cascades mediate synapse elimination in spinal cord compression

岡野栄之

JOURNAL OF NEUROINFLAMMATION 11 63 - 73 2014年03月

研究論文（学術雑誌）, ISSN 1742-2094

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iPS cell technologies: significance and applications to CNS regeneration and disease

岡野栄之

MOLECULAR BRAIN 7 106 - 114 2014年03月

研究論文（学術雑誌）, ISSN 1756-6606

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Prenatal Deletion of the RNA-Binding Protein HuD Disrupts Postnatal Cortical Circuit Maturation and Behavior

岡野栄之

JOURNAL OF NEUROSCIENCE 34 （ 10 ） 3674 - 3686 2014年03月

研究論文（学術雑誌）, ISSN 0270-6474

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Rewiring of regenerated axons by combining treadmill training with semaphorin3A inhibition

岡野栄之

MOLECULAR BRAIN 7 8043 - 8052 2014年03月

研究論文（学術雑誌）, ISSN 1756-6606

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Pharmacological Inhibition of Cochlear Mitochondrial Respiratory Chain Induces Secondary Inflammation in the Lateral Wall: A Potential Therapeutic Target for Sensorineural Hearing Loss

岡野栄之

PLOS ONE 9 （ 3 ） 163 - 175 2014年03月

研究論文（学術雑誌）, ISSN 1932-6203

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Evolutionarily Dynamic Alternative Splicing of GPR56 Regulates Regional Cerebral Cortical Patterning

岡野栄之

SCIENCE 343 （ 6172 ） 764 - 768 2014年02月

研究論文（学術雑誌）, ISSN 0036-8075

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Fetal sulcation and gyrification in common marmosets (Callithrix jacchus) obtained by ex vivo magnetic resonance imaging

Sawada K., Hikishima K., Murayama A. Y., Okano H. J., Sasaki E., Okano H.

Neuroscience 257 158 - 174 2014年01月

ISSN 0306-4522

The present study characterized fetal sulcation patterns and gyrification in the cerebrum of the New World monkey group, common marmosets, using a 3D T₂-weighted high-resolution anatomical magnetic resonance imaging (MRI) sequence from the fixed brain at 7-tesla ex vivo. Fetal sulcation in the marmoset cerebrum began to indent the lateral fissure and hippocampal sulcus in gestational week (GW) 12, and then the following sulci emerged: the callosal and calcarine sulci on GW 15; the superior temporal sulcus on GW 17; and the circular and occipitotemporal sulci on GW 18. The degree of cortical convolution was evaluated quantitatively based on 2D MRI slices by the gyrification index (GI) and based on 3D MRI data by sulcation index (SI). Both the mean GI and SI increased from GW 16, and were closely correlated with the cortical volume and the cortical surface area during fetal periods (their correlation coefficients marked more than 0.95). After birth, both the mean GI and SI decreased slightly by 2years of age, whereas the cortical volume and surface area continuously increased. Notably, histological analysis showed that the outer subventricular zone (oSVZ) in non-sulcal regions was thicker than that in the presumptive calcarine sulcal region on GW 13, preceding the infolding of the calcarine sulcus. The present results showed definite sulcal infolding on the cerebral cortical surface of the marmosets, with similar pattern and sequence of their emergences to other higher-order primates such as macaques and humans. Differential expansion of the oSVZ may be involved in gyral convolution and sulcal infolding in the developing cerebrum. © 2013 IBRO.

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Distinct roles of endogenous vascular endothelial factor receptor 1 and 2 in neural protection after spinal cord injury

Shinozaki Munehisa, Nakamura Masaya, Konomi Tsunehiko, Kobayashi Yoshiomi, Takano Morito, Saito Nobuhito, Toyama Yoshiaki, Okano Hideyuki

Neuroscience Research 78 （ 1 ） 55 - 64 2014年01月

ISSN 0168-0102

Secondary degeneration after spinal cord injury (SCI) is caused by increased vascular permeability, infiltration of inflammatory cells, and subsequent focal edema. Therapeutic interventions using neurotrophic factors have focused on the prevention of such reactions to reduce cell death and promote tissue regeneration. Vascular endothelial growth factor (VEGF) is a potent angiogenic and vascular permeability factor. However, the effect of VEGF on SCI remains controversial. VEGF signaling is primarily regulated through two primary receptors, VEGF receptor 1 (VEGF-R1) and VEGF receptor 2 (VEGF-R2). The purpose of this study was to examine the effects of intraperitoneal administration of VEGF-R1- and VEGF-R2-neutralizing antibodies on a mouse model of SCI. VEGF-R1 blockade, but not VEGF-R2 blockade, decreased the permeability and infiltration of inflammatory cells, and VEGF-R2 blockade caused a significant increase in neuronal apoptosis in the acute phase of SCI. VEGF-R2 blockade decreased the residual tissue area and the number of neural fibers in the chronic phase of SCI. VEGF-R2 blockade worsened the functional recovery and prolonged the latency of motor evoked potentials. These data suggest that endogenous VEGF-R2 plays a crucial role in neuronal protection after SCI. © 2013 Elsevier Ireland Ltd and the Japan Neuroscience Society.

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Generation of Germ Cells In Vitro in the Era of Induced Pluripotent Stem Cells

岡野栄之

MOLECULAR REPRODUCTION AND DEVELOPMENT 81 （ 1 ） 2 - 19 2014年01月

研究論文（学術雑誌）, ISSN 1040-452X

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Common marmoset as a new model animal for neuroscience research and genome editing technology

岡野栄之

DEVELOPMENT GROWTH & DIFFERENTIATION 56 （ 1 ） 53 - 62 2014年01月

研究論文（学術雑誌）, ISSN 0012-1592

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The miR-17/106-p38 axis is a key regulator of the neurogenic-to-gliogenic transition in developing neural stem/progenitor cells

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 111 （ 4 ） 1604 - 1609 2014年01月

研究論文（学術雑誌）, ISSN 0027-8424

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LIM Homeobox 8 (Lhx8) Is a Key Regulator of the Cholinergic Neuronal Function via a Tropomyosin Receptor Kinase A (TrkA)-mediated Positive Feedback Loop

岡野栄之

JOURNAL OF BIOLOGICAL CHEMISTRY 289 （ 2 ） 1000 - 1010 2014年01月

研究論文（学術雑誌）, ISSN 0021-9258

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Increased L1 Retrotransposition in the Neuronal Genome in Schizophrenia

岡野栄之

NEURON 81 （ 2 ） 306 - 313 2014年01月

研究論文（学術雑誌）, ISSN 0896-6273

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The RNA-binding Protein HuD Regulates Autophagosome Formation in Pancreatic beta Cells by Promoting Autophagy-related Gene 5 Expression

岡野栄之

JOURNAL OF BIOLOGICAL CHEMISTRY 289 （ 1 ） 112 - 121 2014年01月

研究論文（学術雑誌）, ISSN 0021-9258

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Role of opioid systems in the differentiation potential of pluripotent stem cells

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 124 52P - 52P 2014年

研究論文（学術雑誌）, ISSN 1347-8613

Transplantation of Neural Stem/Progenitor Cells at Different Locations in Mice With Spinal Cord Injury

岡野栄之

CELL TRANSPLANTATION 23 （ 11 ） 1451 - 1464 2014年

研究論文（学術雑誌）, ISSN 0963-6897

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Micro-RNA 200b/429 in the nucleus accumbens plays a key role in the neuropathic pain

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 124 101P - 101P 2014年

研究論文（学術雑誌）, ISSN 1347-8613

Possibility of a novel therapy for neuropathic pain by specific histone modification of astrocytic chemokines in the spinal cord

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 124 100P - 100P 2014年

研究論文（学術雑誌）, ISSN 1347-8613

Neural differentiation potentials in human embryonic stem cell lines

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 124 94P - 94P 2014年

研究論文（学術雑誌）, ISSN 1347-8613

Chronic itch requires STAT3-dependent reactive astrocytes in the spinal dorsal horn

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 124 81P - 81P 2014年

研究論文（学術雑誌）, ISSN 1347-8613

Analysis of glia-neuron interaction by using opto-fMRI

岡野栄之

JOURNAL OF PHARMACOLOGICAL SCIENCES 124 43P - 43P 2014年

研究論文（学術雑誌）, ISSN 1347-8613

I2020T mutant LRRK2 iPSC-derived neurons in the Sagamihara family exhibit increased Tau phosphorylation through the AKT/GSK-3 beta signaling pathway

岡野栄之

HUMAN MOLECULAR GENETICS 24 （ 17 ） 4879 - 4900 2015年09月

研究論文（学術雑誌）, ISSN 0964-6906

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Histological and electrophysiological analysis of the corticospinal pathway to forelimb motoneurons in common marmosets

岡野栄之

NEUROSCIENCE RESEARCH 98 35 - 44 2015年09月

研究論文（学術雑誌）, ISSN 0168-0102

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MicroRNA-153 Regulates the Acquisition of Gliogenic Competence by Neural Stem Cells

岡野栄之

STEM CELL REPORTS 5 （ 3 ） 365 - 377 2015年09月

研究論文（学術雑誌）, ISSN 2213-6711

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A new minimally-invasive method for microinjection into the mouse spinal dorsal horn

岡野栄之

SCIENTIFIC REPORTS 5 101P - 101P 2015年09月

研究論文（学術雑誌）, ISSN 2045-2322

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Acute reduction of neuronal RNA binding Elavl2 protein and Gap43 mRNA in mouse hippocampus after kainic acid treatment

Ohtsuka Takafumi, Yano Masato, Okano Hideyuki

Biochemical and Biophysical Research Communications 466 （ 1 ） 46 - 51 2015年08月

ISSN 0006-291X

Activity-dependent gene regulation in neurons has been hypothesized to be under transcriptional control and to include dramatic increases in immediate early genes (IEGs) after neuronal activity. In addition, several reports have focused on post-transcriptional regulation, which could be mediated by neuronal post-transcriptional regulators, including RNA binding proteins (RNABPs). One such protein family is the neuronal Elavls (nElavls; Elavl2, Elavl3, and Elavl4), whose members are widely expressed in peripheral and central nervous system. Previous reports showed that Elavl3 and 4 are up-regulated following repeated stimulation such as during cocaine administration, a seizure, or a spatial discrimination task. In this study, we focused on Elavl2, a candidate gene for schizophrenia and studied its role in neuronal activity. First we found that Elavl2 has a cell-type specific expression pattern that is highly expressed in hippocampal CA3 pyramidal neurons and hilar interneurons using Elavl2 specific antibody. Second, unexpectedly, we discovered that the Elavl2 protein level in the hippocampus was acutely down-regulated for 3 h after a kainic acid (KA)-induced seizure in the hippocampal CA3 region. In addition, level of Gap43 mRNA, a target mRNA of Elavl2 is decreased 12 h after KA treatment, thus suggesting the involvement of Elavl2 in activity-dependent RNA regulation.

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Voxel-based morphometry of the marmoset brain

Hikishima K., Ando K., Komaki Y., Kawai K., Yano R., Inoue T., Itoh T., Yamada M., Momoshima S., Okano H. J., Okano H.

Neuroscience 300 585 - 592 2015年08月

ISSN 0306-4522

Movement dysfunction in Parkinson's disease (PD) is caused by the degeneration of dopaminergic (DA) neurons in the substantia nigra (SN). Here, we established a method for voxel-based morphometry (VBM) and automatic tissue segmentation of the marmoset monkey brain using a 7-T animal scanner and applied the method to assess DA degeneration in a PD model, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated animals, with tyrosine-hydroxylase staining. The most significant decreases of local tissue volume were detected in the bilateral SN of MPTP-treated marmoset brains (-53.0% in right and -46.5% in left) and corresponded with the location of DA neurodegeneration found in histology (-65.4% in right). In addition to the SN, the decreases were also confirmed in the locus coeruleus, and lateral hypothalamus. VBM using 7-T MRI was effective in detecting volume loss in the SN of the PD-model marmoset. This study provides a potential basis for the application of VBM with ultra-high field MRI in the clinical diagnosis of PD. The developed method may also offer value in automatic whole-brain evaluation of structural changes for the marmoset monkey.

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q-space MR imaging of gastric carcinoma ex vivo

Yamada Ichiro, Hikishima Keigo, Miyasaka Naoyuki, Kato Keiji, Ito Eisaku, Kojima Kazuyuki, Kawano Tatsuyuki, Kobayashi Daisuke, Eishi Yoshinobu, Okano Hideyuki

Magnetic Resonance in Medicine 76 （ 2 ） 602 - 612 2015年08月

研究論文（学術雑誌）, 共著, ISSN 0740-3194

Purpose: The purpose of this study was to establish the feasibility of q-space imaging (QSI) as a method of assessing the depth of mural invasion, histologic grade, and the presence of lymph node metastasis in gastric carcinomas. Methods: A 7.0 Tesla MR imaging system was used to investigate 20 gastric specimens containing a carcinoma. QSI was performed by using the following parameters: 50-60 mm × 25-30 mm field of view, 2-mm section thickness, 256 × 128 matrix, 10 b values in the 0-7163 s/mm2 range, which corresponded to q values of 0-1026/cm, and motion-probing gradients perpendicular to the gastric wall. The MR images and the histopathologic findings were then compared. Results: The depth of tumor invasion of the gastric wall in all 20 carcinomas (100%) was established by using mean displacement, zero-displacement probability, and kurtosis maps. The QSI parameters were significantly correlated with the histologic grades of the gastric carcinomas (all P<0.001). The QSI parameters made it possible to differentiate between metastatic and nonmetastatic lymph nodes (all P=0.001). Conclusion: Ex vivo QSI facilitates excellent diagnostics for evaluating gastric carcinomas in terms of mural invasion, histologic grade, and the presence of lymph node metastasis.

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Migration and differentiation of transplanted enteric neural crest-derived cells in murine model of Hirschsprung's disease

岡野栄之

CYTOTECHNOLOGY 67 （ 4 ） 661 - 670 2015年08月

研究論文（学術雑誌）, ISSN 0920-9069

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CD34 and CD49f Double-Positive and Lineage Marker-Negative Cells Isolated from Human Myometrium Exhibit Stem Cell-Like Properties Involved in Pregnancy-Induced Uterine Remodeling

岡野栄之

BIOLOGY OF REPRODUCTION 93 （ 2 ） 17163 - 17168 2015年08月

研究論文（学術雑誌）, ISSN 0006-3363

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Engrafted Neural Stem/Progenitor Cells Promote Functional Recovery through Synapse Reorganization with Spared Host Neurons after Spinal Cord Injury

岡野栄之

STEM CELL REPORTS 5 （ 2 ） 264 - 277 2015年08月

研究論文（学術雑誌）, ISSN 2213-6711

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BDNF Induced by Treadmill Training Contributes to the Suppression of Spasticity and Allodynia After Spinal Cord Injury via Upregulation of KCC2

岡野栄之

NEUROREHABILITATION AND NEURAL REPAIR 29 （ 7 ） 677 - 689 2015年08月

研究論文（学術雑誌）, ISSN 1545-9683

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Neural stem/progenitor cell-laden microfibers promote transplant survival in a mouse transected spinal cord injury model

Sugai Keiko, Nishimura Soraya, Kato-Negishi Midori, Onoe Hiroaki, Iwanaga Shintaroh, Toyama Yoshiaki, Matsumoto Morio, Takeuchi Shoji, Okano Hideyuki, Nakamura Masaya

Journal of Neuroscience Research 93 （ 12 ） 1826 - 1838 2015年08月

共著, ISSN 0360-4012

Previous studies have demonstrated that transplantation of neural stem/progenitor cells (NS/PCs) into the lesioned spinal cord can promote functional recovery following incomplete spinal cord injury (SCI) in animal models. However, this strategy is insufficient following complete SCI because of the gap at the lesion epicenter. To obtain functional recovery in a mouse model of complete SCI, this study uses a novel collagen-based microfiber as a scaffold for engrafted NS/PCs. We hypothesized that the NS/PC-microfiber combination would facilitate lesion closure as well as transplant survival in the transected spinal cord. NS/PCs were seeded inside the novel microfibers, where they maintained their capacity to differentiate and proliferate. After transplantation, the stumps of the transected spinal cord were successfully bridged by the NS/PC-laden microfibers. Moreover, the transplanted cells migrated into the host spinal cord and differentiated into three neural lineages (astrocytes, neurons, and oligodendrocytes). However, the NS/PC-laden scaffold could not achieve a neural connection between the rostral end of the injury and the intact caudal area of the spinal cord, nor could it achieve recovery of motor function. To obtain optimal functional recovery, a microfiber design with a modified composition may be useful. Furthermore, combinatorial therapy with rehabilitation and/or medications should also be considered for practical success of biomaterial/cell transplantation-based approaches to regenerative medicine.

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Utility of Scalp Hair Follicles as a Novel Source of Biomarker Genes for Psychiatric Illnesses

岡野栄之

BIOLOGICAL PSYCHIATRY 78 （ 2 ） 116 - 125 2015年07月

研究論文（学術雑誌）, ISSN 0006-3223

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Allogeneic Neural Stem/Progenitor Cells Derived From Embryonic Stem Cells Promote Functional Recovery After Transplantation Into Injured Spinal Cord of Nonhuman Primates

岡野栄之

STEM CELLS TRANSLATIONAL MEDICINE 4 （ 7 ） 708 - 719 2015年07月

研究論文（学術雑誌）, ISSN 2157-6564

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Esophageal carcinoma: Evaluation with q-space diffusion-weighted MR imaging ex vivo

岡野栄之

MAGNETIC RESONANCE IN MEDICINE 73 （ 6 ） 2262 - 2273 2015年06月

研究論文（学術雑誌）, ISSN 0740-3194

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Gastric Carcinoma: Ex Vivo MR Imaging at 7.0 T-Correlation with Histopathologic Findings

岡野栄之

RADIOLOGY 275 （ 3 ） 841 - 848 2015年06月

研究論文（学術雑誌）, ISSN 0033-8419

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Brains, Genes, and Primates

岡野栄之

NEURON 86 （ 3 ） 617 - 631 2015年05月

研究論文（学術雑誌）, ISSN 0896-6273

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Differentiation of multipotent neural stem cells derived from Rett syndrome patients is biased toward the astrocytic lineage

岡野栄之

MOLECULAR BRAIN 27 （ 8(1) ） 31 2015年05月

研究論文（学術雑誌）, ISSN 1756-6606

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Dynamic regulation of innate immune responses in Drosophila by Senju-mediated glycosylation

岡野栄之

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 112 （ 18 ） 5809 - 5814 2015年05月

研究論文（学術雑誌）, ISSN 0027-8424

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Parkinson Disease: Diffusion MR Imaging to Detect Nigrostriatal Pathway Loss in a Marmoset Model Treated with 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine

岡野栄之

RADIOLOGY 275 （ 2 ） 430 - 437 2015年05月

研究論文（学術雑誌）, ISSN 0033-8419

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Disrupted-in-schizophrenia 1 regulates transport of ITPR1 mRNA for synaptic plasticity

岡野栄之

NATURE NEUROSCIENCE 18 （ 5 ） 698 - + 2015年05月

研究論文（学術雑誌）, ISSN 1097-6256

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Brain/MINDS: brain-mapping project in Japan

岡野栄之

PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES 370 （ 1668 ） 33 - 41 2015年05月

研究論文（学術雑誌）, ISSN 0962-8436

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Ultra-high-resolution MR imaging of esophageal carcinoma at ultra-high field strength (7.0 T) ex vivo: correlation with histopathologic findings

岡野栄之

MAGNETIC RESONANCE IMAGING 33 （ 4 ） 413 - 419 2015年05月

研究論文（学術雑誌）, ISSN 0730-725X

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Marmoset neuroscience

岡野栄之

NEUROSCIENCE RESEARCH 93 1 - 2 2015年04月

研究論文（学術雑誌）, ISSN 0168-0102

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Brain-mapping projects using the common marmoset

岡野栄之

NEUROSCIENCE RESEARCH 93 3 - 7 2015年04月

研究論文（学術雑誌）, ISSN 0168-0102

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Notch Inhibition Induces Cochlear Hair Cell Regeneration and Recovery of Hearing after Acoustic Trauma (vol 77, pg 58, 2013)

岡野栄之

NEURON 86 （ 1 ） S347 - S348 2015年04月

研究論文（学術雑誌）, ISSN 0896-6273

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Gene expression ontogeny of spermatogenesis in the marmoset uncovers primate characteristics during testicular development

岡野栄之

DEVELOPMENTAL BIOLOGY 400 （ 1 ） 43 - 58 2015年04月

研究論文（学術雑誌）, ISSN 0012-1606

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Alternative Role of HuD Splicing Variants in Neuronal Differentiation

岡野栄之

JOURNAL OF NEUROSCIENCE RESEARCH 93 （ 3 ） 399 - 409 2015年03月

研究論文（学術雑誌）, ISSN 0360-4012

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Manipulating cell fate in the cochlea: a feasible therapy for hearing loss

岡野栄之

TRENDS IN NEUROSCIENCES 38 （ 3 ） 139 - 144 2015年03月

研究論文（学術雑誌）, ISSN 0166-2236

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Long-Term Safety Issues of iPSC-Based Cell Therapy in a Spinal Cord Injury Model: Oncogenic Transformation with Epithelial-Mesenchymal Transition

岡野栄之

STEM CELL REPORTS 4 （ 3 ） 360 - 373 2015年03月

研究論文（学術雑誌）, ISSN 2213-6711

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Optogenetic Activation of CA1 Pyramidal Neurons at the Dorsal and Ventral Hippocampus Evokes Distinct Brain-Wide Responses Revealed by Mouse fMRI

岡野栄之

PLOS ONE 10 （ 3 ） 12829 - 12838 2015年03月

研究論文（学術雑誌）, ISSN 1932-6203

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Novel Overgrowth Syndrome Phenotype Due to Recurrent De Novo PDGFRB Mutation

岡野栄之

JOURNAL OF PEDIATRICS 166 （ 2 ） 483 - 486 2015年02月

研究論文（学術雑誌）, ISSN 0022-3476

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Connectomics: comprehensive approaches for whole-brain mapping

岡野栄之

MICROSCOPY 64 （ 1 ） 57 - 67 2015年02月

研究論文（学術雑誌）, ISSN 0022-0744

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Controlling Immune Rejection Is a Fail-Safe System against Potential Tumorigenicity after Human iPSC-Derived Neural Stem Cell Transplantation

岡野栄之

PLOS ONE 10 （ 2 ） 286 - 286 2015年02月

研究論文（学術雑誌）, ISSN 1932-6203

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RNA-binding protein research with transcriptome-wide technologies in neural development

岡野栄之

CELL AND TISSUE RESEARCH 359 （ 1 ） 135 - 144 2015年01月

研究論文（学術雑誌）, ISSN 0302-766X

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Involvement of ER Stress in Dysmyelination of Pelizaeus-Merzbacher Disease with PLP1 Missense Mutations Shown by iPSC-Derived Oligodendrocytes (vol 2, pg 648, 2014)

岡野栄之

STEM CELL REPORTS 4 （ 1 ） 170 - 170 2015年01月

研究論文（学術雑誌）, ISSN 2213-6711

Clinical utility of neuronal cells directly converted from fibroblasts of patients for neuropsychiatric disorders

Kano S., Yuan M., Cardarelli R. A., Maegawa G., Higurashi N., Gaval-Cruz M., Wilson A. M., Tristan C., Kondo M. A., Chen Y., Koga M., Obie C., Ishizuka K., Seshadri S., Srivastava R., Kato T. A., Horiuchi Y., Sedlak T. W., Lee Y., Rapoport J. L., Hirose S., Okano H., Valle D., O’Donnell P., Sawa A., Kai M.

Current Molecular Medicine 15 （ 2 ） 138 - 145 2015年

ISSN 1566-5240

Methodologies for generating functional neuronal cells directly from human fibroblasts [induced neuronal (iN) cells] have been recently developed, but the research so far has only focused on technical refinements or recapitulation of known pathological phenotypes. A critical question is whether this novel technology will contribute to elucidation of novel disease mechanisms or evaluation of therapeutic strategies. Here we have addressed this question by studying Tay-Sachs disease, a representative lysosomal storage disease, and Dravet syndrome, a form of severe myoclonic epilepsy in infancy, using human iN cells with feature of immature postmitotic glutamatergic neuronal cells. In Tay-Sachs disease, we have successfully characterized canonical neuronal pathology, massive accumulation of GM2 ganglioside, and demonstrated the suitability of this novel cell culture for future drug screening. In Dravet syndrome, we have identified a novel functional phenotype that was not suggested by studies of classical mouse models and human autopsied brains. Taken together, the present study demonstrates that human iN cells are useful for translational neuroscience research to explore novel disease mechanisms and evaluate therapeutic compounds. In the future, research using human iN cells with well-characterized genomic landscape can be integrated into multidisciplinary patient-oriented research on neuropsychiatric disorders to address novel disease mechanisms and evaluate therapeutic strategies.

Control of the Survival and Growth of Human Glioblastoma Grafted Into the Spinal Cord of Mice by Taking Advantage of Immunorejection

岡野栄之

CELL TRANSPLANTATION 24 （ 7 ） 1299 - 1311 2015年

研究論文（学術雑誌）, ISSN 0963-6897

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atlasBREX: Automated template-derived brain extraction in animal MRI

Lohmeier J., Kaneko T., Hamm B., Makowski M., Okano H.

Scientific Reports （Scientific Reports） 9 （ 1 ） 2019年12月

© 2019, The Author(s). We proposed a generic template-derived approach for (semi-) automated brain extraction in animal MRI studies and evaluated our implementation with different animal models (macaque, marmoset, rodent) and MRI protocols (T1, T2). While conventional MR-neuroimaging studies perform brain extraction as an initial step priming subsequent image-registration from subject to template, our proposed approach propagates an anatomical template to (whole-head) individual subjects in reverse order, which is challenging due to the surrounding extracranial tissue, greater differences in contrast pattern and larger areas with field inhomogeneity. As a novel approach, the herein introduced brain extraction algorithm derives whole-brain segmentation using rigid and non-rigid deformation based on unbiased anatomical atlas building with a priori estimates from study-cohort and an initial approximate brain extraction. We evaluated our proposed method in comparison to several other technical approaches including “Marker based watershed scalper”, “Brain-Extraction-Tool”, “3dSkullStrip”, “Primatologist-Toolbox”, “Rapid Automatic Tissue Segmentation” and “Robust automatic rodent brain extraction using 3D pulse-coupled neural networks” with manual skull-stripping as reference standard. ABX demonstrated best performance with accurate (≥92%) and consistent results throughout datasets and across species, age and MRI protocols. ABX was made available to the public with documentation, templates and sample material (https://www.github.com/jlohmeier/atlasBREX).

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The adeno-associated virus rh10 vector is an effective gene transfer system for chronic spinal cord injury

Hoshino Y., Nishide K., Nagoshi N., Shibata S., Moritoki N., Kojima K., Tsuji O., Matsumoto M., Kohyama J., Nakamura M., Okano H.

Scientific Reports （Scientific Reports） 9 （ 1 ） 2019年12月

© 2019, The Author(s). Treatment options for chronic spinal cord injury (SCI) remain limited due to unfavourable changes in the microenvironment. Gene therapy can overcome these barriers through continuous delivery of therapeutic gene products to the target tissue. In particular, adeno-associated virus (AAV) vectors are potential candidates for use in chronic SCI, considering their safety and stable gene expression in vivo. Given that different AAV serotypes display different cellular tropisms, it is extremely important to select an optimal serotype for establishing a gene transfer system during the chronic phase of SCI. Therefore, we generated multiple AAV serotypes expressing ffLuc-cp156, a fusion protein of firefly luciferase and Venus, a variant of yellow fluorescent protein with fast and efficient maturation, as a reporter, and we performed intraparenchymal injection in a chronic SCI mouse model. Among the various serotypes tested, AAVrh10 displayed the highest photon count on bioluminescence imaging. Immunohistological analysis revealed that AAVrh10 showed favourable tropism for neurons, astrocytes, and oligodendrocytes. Additionally, with AAVrh10, the area expressing Venus was larger in the injury epicentre and extended to the surrounding tissue. Furthermore, the fluorescence intensity was significantly higher with AAVrh10 than with the other vectors. These results indicate that AAVrh10 may be an appropriate serotype for gene delivery to the chronically injured spinal cord. This promising tool may be applied for research and development related to the treatment of chronic SCI.

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Role of cyclooxygenase-2-mediated prostaglandin E2-prostaglandin E receptor 4 signaling in cardiac reprogramming

Muraoka N., Nara K., Tamura F., Kojima H., Yamakawa H., Sadahiro T., Miyamoto K., Isomi M., Haginiwa S., Tani H., Kurotsu S., Osakabe R., Torii S., Shimizu S., Okano H., Sugimoto Y., Fukuda K., Ieda M.

Nature Communications （Nature Communications） 10 （ 1 ） 2019年12月

© 2019, The Author(s). Direct cardiac reprogramming from fibroblasts can be a promising approach for disease modeling, drug screening, and cardiac regeneration in pediatric and adult patients. However, postnatal and adult fibroblasts are less efficient for reprogramming compared with embryonic fibroblasts, and barriers to cardiac reprogramming associated with aging remain undetermined. In this study, we screened 8400 chemical compounds and found that diclofenac sodium (diclofenac), a non-steroidal anti-inflammatory drug, greatly enhanced cardiac reprogramming in combination with Gata4, Mef2c, and Tbx5 (GMT) or GMT plus Hand2. Intriguingly, diclofenac promoted cardiac reprogramming in mouse postnatal and adult tail-tip fibroblasts (TTFs), but not in mouse embryonic fibroblasts (MEFs). Mechanistically, diclofenac enhanced cardiac reprogramming by inhibiting cyclooxygenase-2, prostaglandin E2/prostaglandin E receptor 4, cyclic AMP/protein kinase A, and interleukin 1β signaling and by silencing inflammatory and fibroblast programs, which were activated in postnatal and adult TTFs. Thus, anti-inflammation represents a new target for cardiac reprogramming associated with aging.

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Robust and efficient knock-in in embryonic stem cells and early-stage embryos of the common marmoset using the CRISPR-Cas9 system

Yoshimatsu S., Okahara J., Sone T., Takeda Y., Nakamura M., Sasaki E., Kishi N., Shiozawa S., Okano H.

Scientific Reports （Scientific Reports） 9 （ 1 ） 2019年12月

© 2019, The Author(s). Genome editing technology greatly facilitates the genetic modification of various cells and animals. The common marmoset (Callithrix jacchus), a small non-human primate which exhibits high reproductive efficiency, is a widely used animal model in biomedical research. Developing genome editing techniques in the common marmoset will further enhance its utility. Here, we report the successful establishment of a knock-in (KI) method for marmoset embryonic stem cells (ESCs), which is based on the CRISPR-Cas9 system. The use of CRISPR-Cas9, mediated by homologous recombination (HR), enhanced the KI efficiency in marmoset ESCs. Furthermore, we succeeded in performing KI in early-stage marmoset embryos. In the course of the experiments, we found that HR in the marmoset ESCs is innately highly efficient. This suggested that the marmoset possesses a repair mechanism for DNA double-strand breaks. The current study will facilitate the generation of genetically modified marmosets and gene function analysis in the marmoset.

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Dual usage of a stage-specific fluorescent reporter system based on a helper-dependent adenoviral vector to visualize osteogenic differentiation

Sone T., Shin M., Ouchi T., Sasanuma H., Miyamoto A., Ohte S., Tsukamoto S., Nakanishi M., Okano H., Katagiri T., Mitani K.

Scientific Reports （Scientific Reports） 9 （ 1 ） 2019年12月

© 2019, The Author(s). We developed a reporter system that can be used in a dual manner in visualizing mature osteoblast formation. The system is based on a helper-dependent adenoviral vector (HDAdV), in which a fluorescent protein, Venus, is expressed under the control of the 19-kb human osteocalcin (OC) genomic locus. By infecting human and murine primary osteoblast (POB) cultures with this reporter vector, the cells forming bone-like nodules were specifically visualized by the reporter. In addition, the same vector was utilized to efficiently knock-in the reporter into the endogenous OC gene of human induced pluripotent stem cells (iPSCs), by homologous recombination. Neural crest-like cells (NCLCs) derived from the knock-in reporter iPSCs were differentiated into osteoblasts forming bone-like nodules and could be visualized by the expression of the fluorescent reporter. Living mature osteoblasts were then isolated from the murine mixed POB culture by fluorescence-activated cell sorting (FACS), and their mRNA expression profile was analyzed. Our study presents unique utility of reporter HDAdVs in stem cell biology and related applications.

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Cell therapy for spinal cord injury using induced pluripotent stem cells

Nagoshi N., Tsuji O., Nakamura M., Okano H.

Regenerative Therapy （Regenerative Therapy） 11 75 - 80 2019年12月

© 2019 The Japanese Society for Regenerative Medicine For the past few decades, spinal cord injury (SCI) has been believed to be an incurable traumatic condition, but with recent developments in stem cell biology, the field of regenerative medicine has gained hopeful momentum in the development of a treatment for this challenging pathology. Among the treatment candidates, transplantation of neural precursor cells has gained remarkable attention as a reasonable therapeutic intervention to replace the damaged central nervous system cells and promote functional recovery. Here, we highlight transplantation therapy techniques using induced pluripotent stem cells to treat SCI and review the recent research giving consideration to future clinical applications.

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Ropinirole hydrochloride remedy for amyotrophic lateral sclerosis – Protocol for a randomized, double-blind, placebo-controlled, single-center, and open-label continuation phase I/IIa clinical trial (ROPALS trial)

Morimoto S., Takahashi S., Fukushima K., Saya H., Suzuki N., Aoki M., Okano H., Nakahara J.

Regenerative Therapy （Regenerative Therapy） 11 143 - 166 2019年12月

© 2019 The Japanese Society for Regenerative Medicine Introduction: Amyotrophic lateral sclerosis (ALS) is an intractable and incurable neurological disease. It is a progressive disease characterized by muscle atrophy and weakness caused by selective vulnerability of upper and lower motor neurons. In disease research, it has been common to use mouse models carrying mutations in responsible genes for familial ALS as pathological models of ALS. However, there is no model that has reproduced the actual conditions of human spinal cord pathology. Thus, we developed a method of producing human spinal motor neurons using human induced pluripotent stem cells (iPSCs) and an innovative experimental technique for drug screening. As a result, ropinirole hydrochloride was eventually discovered after considering such results as its preferable transitivity in the brain and tolerability, including possible adverse reactions. Therefore, we explore the safety, tolerability and efficacy of ropinirole hydrochloride as an ALS treatment in this clinical trial. Methods: The ROPALS trial is a single-center double-blind randomized parallel group-controlled trial of the safety, tolerability, and efficacy of the ropinirole hydrochloride extended-release tablet (Requip CR) at 2- to 16-mg doses in patients with ALS. Twenty patients will be recruited for the active drug group (fifteen patients) and placebo group (five patients). All patients will be able to receive the standard ALS treatment of riluzole if not changed the dosage during this trial. The primary outcome will be safety and tolerability at 24 weeks, defined from the date of randomization. Secondary outcome will be the efficacy, including any change in the ALS Functional Rating Scale-Revised (ALSFRS-R), change in the Combined Assessment of Function and Survival (CAFS), and the composite endpoint as a sum of Z-transformed scores on various clinical items. Notably, we will perform an explorative search for a drug effect evaluation using the patient-derived iPSCs to prove this trial concept. Eligible patients will have El Escorial Possible, clinically possible and laboratory-supported, clinically probable, or clinically definite amyotrophic lateral sclerosis with disease duration less than 60 months (inclusive), an ALSFRS-R score ≥2 points on all items and age from 20 to 80 years. Conclusion: Patient recruitment began in December 2018 and the last patient is expected to complete the trial protocol in November 2020. Trial registration: Current controlled trials UMIN000034954 and JMA-IIA00397 Protocol version: version 1.6 (Date; 5/Apr/2019).

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Generation of a human iPS cell line (CGMH.SLC26A4919-2) from a Pendred syndrome patient carrying SLC26A4 c.919-2A>G splice-site mutation

Cheng Y., Chan Y., Hu C., Lu Y., Saeki T., Hosoya M., Saegusa C., Fujioka M., Okano H., Weng S., Hsu C., Chang K., Wu C.

Stem Cell Research （Stem Cell Research） 40 2019年10月

ISSN 18735061

© 2019 SLC26A4 is the second most frequent gene implicated in congenital hearing loss after GJB2 mutations. Here, we report the generation of induced pluripotent stem cells (iPSCs), from a patient who was carrying a homozygous c.919-2A>G variant in the SLC26A4 gene. This is the most common variant of SLC26A4 gene in the Chinese population and the second most prevalent one in other Asian countries. The established patient-derived iPSC displayed all the features of pluripotent stem cell markers and had the ability to differentiate into all of the three germ layers and possessed a normal karyotype.

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Analysis of skeletal-muscle condition after excessive loading of the lower legs by sequential magnetic resonance imaging

Hata J., Endo K., Tsuji O., Arakawa S., Sato M., Yagi K., Fujiyoshi K., Okano H., Nakamura M.

Journal of Orthopaedic Science （Journal of Orthopaedic Science） 24 （ 5 ） 873 - 880 2019年09月

ISSN 09492658

© 2019 Objectives: To determine the changes of muscle conditions of lower leg after stretch shortening cycle exercises by MRI. Subjects and methods: This study included 20 healthy male adult volunteers. After conducting control MRI, each subject performed 3 sets of exercise loading, each set consisting of 100 repetitions of rebound jumping on one foot. MRIs were performed immediately after exercise loading (0 h), 6, 30, and 168 h later. After constructing T1/T2/Fractional Anisotropy (FA)/Apparent Diffusion Coefficient (ADC) maps, the changes of three skeletal muscles of the leg (the tibialis anterior [TA], soleus [SOL], and gastrocnemius [GA]) were quantitatively evaluated in each map at each time point. Results: The T1 and T2 values were prolonged after exercise loading, and there was a delay in the recovery of T1 at 6 and 30 h after exercise loading, as compared to those of T2 values over time. The ADC values were elevated in all three muscles immediately after exercise loading, then recovered more slowly than T1 and T2, and still had not returned to baseline 168 h after exercise loading. The FA value decreased in all three muscles after exercise loading, with the greatest decrease occurring immediately after exercise loading. As with the ADC values, the FA values were slow to recover from the decrease, and had not returned to baseline levels 168 h post-loading. Conclusion: The delay of T1 value recovery suggested that the T1 value may reflect the muscle condition like fatigue and damage. Changes in the ADC and FA values over time suggested that structural changes such as minute muscular injuries can be detected by diffusion-weighted MRI. Meanwhile, the changes observed in the T1 and T2 values suggested that the measured relaxation time data reflected not only the water volume in the muscle, but also the muscle condition after exercise loading.

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Establishment of induced pluripotent stem cells from common marmoset fibroblasts by RNA-based reprogramming

Nakajima M., Yoshimatsu S., Sato T., Nakamura M., Okahara J., Sasaki E., Shiozawa S., Okano H.

Biochemical and Biophysical Research Communications （Biochemical and Biophysical Research Communications） 515 （ 4 ） 593 - 599 2019年08月

ISSN 0006291X

© 2019 Elsevier Inc. The common marmoset (marmoset; Callithrix jacchus) shows anatomical and physiological features that are in common with humans. Establishing induced pluripotent stem cells (iPSCs) from marmosets holds promise for enhancing the utility of the animal model for biomedical and preclinical studies. However, in spite of the presence of some previous reports on marmoset iPSCs, the reprogramming technology in marmosets is still under development. In particular, the efficacy of RNA-based reprogramming has not been thoroughly investigated. In this study, we attempted RNA-based reprogramming for deriving iPSCs from marmoset fibroblasts. Although we failed to derive iPSC colonies from marmoset fibroblasts by using a conventional RNA-based reprogramming method previously validated in human fibroblasts, we succeeded in deriving colony-forming cells with a modified induction medium supplemented with a novel set of small molecules. Importantly, following one-week culture of the colony-forming cells in conventional embryonic stem cell (ESC) medium, we obtained iPSCs which express endogenous pluripotent markers and show a differentiation potential into all three germ layers. Taken together, our results indicate that RNA-based reprogramming, which is valuable for deriving transgene-free iPSCs, is applicable to marmosets.

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Semi-supervised deep learning of brain tissue segmentation

Ito R., Nakae K., Hata J., Okano H., Ishii S.

Neural Networks （Neural Networks） 116 25 - 34 2019年08月

ISSN 08936080

© 2019 Elsevier Ltd Brain image segmentation is of great importance not only for clinical use but also for neuroscience research. Recent developments in deep neural networks (DNNs) have led to the application of DNNs to brain image segmentation, which required extensive human annotations of whole brain images. Annotating three-dimensional brain images requires laborious efforts by expert anatomists because of the differences among images in terms of their dimensionality, noise, contrast, or ambiguous boundaries that even prevent these experts from necessarily attaining consistency. This paper proposes a semi-supervised learning framework to train a DNN based on a relatively small number of annotated (labeled) images, named atlases, but also a relatively large number of unlabeled images by leveraging image registration to attach pseudo-labels to images that were originally unlabeled. We applied our proposed method to two different datasets: open human brain images and our original marmoset brain images. When provided with the same number of atlases for training, we found our method achieved superior and more stable segmentation results than those by existing registration-based and DNN-based methods.

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Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons

Akiyama T., Suzuki N., Ishikawa M., Fujimori K., Sone T., Kawada J., Funayama R., Fujishima F., Mitsuzawa S., Ikeda K., Ono H., Shijo T., Osana S., Shirota M., Nakagawa T., Kitajima Y., Nishiyama A., Izumi R., Morimoto S., Okada Y., Kamei T., Nishida M., Nogami M., Kaneda S., Ikeuchi Y., Mitsuhashi H., Nakayama K., Fujii T., Warita H., Okano H., Aoki M.

EBioMedicine （EBioMedicine） 45 362 - 378 2019年07月

© 2019 The Authors Background: The characteristic structure of motor neurons (MNs), particularly of the long axons, becomes damaged in the early stages of amyotrophic lateral sclerosis (ALS). However, the molecular pathophysiology of axonal degeneration remains to be fully elucidated. Method: Two sets of isogenic human-induced pluripotent stem cell (hiPSCs)-derived MNs possessing the single amino acid difference (p.H517D) in the fused in sarcoma (FUS) were constructed. By combining MN reporter lentivirus, MN specific phenotype was analyzed. Moreover, RNA profiling of isolated axons were conducted by applying the microfluidic devices that enable axon bundles to be produced for omics analysis. The relationship between the target gene, which was identified as a pathological candidate in ALS with RNA-sequencing, and the MN phenotype was confirmed by intervention with si-RNA or overexpression to hiPSCs-derived MNs and even in vivo. The commonality was further confirmed with other ALS-causative mutant hiPSCs-derived MNs and human pathology. Findings: We identified aberrant increasing of axon branchings in FUS-mutant hiPSCs-derived MN axons compared with isogenic controls as a novel phenotype. We identified increased level of Fos-B mRNA, the binding target of FUS, in FUS-mutant MNs. While Fos-B reduction using si-RNA or an inhibitor ameliorated the observed aberrant axon branching, Fos-B overexpression resulted in aberrant axon branching even in vivo. The commonality of those phenotypes was further confirmed with other ALS causative mutation than FUS. Interpretation: Analyzing the axonal fraction of hiPSC-derived MNs using microfluidic devices revealed that Fos-B is a key regulator of FUS-mutant axon branching. Fund: Japan Agency for Medical Research and development; Japanese Ministry of Education, Culture, Sports, Science and Technology Clinical Research, Innovation and Education Center, Tohoku University Hospital; Japan Intractable Diseases (Nanbyo) Research Foundation; the Kanae Foundation for the Promotion of Medical Science; and “Inochi-no-Iro” ALS research grant.

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Degradation of Extracellular Matrix by Matrix Metalloproteinase 2 Is Essential for the Establishment of the Blood-Brain Barrier in Drosophila

Kanda H., Shimamura R., Koizumi-Kitajima M., Okano H.

iScience （iScience） 16 218 - 229 2019年06月

© 2019 The Author(s) The blood-brain barrier (BBB) is an essential system that isolates the central nervous system from the internal environment. Increasing evidence has begun to reveal the molecules that are required for BBB integrity. However, how these components are regulated remains unclear. Here we report that a matrix metalloproteinase, Mmp2, is essential for the establishment of the BBB in Drosophila. In the absence of mmp2, the BBB becomes leaky, which allows the tracer to penetrate the brain. Moreover, the expression pattern of a junctional component, Neuroglian, is altered. We also find that the regulation of the amounts of particular extracellular matrix components is critical for BBB establishment. Furthermore, the process of mesenchymal-epithelial transition of BBB-forming cells is perturbed in the absence of Mmp2. These data indicate that the presence of Mmp(s), which is typically considered to be a risk factor for BBB degradation, is essential for BBB integrity in Drosophila. Biological Sciences; Molecular Biology; Cell Biology

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Involvement of p38 in Age-Related Decline in Adult Neurogenesis via Modulation of Wnt Signaling

Kase Y., Otsu K., Shimazaki T., Okano H.

Stem Cell Reports （Stem Cell Reports） 12 （ 6 ） 1313 - 1328 2019年06月

ISSN 22136711

© 2019 The Author(s) Neurogenesis in specific brain regions in adult mammals decreases with age. Progressive reduction in the proliferation of neural stem and progenitor cells (NS/PCs) is a primary cause of this age-associated decline. However, the mechanism responsible for this reduction is poorly understood. We identify p38 MAPK as a key factor in the proliferation of neural progenitor cells (NPCs) in adult neurogenic niches. p38 expression in adult NS/PCs is downregulated during aging. Deletion of p38α in NS/PCs specifically reduces the proliferation of NPCs but not stem cells. Conversely, forced expression of p38α in NS/PCs in the aged mouse subventricular zone (SVZ) restores NPC proliferation and neurogenesis, and prevents age-dependent SVZ atrophy. We also found that p38 is necessary for suppressing the expression of Wnt antagonists DKK1 and SFRP3, which inhibit the proliferation of NPCs. Age-related reduction in p38 thus leads to decreased adult neurogenesis via downregulation of Wnt signaling. Kase et al. show that p38 expression in neural stem and progenitor cells (NS/PCs) in the adult brain decreases with aging. This reduction specifically causes proliferation defect in neural progenitor cells (NPCs), leading to the age-dependent decline of adult neurogenesis. Conversely, overexpression of p38α in NS/PCs in the aged brain restores NPC proliferation without exhaustion of neural stem cells.

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Cell-specific overexpression of COMT in dopaminergic neurons of Parkinson's disease

Kuzumaki N., Suda Y., Iwasawa C., Narita M., Sone T., Watanabe M., Maekawa A., Matsumoto T., Akamatsu W., Igarashi K., Tamura H., Takeshima H., Tawfik V., Ushijima T., Hattori N., Okano H., Narita M.

Brain : a journal of neurology （Brain : a journal of neurology） 142 （ 6 ） 1675 - 1689 2019年06月

© The Author(s) (2019). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For Permissions, please email: journals.permissions@oup.com. The mechanism by which dopaminergic neurons are selectively affected in Parkinson's disease is not fully understood. In this study, we found a dramatic increase in the expression of catechol-O-methyltransferase (COMT), along with a lower level of DNA methylation, in induced pluripotent stem cell-derived dopaminergic neurons from patients with parkin (PARK2) gene mutations compared to those from healthy controls. In addition, a significant increase in the expression of COMT was found in dopaminergic neurons of isogenic PARK2 induced pluripotent stem cell lines that mimicked loss of function of PARK2 by CRISPR Cas9 technology. In dopamine transporter (DAT)-Cre mice, overexpression of COMT, specifically in dopaminergic neurons of the substantia nigra, produced cataleptic behaviours associated with impaired motor coordination. These findings suggest that upregulation of COMT, likely resulting from DNA hypomethylation, in dopaminergic neurons may contribute to the initial stage of neuronal dysfunction in Parkinson's disease.

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Estimating the concentration of therapeutic range using disease-specific iPS cells: Low-dose rapamycin therapy for Pendred syndrome

Hosoya M., Saeki T., Saegusa C., Matsunaga T., Okano H., Fujioka M., Ogawa K.

Regenerative Therapy （Regenerative Therapy） 10 54 - 63 2019年06月

© 2018 The Japanese Society for Regenerative Medicine Introduction: Pendred syndrome is an autosomal-recessive disease characterized by congenital hearing loss and thyroid goiter. Previously, cell stress susceptibilities were shown to increase in patient-derived cells with intracellular aggregation using an in vitro acute cochlear cell model derived from patient-specific pluripotent stem (iPS) cells. Moreover, we showed that rapamycin can relieve cell death. However, studies regarding long-term cell survival without cell stressors that mimic the natural course of disease or the rational minimum concentration of rapamycin that prevents cell death are missing. Methods: In this report, we first investigated the rational minimum concentration of rapamycin using patient-specific iPS cells derived-cochlear cells with three different conditions of acute stress. We next confirmed the effects of rapamycin in long-term cell survival and phenotypes by using cochlear cells derived from three different patient-derived iPS cells. Results: We found that inner ear cells derived from Pendred syndrome patients are more vulnerable than those from healthy individuals during long-term culturing; however, this susceptibility was relieved via treatment with low-dose rapamycin. The slow progression of hearing loss in patients may be explained, in part, by the vulnerability observed in patient cells during long-term culturing. We successfully evaluated the rational minimum concentration of rapamycin for treatment of Pendred syndrome. Conclusion: Our results suggest that low-dose rapamycin not only decreases acute symptoms but may prevent progression of hearing loss in Pendred syndrome patients.

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Visualization of spatiotemporal dynamics of human glioma stem cell invasion

Tamura R., Miyoshi H., Sampetrean O., Shinozaki M., Morimoto Y., Iwasawa C., Fukaya R., Mine Y., Masuda H., Maruyama T., Narita M., Saya H., Yoshida K., Okano H., Toda M.

Molecular Brain （Molecular Brain） 12 （ 1 ） 2019年05月

© 2019 The Author(s). Glioblastoma exhibits phenotypic and genetic heterogeneity, aggressive invasiveness, therapeutic resistance, and tumor recurrence, which can be explained by the existence of glioma stem cells (GSCs). In this study, we visualized the spatiotemporal dynamics of invasion of human GSCs in an orthotopic xenograft mouse model using time-lapse imaging of organotypic brain slice cultures and three-dimensional imaging of optically cleared whole brains. GSCs implanted in the striatum exhibited directional migration toward axon bundles, perivascular area, and the subventricular zone around the inferior horn of the lateral ventricle. GSCs migrated in a helical pattern around axon bundles in the striatum and invaded broadly in both the rostral and caudal directions. GSCs in the corpus callosum migrated more rapidly and unidirectionally toward the contralateral side with pseudopod extension. These characteristics of GSC invasion shared histological features observed in glioblastoma patients. Spatiotemporal visualization techniques can contribute to the elucidation of the mechanisms underlying GSC invasion that may lead to the development of effective therapy for glioblastoma.

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Correlative study using structural MRI and super-resolution microscopy to detect structural alterations induced by long-term optogenetic stimulation of striatal medium spiny neurons

Abe Y., Komaki Y., Seki F., Shibata S., Okano H., Tanaka K.

Neurochemistry International （Neurochemistry International） 125 163 - 174 2019年05月

ISSN 01970186

© 2019 Elsevier Ltd Striatal medium spiny neurons (MSNs) control motor function. Hyper- or hypo-activity of MSNs coincides with basal ganglia-related movement disorders. Based on the assumption that lasting alterations in neuronal activity lead to structural changes in the brain, understanding these structural alterations may be used to infer MSN functional abnormalities. To infer MSN function from structural data, understanding how long-lasting alterations in MSN activity affect brain morphology is essential. To address this, we utilized a simplified model of functional induction by stimulating MSNs expressing channelrhodopsin 2 (ChR2). Subsequent structural alterations which induced long-term activity changes in these MSNs were investigated in the striatal pathway and its associated regions by diffusion tensor imaging (DTI) and histological assessment with super-resolution microscopy. DTI detected changes in the striatum, substantia nigra, and motor cortex. Histological assessment found a reduction in the diameter of myelinated cortical axons as well as MSN dendrites and axons. The structural changes showed a high correlation between DTI parameters and histological data. These results demonstrated that long-term neural activation in the MSNs alters the diameter of MSN and cortical neurons fibers. This study provides a tool for understanding the causal relationship between functional and structural alterations.

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Large-area fluorescence and electron microscopic correlative imaging with multibeam scanning electron microscopy

Shibata S., Iseda T., Mitsuhashi T., Oka A., Shindo T., Moritoki N., Nagai T., Otsubo S., Inoue T., Sasaki E., Akazawa C., Takahashi T., Schalek R., Lichtman J., Okano H.

Frontiers in Neural Circuits （Frontiers in Neural Circuits） 13 2019年04月

ISSN 16625110

© 2019 Shibata, Iseda, Mitsuhashi, Oka, Shindo, Moritoki, Nagai, Otsubo, Inoue, Sasaki, Akazawa, Takahashi, Schalek, Lichtman and Okano. Recent improvements in correlative light and electron microscopy (CLEM) technology have led to dramatic improvements in the ability to observe tissues and cells. Fluorescence labeling has been used to visualize the localization of molecules of interest through immunostaining or genetic modification strategies for the identification of the molecular signatures of biological specimens. Newer technologies such as tissue clearing have expanded the field of observation available for fluorescence labeling; however, the area of correlative observation available for electron microscopy (EM) remains restricted. In this study, we developed a large-area CLEM imaging procedure to show specific molecular localization in large-scale EM sections of mouse and marmoset brain. Target molecules were labeled with antibodies and sequentially visualized in cryostat sections using fluorescence and gold particles. Fluorescence images were obtained by light microscopy immediately after antibody staining. Immunostained sections were postfixed for EM, and silver-enhanced sections were dehydrated in a graded ethanol series and embedded in resin. Ultrathin sections for EM were prepared from fully polymerized resin blocks, collected on silicon wafers, and observed by multibeam scanning electron microscopy (SEM). Multibeam SEM has made rapid, large-area observation at high resolution possible, paving the way for the analysis of detailed structures using the CLEM approach. Here, we describe detailed methods for large-area CLEM in various tissues of both rodents and primates.

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Noninvasive technique to evaluate the muscle fiber characteristics using q-space imaging

Hata J., Nakashima D., Tsuji O., Fujiyoshi K., Yasutake K., Sera Y., Komaki Y., Hikishima K., Nagura T., Matsumoto M., Okano H., Nakamura M.

PLoS ONE （PLoS ONE） 14 （ 4 ） 2019年04月

© 2019 Hata et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Background Skeletal muscles include fast and slow muscle fibers. The tibialis anterior muscle (TA) is mainly composed of fast muscle fibers, whereas the soleus muscle (SOL) is mainly composed of slow muscle fibers. However, a noninvasive approach for appropriately investigating the characteristics of muscles is not available. Monitoring of skeletal muscle characteristics can help in the evaluation of the effects of strength training and diseases on skeletal muscles. Purpose The present study aimed to determine whether q-space imaging can distinguish between TA and SOL in in vivo mice. Methods In vivo magnetic resonance imaging of the right calves of mice (n = 8) was performed using a 7-Tesla magnetic resonance imaging system with a cryogenic probe. TA and SOL were assessed. q-space imaging was performed with a field of view of 10 mm × 10 mm, matrix of 48 × 48, and section thickness of 1000 μm. There were ten b-values ranging from 0 to 4244 s/mm 2 , and each b-value had diffusion encoding in three directions. Magnetic resonance imaging findings were compared with immunohistological findings. Results Full width at half maximum and Kurtosis maps of q-space imaging showed signal intensities consistent with immunohistological findings for both fast (myosin heavy chain II) and slow (myosin heavy chain I) muscle fibers. With regard to quantification, both full width at half maximum and Kurtosis could represent the immunohistological findings that the cell diameter of TA was larger than that of SOL (P < 0.01). Conclusion q-space imaging could clearly differentiate TA from SOL using differences in cell diameters. This technique is a promising method to noninvasively estimate the fiber type ratio in skeletal muscles, and it can be further developed as an indicator of muscle characteristics.

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Application of hepatocyte growth factor for acute spinal cord injury: The road from basic studies to human treatment

Kitamura K., Nagoshi N., Tsuji O., Matsumoto M., Okano H., Nakamura M.

International Journal of Molecular Sciences （International Journal of Molecular Sciences） 20 （ 5 ） 2019年03月

ISSN 16616596

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. Hepatocyte growth factor (HGF) was first identified as a potent mitogen for mature hepatocytes, and has also gained attention as a strong neurotrophic factor in the central nervous system. We found that during the acute phase of spinal cord injury (SCI) in rats, c-Met, the specific receptor for HGF, increases sharply, while the endogenous HGF up-regulation is relatively weak. Introducing exogenous HGF into the spinal cord by injecting an HGF-expressing viral vector significantly increased the neuron and oligodendrocyte survival, angiogenesis, and axonal regeneration, to reduce the area of damage and to promote functional recovery in rats after SCI. Other recent studies in rodents have shown that exogenously administered HGF during the acute phase of SCI reduces astrocyte activation to decrease glial scar formation, and exerts anti-inflammatory effects to reduce leukocyte infiltration. We also reported that the intrathecal infusion of recombinant human HGF (intrathecal rhHGF) improves neurological hand function after cervical contusive SCI in the common marmoset, a non-human primate. Based on these collective results, we conducted a phase I/II clinical trial of intrathecal rhHGF for patients with acute cervical SCI who showed a modified Frankel grade of A/B1/B2 72 h after injury onset, from June 2014 to May 2018.

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Selective Ablation of Tumorigenic Cells Following Human Induced Pluripotent Stem Cell-Derived Neural Stem/Progenitor Cell Transplantation in Spinal Cord Injury

Kojima K., Miyoshi H., Nagoshi N., Kohyama J., Itakura G., Kawabata S., Ozaki M., Iida T., Sugai K., Ito S., Fukuzawa R., Yasutake K., Renault-Mihara F., Shibata S., Matsumoto M., Nakamura M., Okano H.

Stem Cells Translational Medicine （Stem Cells Translational Medicine） 8 （ 3 ） 260 - 270 2019年03月

ISSN 21576564

© 2018 The Authors Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press Tumorigenesis is an important problem that needs to be addressed in the field of human stem/progenitor cell transplantation for the treatment of subacute spinal cord injury (SCI). When certain “tumorigenic” cell lines are transplanted into the spinal cord of SCI mice model, there is initial improvement of motor function, followed by abrupt deterioration secondary to the effect of tumor growth. A significant proportion of the transplanted cells remains undifferentiated after transplantation and is thought to increase the risk of tumorigenesis. In this study, using lentiviral vectors, we introduced the herpes simplex virus type 1 thymidine kinase (HSVtk) gene into a human induced pluripotent stem cell-derived neural stem/progenitor cell (hiPSC-NS/PC) line that is known to undergo tumorigenic transformation. Such approach enables selective ablation of the immature proliferating cells and thereby prevents subsequent tumor formation. In vitro, the HSVtk system successfully ablated the immature proliferative neural cells while preserving mature postmitotic neuronal cells. Similar results were observed in vivo following transplantation into the injured spinal cords of immune-deficient (nonobese diabetic–severe combined immune-deficient) mice. Ablation of the proliferating cells exerted a protective effect on the motor function which was regained after transplantation, simultaneously defending the spinal cord from the harmful tumor growth. These results suggest a potentially promising role of suicide genes in opposing tumorigenesis during stem cell therapy. This system allows both preventing and treating tumorigenesis following hiPSC-NS/PC transplantation without sacrificing the improved motor function. Stem Cells Translational Medicine 2019;8:260&270.

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Induced Pluripotent Stem Cells Reprogrammed with Three Inhibitors Show Accelerated Differentiation Potentials with High Levels of 2-Cell Stage Marker Expression

Nishihara K., Shiga T., Nakamura E., Akiyama T., Sasaki T., Suzuki S., Ko M., Tada N., Okano H., Akamatsu W.

Stem Cell Reports （Stem Cell Reports） 12 （ 2 ） 305 - 318 2019年02月

ISSN 22136711

© 2019 The Authors Although pluripotent stem cells can generate various types of differentiated cells, it is unclear why lineage-committed stem/progenitor cells derived from pluripotent stem cells are decelerated and why the differentiation-resistant propensity of embryonic stem cell (ESC)/induced pluripotent stem cell (iPSC)-derived cells is predominant compared with the in vivo equivalents derived from embryonic/adult tissues. In this study, we demonstrated that iPSCs reprogrammed and maintained with three chemical inhibitors of the fibroblast growth factor 4-mitogen-activated protein kinase cascade and GSK3β (3i) could be differentiated into all three germ layers more efficiently than the iPSCs reprogrammed without the 3i chemicals, even though they were maintained with 3i chemicals once they were reprogrammed. Although the iPSCs reprogrammed with 3i had increased numbers of Zscan4-positive cells, the Zscan4-positive cells among iPSCs that were reprogrammed without 3i did not have an accelerated differentiation ability. These observations suggest that 3i exposure during the reprogramming period determines the accelerated differentiation/maturation potentials of iPSCs that are stably maintained at the distinct state.

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Increased cytotoxicity of herpes simplex virus thymidine kinase expression in human induced pluripotent stem cells

Iwasawa C., Tamura R., Sugiura Y., Suzuki S., Kuzumaki N., Narita M., Suematsu M., Nakamura M., Yoshida K., Toda M., Okano H., Miyoshi H.

International Journal of Molecular Sciences （International Journal of Molecular Sciences） 20 （ 4 ） 2019年02月

ISSN 16616596

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. Human induced pluripotent stem cells (iPSCs) hold enormous promise for regenerative medicine. The major safety concern is the tumorigenicity of transplanted cells derived from iPSCs. A potential solution would be to introduce a suicide gene into iPSCs as a safety switch. The herpes simplex virus type 1 thymidine kinase (HSV-TK) gene, in combination with ganciclovir, is the most widely used enzyme/prodrug suicide system from basic research to clinical applications. In the present study, we attempted to establish human iPSCs that stably expressed HSV-TK with either lentiviral vectors or CRISPR/Cas9-mediated genome editing. However, this task was difficult to achieve, because high-level and/or constitutive expression of HSV-TK resulted in the induction of cell death or silencing of HSV-TK expression. A nucleotide metabolism analysis suggested that excessive accumulation of thymidine triphosphate, caused by HSV-TK expression, resulted in an imbalance in the dNTP pools. This unbalanced state led to DNA synthesis inhibition and cell death in a process similar to a “thymidine block”, but more severe. We also demonstrated that the Tet-inducible system was a feasible solution for overcoming the cytotoxicity of HSV-TK expression. Our results provided a warning against using the HSV-TK gene in human iPSCs, particularly in clinical applications.

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Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Komatsu M., Kaneko T., Okano H., Ichinohe N.

Journal of visualized experiments : JoVE （Journal of visualized experiments : JoVE）（ 144 ） 2019年02月

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

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A high-throughput neurohistological pipeline for brain-wide mesoscale connectivity mapping of the common marmoset

Lin M., Takahashi Y., Huo B., Hanada M., Nagashima J., Hata J., Tolpygo A., Ram K., Lee B., Miller M., Rosa M., Sasaki E., Iriki A., Okano H., Mitra P.

eLife （eLife） 8 2019年02月

© Lin et al. Understanding the connectivity architecture of entire vertebrate brains is a fundamental but difficult task. Here we present an integrated neuro-histological pipeline as well as a grid-based tracer injection strategy for systematic mesoscale connectivity mapping in the common marmoset (Callithrix jacchus). Individual brains are sectioned into ~1700 20 mm sections using the tape transfer technique, permitting high quality 3D reconstruction of a series of histochemical stains (Nissl, myelin) interleaved with tracer labeled sections. Systematic in-vivo MRI of the individual animals facilitates injection placement into reference-atlas defined anatomical compartments. Further, by combining the resulting 3D volumes, containing informative cytoarchitectonic markers, with in-vivo and ex-vivo MRI, and using an integrated computational pipeline, we are able to accurately map individual brains into a common reference atlas despite the significant individual variation. This approach will facilitate the systematic assembly of a mesoscale connectivity matrix together with unprecedented 3D reconstructions of brain-wide projection patterns in a primate brain.

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Comparative principles for next-generation neuroscience

Miller C., Hale M., Okano H., Okabe S., Mitra P.

Frontiers in Behavioral Neuroscience （Frontiers in Behavioral Neuroscience） 13 2019年01月

ISSN 16625153

© 2019 Miller, Hale, Okano, Okabe and Mitra. Neuroscience is enjoying a renaissance of discovery due in large part to the implementation of next-generation molecular technologies. The advent of genetically encoded tools has complemented existing methods and provided researchers the opportunity to examine the nervous system with unprecedented precision and to reveal facets of neural function at multiple scales. The weight of these discoveries, however, has been technique-driven from a small number of species amenable to the most advanced gene-editing technologies. To deepen interpretation and build on these breakthroughs, an understanding of nervous system evolution and diversity are critical. Evolutionary change integrates advantageous variants of features into lineages, but is also constrained by pre-existing organization and function. Ultimately, each species’ neural architecture comprises both properties that are species-specific and those that are retained and shared. Understanding the evolutionary history of a nervous system provides interpretive power when examining relationships between brain structure and function. The exceptional diversity of nervous systems and their unique or unusual features can also be leveraged to advance research by providing opportunities to ask new questions and interpret findings that are not accessible in individual species. As new genetic and molecular technologies are added to the experimental toolkits utilized in diverse taxa, the field is at a key juncture to revisit the significance of evolutionary and comparative approaches for next-generation neuroscience as a foundational framework for understanding fundamental principles of neural function.

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Reduced expression of somatostatin in GABAergic interneurons derived from induced pluripotent stem cells of patients with parkin mutations

Iwasawa C., Kuzumaki N., Suda Y., Kagawa R., Oka Y., Hattori N., Okano H., Narita M.

Molecular Brain （Molecular Brain） 12 （ 1 ） 2019年01月

© 2019 The Author(s). Parkinson's disease (PD) is associated with both motor and non-motor symptoms, including constipation, sensory neuropathy, depression, dementia and sleep disorder. Somatostatin (SST) is considered to be a modulator of GABAergic inhibitory transmission, and its levels are reduced in cerebrospinal fluid of PD patients. In the present study, we evaluated the changes in the expression of SST in GABAergic neurons derived from induced pluripotent stem cells (iPSCs) of PD patients. Neural cells were co-treated with the Wnt antagonist IWP-2 and Shh during neurosphere formation to induce GABA-positive forebrain interneurons. Quantitative analyses showed no significant differences, but slight decreases, in the potency of differentiation into GABAergic neurons derived from iPSCs between healthy control and patients with PARK2 mutations, who have been classified as a type of early-onset familial PD due to mutations in the parkin gene. Under this condition, the mRNA level of SST in GABAergic interneurons derived from iPSCs of PARK2-specific PD patients significantly decreased as neural maturation progressed. We also found that SST-positive GABAergic neurons were clearly reduced in GABAergic neurons derived from iPSCs of patients with PARK2 mutations. These findings suggest that the reduction in the expression level of SST in GABAergic interneurons of PD may, at least partly, lead to complex PD-induced symptoms.

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Unidirectional monosynaptic connections from auditory areas to the primary visual cortex in the marmoset monkey

Majka P., Rosa M., Bai S., Chan J., Huo B., Jermakow N., Lin M., Takahashi Y., Wolkowicz I., Worthy K., Rajan R., Reser D., Wójcik D., Okano H., Mitra P.

Brain Structure and Function （Brain Structure and Function） 224 （ 1 ） 111 - 131 2019年01月

ISSN 18632653

© 2018, The Author(s). Until the late twentieth century, it was believed that different sensory modalities were processed by largely independent pathways in the primate cortex, with cross-modal integration only occurring in specialized polysensory areas. This model was challenged by the finding that the peripheral representation of the primary visual cortex (V1) receives monosynaptic connections from areas of the auditory cortex in the macaque. However, auditory projections to V1 have not been reported in other primates. We investigated the existence of direct interconnections between V1 and auditory areas in the marmoset, a New World monkey. Labelled neurons in auditory cortex were observed following 4 out of 10 retrograde tracer injections involving V1. These projections to V1 originated in the caudal subdivisions of auditory cortex (primary auditory cortex, caudal belt and parabelt areas), and targeted parts of V1 that represent parafoveal and peripheral vision. Injections near the representation of the vertical meridian of the visual field labelled few or no cells in auditory cortex. We also placed 8 retrograde tracer injections involving core, belt and parabelt auditory areas, none of which revealed direct projections from V1. These results confirm the existence of a direct, nonreciprocal projection from auditory areas to V1 in a different primate species, which has evolved separately from the macaque for over 30 million years. The essential similarity of these observations between marmoset and macaque indicate that early-stage audiovisual integration is a shared characteristic of primate sensory processing.

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Concise Review: Laying the Groundwork for a First-In-Human Study of an Induced Pluripotent Stem Cell-Based Intervention for Spinal Cord Injury

Tsuji O., Sugai K., Yamaguchi R., Tashiro S., Nagoshi N., Kohyama J., Iida T., Ohkubo T., Itakura G., Isoda M., Shinozaki M., Fujiyoshi K., Kanemura Y., Yamanaka S., Nakamura M., Okano H.

Stem Cells （Stem Cells） 37 （ 1 ） 6 - 13 2019年01月

ISSN 10665099

© 2018 The Authors Stem Cells published by Wiley Periodicals, Inc. on behalf of AlphaMed Press There have been numerous attempts to develop stem cell transplantation approaches to promote the regeneration of spinal cord injury (SCI). Our multicenter team is currently planning to launch a first-in-human clinical study of an induced pluripotent stem cell (iPSC)-based cell transplant intervention for subacute SCI. This trial was conducted as class I regenerative medicine protocol as provided for under Japan's Act on the Safety of Regenerative Medicine, using neural stem/progenitor cells derived from a clinical-grade, integration-free human “iPSC stock” generated by the Kyoto University Center for iPS Cell Research and Application. In the present article, we describe how we are preparing to initiate this clinical study, including addressing the issues of safety and tumorigenesis as well as practical problems that must be overcome to enable the development of therapeutic interventions for patients with chronic SCI. Stem Cells 2019;37:6–13.

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PAT - Probabilistic axon tracking for densely labeled neurons in large 3-D micrographs

Skibbe H., Reisert M., Nakae K., Watakabe A., Hata J., Mizukami H., Okano H., Yamamori T., Ishii S.

IEEE Transactions on Medical Imaging （IEEE Transactions on Medical Imaging） 38 （ 1 ） 69 - 78 2019年01月

ISSN 02780062

© 1982-2012 IEEE. A major goal of contemporary neuroscience research is to map the structural connectivity of mammalian brain using microscopy imaging data. In this context, the reconstruction of densely labeled axons from two-photon microscopy images is a challenging and important task. The visually overlapping, crossing, and often strongly distorted images of the axons allow many ambiguous interpretations to be made. We address the problem of tracking axons in densely labeled samples of neurons in large image data sets acquired from marmoset brains. Our high-resolution images were acquired using two-photon microscopy and they provided whole brain coverage, occupying terabytes of memory. Both the image distortions and the large data set size frequently make it impractical to apply present-day neuron tracing algorithms to such data due to the optimization of such algorithms to the precise tracing of either single or sparse sets of neurons. Thus, new tracking techniques are needed. We propose a probabilistic axon tracking algorithm (PAT). PAT tackles the tracking of axons in two steps: locally (L-PAT) and globally (G-PAT). L-PAT is a probabilistic tracking algorithm that can tackle distorted, cluttered images of densely labeled axons. L-PAT divides a large micrograph into smaller image stacks. It then processes each image stack independently before mapping the axons in each image to a sparse model of axon trajectories. G-PAT merges the sparse L-PAT models into a single global model of axon trajectories by minimizing a global objective function using a probabilistic optimization method. We demonstrate the superior performance of PAT over standard approaches on synthetic data. Furthermore, we successfully apply PAT to densely labeled axons in large images acquired from marmoset brains.

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Are We There Yet? How and When Specific Biotechnologies Will Improve Human Health

O'Day E., Hosta-Rigau L., Oyarzún D., Okano H., de Lorenzo V., von Kameke C., Alsafar H., Cao C., Chen G., Ji W., Roberts R., Ronaghi M., Yeung K., Zhang F., Lee S.

Biotechnology Journal （Biotechnology Journal） 14 （ 1 ） 2019年01月

ISSN 18606768

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Tau isoform expression and phosphorylation in marmoset brains

Sharma G., Huo A., Kimura T., Shiozawa S., Kobayashi R., Sahara N., Ishibashi M., Ishigaki S., Saito T., Ando K., Murayama S., Hasegawa M., Sobue G., Okano H., Hisanaga S.

Journal of Biological Chemistry （Journal of Biological Chemistry） 294 （ 30 ） 11433 - 11444 2019年

ISSN 00219258

© 2019 Sharma et al. Tau is a microtubule-associated protein expressed in neuronal axons. Hyperphosphorylated tau is a major component of neurofibrillary tangles, a pathological hallmark of Alzheimer’s disease (AD). Hyperphosphorylated tau aggregates are also found in many neurodegenerative diseases, collectively referred to as “tauopathies,” and tau mutations are associated with familial frontotemporal lobar degeneration (FTLD). Previous studies have generated transgenic mice with mutant tau as tauopathy models, but nonhuman primates, which are more similar to humans, may be a better model to study tauopathies. For example, the common marmoset is poised as a nonhuman primate model for investigating the etiology of age-related neurodegenerative diseases. However, no biochemical studies of tau have been conducted in marmoset brains. Here, we investigated several important aspects of tau, including expression of different tau isoforms and its phosphorylation status, in the marmoset brain. We found that marmoset tau does not possess the “primate-unique motif” in its N-terminal domain. We also discovered that the tau isoform expression pattern in marmosets is more similar to that of mice than that of humans, with adult marmoset brains expressing only four-repeat tau isoforms as in adult mice but unlike in adult human brains. Of note, tau in brains of marmoset newborns was phosphorylated at several sites associated with AD pathology. However, in adult marmoset brains, much of this phosphorylation was lost, except for Ser-202 and Ser-404 phosphorylation. These results reveal key features of tau expression and phosphorylation in the marmoset brain, a potentially useful nonhuman primate model of neurodegenerative diseases.

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Relation of koniocellular layers of dorsal lateral geniculate to inferior pulvinar nuclei in common marmosets

Huo B., Zeater N., Lin M., Takahashi Y., Hanada M., Nagashima J., Lee B., Hata J., Zaheer A., Grünert U., Miller M., Rosa M., Okano H., Martin P., Mitra P.

European Journal of Neuroscience （European Journal of Neuroscience） 2019年

ISSN 0953816X

© 2019 Federation of European Neuroscience Societies and John Wiley & Sons Ltd Traditionally, the dorsal lateral geniculate nucleus (LGN) and the inferior pulvinar (IPul) nucleus are considered as anatomically and functionally distinct thalamic nuclei. However, in several primate species it has also been established that the koniocellular (K) layers of LGN and parts of the IPul have a shared pattern of immunoreactivity for the calcium-binding protein calbindin. These calbindin-rich cells constitute a thalamic matrix system which is implicated in thalamocortical synchronisation. Further, the K layers and IPul are both involved in visual processing and have similar connections with retina and superior colliculus. Here, we confirmed the continuity between calbindin-rich cells in LGN K layers and the central lateral division of IPul (IPulCL) in marmoset monkeys. By employing a high-throughput neuronal tracing method, we found that both the K layers and IPulCL form comparable patterns of connections with striate and extrastriate cortices; these connections are largely different to those of the parvocellular and magnocellular laminae of LGN. Retrograde tracer-labelled cells and anterograde tracer-labelled axon terminals merged seamlessly from IPulCL into LGN K layers. These results support continuity between LGN K layers and IPulCL, providing an anatomical basis for functional congruity of this region of the dorsal thalamic matrix and calling into question the traditional segregation between LGN and the inferior pulvinar nucleus.

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In Utero Amniotic Fluid Stem Cell Therapy Protects Against Myelomeningocele via Spinal Cord Coverage and Hepatocyte Growth Factor Secretion

Abe Y., Ochiai D., Masuda H., Sato Y., Otani T., Fukutake M., Ikenoue S., Miyakoshi K., Okano H., Tanaka M.

Stem Cells Translational Medicine （Stem Cells Translational Medicine） 2019年

ISSN 21576564

© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press Despite the poor prognosis associated with myelomeningocele (MMC), the options for prenatal treatments are still limited. Recently, fetal cellular therapy has become a new option for treating birth defects, although the therapeutic effects and mechanisms associated with such treatments remain unclear. The use of human amniotic fluid stem cells (hAFSCs) is ideal with respect to immunoreactivity and cell propagation. The prenatal diagnosis of MMC during early stages of pregnancy could allow for the ex vivo proliferation and modulation of autologous hAFSCs for use in utero stem cell therapy. Therefore, we investigated the therapeutic effects and mechanisms of hAFSCs-based treatment for fetal MMC. hAFSCs were isolated as CD117-positive cells from the amniotic fluid of 15- to 17-week pregnant women who underwent amniocentesis for prenatal diagnosis and consented to this study. Rat dams were exposed to retinoic acid to induce fetal MMC and were subsequently injected with hAFSCs in each amniotic cavity. We measured the exposed area of the spinal cord and hepatocyte growth factor (HGF) levels at the lesion. The exposed spinal area of the hAFSC-treated group was significantly smaller than that of the control group. Immunohistochemical analysis demonstrated a reduction in neuronal damage such as neurodegeneration and astrogliosis in the hAFSC-treated group. Additionally, in lesions of the hAFSC-treated group, HGF expression was upregulated and HGF-positive hAFSCs were identified, suggesting that these cells migrated to the lesion and secreted HGF to suppress neuronal damage and induce neurogenesis. Therefore, in utero hAFSC therapy could become a novel strategy for fetal MMC. Stem Cells Translational Medicine 2019.

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Comparison of inner ear drug availability of combined treatment with systemic or local drug injections alone

Li Y., Kanzaki S., Shibata S., Nakamura M., Ozaki M., Okano H., Ogawa K.

Neuroscience Research （Neuroscience Research） 2019年

ISSN 01680102

© 2019 Combination of systemic and local drug therapy has been proved more effective and safer for idiopathic sudden sensorineural hearing loss by some clinical trials, and there are few laboratory researches on its pharmacokinetic behaviors in the inner ear. In the present study, we use a new in vivo imaging system to compare the pharmacokinetics of combined therapy (CT), with intravenous (IV) or transtympanic (TT) injection alone in both ears of transgenic GFAP-Luc mice. Biological half-life, total photon counts and the area under the curve (AUC) value significantly increased after CT. However, adding IV to TT injection cannot strengthen the peak photon of the drug in the inner ear. In addition, when D-luciferin is injected to the left ear the volume of total photon count and AUC value of CT-left ear are larger than the combined volume of TT-left ear and IV-left ear, suggesting a synergistic effect, and those of CT-right ear are almost equal to the summation of those of IV-right group and TT-right group, suggesting no amplifying effect on the risk of systemic side effect. This study showed that CT could deliver more drugs into the inner ear, and brought a longer therapeutic window, and were more effective than intravenous or transtympanic injection alone in the pharmacokinetics.

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A versatile toolbox for knock-in gene targeting based on the Multisite Gateway technology

Yoshimatsu S., Sone T., Nakajima M., Sato T., Okochi R., Ishikawa M., Nakamura M., Sasaki E., Shiozawa S., Okano H.

PLoS ONE （PLoS ONE） 14 （ 8 ） 2019年

© 2019 Yoshimatsu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Knock-in (KI) gene targeting can be employed for a wide range of applications in stem cell research. However, vectors for KI require multiple complicated processes for construction, including multiple times of digestion/ligation steps and extensive restriction mapping, which has imposed limitations for the robust applicability of KI gene targeting. To circumvent this issue, here we introduce versatile and systematic methods for generating KI vectors by molecular cloning. In this approach, we employed the Multisite Gateway technology, an efficient in vitro DNA recombination system using proprietary sequences and enzymes. KI vector construction exploiting these methods requires only efficient steps, such as PCR and recombination, enabling robust KI gene targeting. We show that combinatorial usage of the KI vectors generated using this method and site-specific nucleases enabled the precise integration of fluorescent protein genes in multiple loci of human and common marmoset (marmoset; Callithrix jacchus) pluripotent stem cells. The methods described here will facilitate the usage of KI technology and ultimately help to accelerate stem cell research.

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Treatment with a Gamma-Secretase Inhibitor Promotes Functional Recovery in Human iPSC- Derived Transplants for Chronic Spinal Cord Injury

Okubo T., Nagoshi N., Kohyama J., Tsuji O., Shinozaki M., Shibata S., Kase Y., Matsumoto M., Nakamura M., Okano H.

Stem Cell Reports （Stem Cell Reports） 11 （ 6 ） 1416 - 1432 2018年12月

ISSN 22136711

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Engraftment of Human Pluripotent Stem Cell-derived Progenitors in the Inner Ear of Prenatal Mice

Takeda H., Hosoya M., Fujioka M., Saegusa C., Saeki T., Miwa T., Okano H., Minoda R.

Scientific Reports （Scientific Reports） 8 （ 1 ） 2018年12月

© 2018 The Author(s). There is, at present, no curative treatment for genetic hearing loss. We have previously reported that transuterine gene transfer of wild type CONNEXIN30 (CX30) genes into otocysts in CX30-deleted mice could restore hearing. Cell transplantation therapy might be another therapeutic option, although it is still unknown whether stem cell-derived progenitor cells could migrate into mouse otocysts. Here, we show successful cell transplantation of progenitors of outer sulcus cell-like cells derived from human-derived induced pluripotent stem cells into mouse otocysts on embryonic day 11.5. The delivered cells engrafted more frequently in the non-sensory region in the inner ear of CX30-deleted mice than in wild type mice and survived for up to 1 week after transplantation. Some of the engrafted cells expressed CX30 proteins in the non-sensory region. This is the first report that demonstrates successful engraftment of exogenous cells in prenatal developing otocysts in mice. Future studies using this mouse otocystic injection model in vivo will provide further clues for developing treatment modalities for congenital hearing loss in humans.

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Assessing cortical plasticity after spinal cord injury by using resting-state functional magnetic resonance imaging in awake adult mice

Matsubayashi K., Nagoshi N., Komaki Y., Kojima K., Shinozaki M., Tsuji O., Iwanami A., Ishihara R., Takata N., Matsumoto M., Mimura M., Okano H., Nakamura M.

Scientific Reports （Scientific Reports） 8 （ 1 ） 2018年12月

© 2018, The Author(s). Neural connectivity has recently been shown to be altered after spinal cord injury (SCI) not only in the spinal cord but also in the brain. However, to date, no studies have analyzed the functional alterations after SCI in various areas of the cerebral cortex over time. To examine the plasticity of the neural connectivity in the brain after SCI, we performed resting-state functional magnetic resonance imaging (rs-fMRI) in awake adult mice pre- and post-SCI. After a complete thoracic SCI, the functional connectivity between the primary motor (MOp) and primary sensory (SSp) areas was significantly decreased during the chronic phase. In contrast, the connectivity between the MOp and motivation area was increased. Thus, impairments in sensory and motor connections after SCI led to a time-dependent compensatory upregulation of “motor functional motivation”. Moreover, the functional connectivity between the SSp and pain-related areas, such as the caudoputamen (CP) and the anterior cingulate area (ACA), was strengthened during the chronic phase, thus suggesting that rs-fMRI can indicate the presence of neuropathic pain after SCI. Therefore, rs-fMRI is a useful tool for revealing the pathological changes that occur in the brain after SCI.

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Single-cell trajectory analysis of human homogenous neurons carrying a rare RELN variant

Arioka Y., Shishido E., Kubo H., Kushima I., Yoshimi A., Kimura H., Ishizuka K., Aleksic B., Maeda T., Ishikawa M., Kuzumaki N., Okano H., Mori D., Ozaki N.

Translational Psychiatry （Translational Psychiatry） 8 （ 1 ） 2018年12月

© 2018 The Author(s). Reelin is a protein encoded by the RELN gene that controls neuronal migration in the developing brain. Human genetic studies suggest that rare RELN variants confer susceptibility to mental disorders such as schizophrenia. However, it remains unknown what effects rare RELN variants have on human neuronal cells. To this end, the analysis of human neuronal dynamics at the single-cell level is necessary. In this study, we generated human-induced pluripotent stem cells carrying a rare RELN variant (RELN-del) using targeted genome editing; cells were further differentiated into highly homogeneous dopaminergic neurons. Our results indicated that RELN-del triggered an impaired reelin signal and decreased the expression levels of genes relevant for cell movement in human neurons. Single-cell trajectory analysis revealed that control neurons possessed directional migration even in vitro, while RELN-del neurons demonstrated a wandering type of migration. We further confirmed these phenotypes in neurons derived from a patient carrying the congenital RELN-del. To our knowledge, this is the first report of the biological significance of a rare RELN variant in human neurons based on individual neuron dynamics. Collectively, our approach should be useful for studying reelin function and evaluating mental disorder susceptibility, focusing on individual human neuronal migration.

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The pathogenesis linked to coenzyme Q10 insufficiency in iPSC-derived neurons from patients with multiple-system atrophy

Nakamoto F., Okamoto S., Mitsui J., Sone T., Ishikawa M., Yamamoto Y., Kanegae Y., Nakatake Y., Imaizumi K., Ishiura H., Tsuji S., Okano H.

Scientific Reports （Scientific Reports） 8 （ 1 ） 2018年12月

© 2018, The Author(s). Multiple-system atrophy (MSA) is a neurodegenerative disease characterized by autonomic failure with various combinations of parkinsonism, cerebellar ataxia, and pyramidal dysfunction. We previously reported that functionally impaired variants of COQ2, which encodes an essential enzyme in the biosynthetic pathway of coenzyme Q10, are associated with MSA. Here, we report functional deficiencies in mitochondrial respiration and the antioxidative system in induced pluripotent stem cell (iPSC)-derived neurons from an MSA patient with compound heterozygous COQ2 mutations. The functional deficiencies were rescued by site-specific CRISPR/Cas9-mediated gene corrections. We also report an increase in apoptosis of iPSC-derived neurons from MSA patients. Coenzyme Q10 reduced apoptosis of neurons from the MSA patient with compound heterozygous COQ2 mutations. Our results reveal that cellular dysfunctions attributable to decreased coenzyme Q10 levels are related to neuronal death in MSA, particularly in patients with COQ2 variants, and may contribute to the development of therapy using coenzyme Q10 supplementation.

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Stem cells purified from human induced pluripotent stem cell-derived neural crest-like cells promote peripheral nerve regeneration

Kimura H., Ouchi T., Shibata S., Amemiya T., Nagoshi N., Nakagawa T., Matsumoto M., Okano H., Nakamura M., Sato K.

Scientific Reports （Scientific Reports） 8 （ 1 ） 2018年12月

© 2018 The Author(s). Strategies for therapeutic cell transplantation have been assessed for use in the treatment of massive peripheral nerve defects. To support safe and efficient cell transplantation, we have focused on the purification of cells using cell surface markers. Our group previously reported low-Affinity nerve growth factor receptor (LNGFR)-and thymocyte antigen-1 (THY-1)-positive neural crest-like cells (LT-NCLCs), generated from human induced pluripotent stem cells (hiPSCs). In the present study, we investigated the efficacy of transplantation of hiPSC-derived LT-NCLCs in a murine massive peripheral nerve defect model. Animals with a sciatic nerve defect were treated with a bridging silicone tube prefilled with LT-NCLCs or medium in the transplantation (TP) and negative control (NC) groups, respectively. The grafted LT-NCLCs survived and enhanced myelination and angiogenesis, as compared to the NC group. Behavioral analysis indicated that motor functional recovery in the TP group was superior to that in the NC group, and similar to that in the autograft (Auto) group. LT-NCLCs promoted axonal regrowth and remyelination by Schwann cells. Transplantation of LT-NCLCs is a promising approach for nerve regeneration treatment of massive peripheral nerve defects.

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T-type Calcium Channels Determine the Vulnerability of Dopaminergic Neurons to Mitochondrial Stress in Familial Parkinson Disease

Tabata Y., Imaizumi Y., Sugawara M., Andoh-Noda T., Banno S., Chai M., Sone T., Yamazaki K., Ito M., Tsukahara K., Saya H., Hattori N., Kohyama J., Okano H.

Stem Cell Reports （Stem Cell Reports） 11 （ 5 ） 1171 - 1184 2018年11月

ISSN 22136711

© 2018 The Author(s) Parkinson disease (PD) is a progressive neurological disease caused by selective degeneration of dopaminergic (DA) neurons in the substantia nigra. Although most cases of PD are sporadic cases, familial PD provides a versatile research model for basic mechanistic insights into the pathogenesis of PD. In this study, we generated DA neurons from PARK2 patient-specific, isogenic PARK2 null and PARK6 patient-specific induced pluripotent stem cells and found that these neurons exhibited more apoptosis and greater susceptibility to rotenone-induced mitochondrial stress. From phenotypic screening with an FDA-approved drug library, one voltage-gated calcium channel antagonist, benidipine, was found to suppress rotenone-induced apoptosis. Furthermore, we demonstrated the dysregulation of calcium homeostasis and increased susceptibility to rotenone-induced stress in PD, which is prevented by T-type calcium channel knockdown or antagonists. These findings suggest that calcium homeostasis in DA neurons might be a useful target for developing new drugs for PD patients. Our study demonstrate the dysregulation of calcium homeostasis and increased susceptibility to rotenone-induced stress in PD patient-derived DA neurons, which are further prevented by T-type calcium channel antagonists. These findings suggest that calcium homeostasis in DA neurons would be a useful target for developing new drugs for PD patients.

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The Amelioration of Pain-Related Behavior in Mice with Chronic Spinal Cord Injury Treated with Neural Stem/Progenitor Cell Transplantation Combined with Treadmill Training

Tashiro S., Nishimura S., Shinozaki M., Takano M., Konomi T., Tsuji O., Nagoshi N., Toyama Y., Liu M., Okano H., Nakamura M.

Journal of Neurotrauma （Journal of Neurotrauma） 35 （ 21 ） 2561 - 2571 2018年11月

ISSN 08977151

© Copyright 2018, Mary Ann Liebert, Inc., publishers 2018. Progress in regenerative medicine is realizing the possibility of neural regeneration and functional recovery in spinal cord injury (SCI). Recently, rehabilitation has attracted much attention with respect to the synergistic promotion of functional recovery in combination with neural stem/progenitor cell (NS/PC) transplantation, even in the chronic refractory phase of SCI. Nevertheless, sensory disturbance is one of the most prominent sequelae, even though the effects of combination or single therapies have been investigated mostly in the context of motor recovery. To determine how combination therapy with treadmill training (TMT) and NS/PC transplantation affects the manifestation of thermal allodynia and tactile hyperalgesia in chronic phase SCI, four groups of SCI mice were used to assess pain-related behavior and histological changes: combined transplantation and TMT therapy, transplantation only, TMT only, and control groups. Thermal allodynia and coarse touch-pressure hyperalgesia exhibited significant recovery in the combined therapy group in comparison with controls, whereas there were no significant differences with fine touch-pressure hyperalgesia and motor function. Further investigation revealed fewer fibers remaining in the posterior funiculus, which contained the tracts associated with the two modalities showing less recovery; that is, touch-pressure hyperalgesia and motor function. A significant correlation was only observed between these two modalities. Although no remarkable histological recovery was found within the lesion epicenter, changes indicating amelioration of pain were observed in the lumbar enlargement of the combination therapy group. Our results suggest that amelioration of thermal allodynia and tactile hyperalgesia can be brought about by the additive effect of NS/PC transplantation and TMT. The degree of recovery seems dependent on the distribution of damage.

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The Japan Monkey Centre Primates Brain Imaging Repository for comparative neuroscience: an archive of digital records including records for endangered species

Sakai T., Hata J., Ohta H., Shintaku Y., Kimura N., Ogawa Y., Sogabe K., Mori S., Okano H., Hamada Y., Shibata S., Okano H., Oishi K.

Primates （Primates） 59 （ 6 ） 553 - 570 2018年11月

ISSN 00328332

© 2018, The Author(s). Advances in magnetic resonance imaging (MRI) and computational analysis technology have enabled comparisons among various primate brains in a three-dimensional electronic format. Results from comparative studies provide information about common features across primates and species-specific features of neuroanatomy. Investigation of various species of non-human primates is important for understanding such features, but the majority of comparative MRI studies have been based on experimental primates, such as common marmoset, macaques, and chimpanzee. A major obstacle has been the lack of a database that includes non-experimental primates’ brain MRIs. To facilitate scientific discoveries in the field of comparative neuroanatomy and brain evolution, we launched a collaborative project to develop an open-resource repository of non-human primate brain images obtained using ex vivo MRI. As an initial open resource, here we release a collection of structural MRI and diffusion tensor images obtained from 12 species: pygmy marmoset, owl monkey, white-fronted capuchin, crab-eating macaque, Japanese macaque, bonnet macaque, toque macaque, Sykes’ monkey, red-tailed monkey, Schmidt’s guenon, de Brazza’s guenon, and lar gibbon. Sixteen postmortem brain samples from the 12 species, stored in the Japan Monkey Centre (JMC), were scanned using a 9.4-T MRI scanner and made available through the JMC collaborative research program (http://www.j-monkey.jp/BIR/index_e.html). The expected significant contributions of the JMC Primates Brain Imaging Repository include (1) resources for comparative neuroscience research, (2) preservation of various primate brains, including those of endangered species, in a permanent digital form, (3) resources with higher resolution for identifying neuroanatomical features, compared to previous MRI atlases, (4) resources for optimizing methods of scanning large fixed brains, and (5) references for veterinary neuroradiology. User-initiated research projects beyond these contributions are also anticipated.

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Versatile Roles of LKB1 Kinase Signaling in Neural Development and Homeostasis

Kuwako K., Okano H.

Frontiers in Molecular Neuroscience （Frontiers in Molecular Neuroscience） 11 2018年10月

ISSN 16625099

© 2018 Kuwako and Okano. Kinase signaling pathways orchestrate a majority of cellular structures and functions across species. Liver kinase B1 (LKB1, also known as STK11 or Par-4) is a ubiquitously expressed master serine/threonine kinase that plays crucial roles in numerous cellular events, such as polarity control, proliferation, differentiation and energy homeostasis, in many types of cells by activating downstream kinases of the AMP-activated protein kinase (AMPK) subfamily members. In contrast to the accumulating evidence for LKB1 functions in nonneuronal tissues, its functions in the nervous system have been relatively less understood until recently. In the brain, LKB1 initially emerged as a principal regulator of axon/dendrite polarity in forebrain neurons. Thereafter, recent investigations have rapidly uncovered diverse and essential functions of LKB1 in the developing and mature nervous system, such as migration, neurite morphogenesis, myelination and the maintenance of neural integrity, demonstrating that LKB1 is also a multifunctional master kinase in the nervous system. In this review article, we summarize the expanding knowledge about the functional aspects of LKB1 signaling in neural development and homeostasis.

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Modeling sporadic ALS in iPSC-derived motor neurons identifies a potential therapeutic agent

Fujimori K., Ishikawa M., Otomo A., Atsuta N., Nakamura R., Akiyama T., Hadano S., Aoki M., Saya H., Sobue G., Okano H.

Nature Medicine （Nature Medicine） 24 （ 10 ） 1579 - 1589 2018年10月

ISSN 10788956

© 2018, The Author(s). Amyotrophic lateral sclerosis (ALS) is a heterogeneous motor neuron disease for which no effective treatment is available, despite decades of research into SOD1-mutant familial ALS (FALS). The majority of ALS patients have no familial history, making the modeling of sporadic ALS (SALS) essential to the development of ALS therapeutics. However, as mutations underlying ALS pathogenesis have not yet been identified, it remains difficult to establish useful models of SALS. Using induced pluripotent stem cell (iPSC) technology to generate stem and differentiated cells retaining the patients’ full genetic information, we have established a large number of in vitro cellular models of SALS. These models showed phenotypic differences in their pattern of neuronal degeneration, types of abnormal protein aggregates, cell death mechanisms, and onset and progression of these phenotypes in vitro among cases. We therefore developed a system for case clustering capable of subdividing these heterogeneous SALS models by their in vitro characteristics. We further evaluated multiple-phenotype rescue of these subclassified SALS models using agents selected from non-SOD1 FALS models, and identified ropinirole as a potential therapeutic candidate. Integration of the datasets acquired in this study permitted the visualization of molecular pathologies shared across a wide range of SALS models.

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Quantitative temporal changes in DTI values coupled with histological properties in cuprizone-induced demyelination and remyelination

Yano R., Hata J., Abe Y., Seki F., Yoshida K., Komaki Y., Okano H., Tanaka K.

Neurochemistry International （Neurochemistry International） 119 151 - 158 2018年10月

ISSN 01970186

© 2017 Elsevier Ltd Diffusion tensor imaging (DTI) is widely used to evaluate microstructural variations in brain tissue. In particular, fractional anisotropy (FA), reflecting the magnitude and orientation of anisotropic water diffusion, allows us to detect pathological events in white matter. An ex vivo DTI study coupled with histological assessment is an efficient strategy to evaluate the myelination process, i.e. demyelination and remyelination. The relationship between DTI values and myelin content or the individual cellular components such as oligodendrocytes, microglia, and astrocytes during both processes of demyelination and remyelination are not well-understood. To address this issue, we employed a cuprizone-inducible demyelination mouse model. Demyelination can be induced in this model during cuprizone exposure and termination of cuprizone exposure induces remyelination. We fed the mice cuprizone-containing chow for 4 weeks and then normal chow for an additional 4 weeks. The ex vivo DTI was performed to evaluate the white matter profiles observed by FA, mean diffusivity (MD), and radial diffusivity (RD) at both demyelinating and remyelinating time points, and then we evaluated histological properties at the same time points. The results indicated a gradual FA decrease during the cuprizone treatment (0, 2, 3, 4 weeks). A lower peak was seen at 1 week after the normal chow was resumed, with recovery to baseline at 2 and 4 weeks. MD and RD showed an opposing pattern to that of FA. These DTI values were positively or negatively correlated with myelin content regardless of the status of the white matter. The RD value was more sensitive to myelination status than FA and MD. We have clarified the temporal changes in the DTI values coupled with histological properties over both the demyelination and remyelination processes.

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Control of directionality of chromatin folding for the inter- and intra-domain contacts at the Tfap2c-Bmp7 locus

Tsujimura T., Takase O., Yoshikawa M., Sano E., Hayashi M., Takato T., Toyoda A., Okano H., Hishikawa K.

Epigenetics and Chromatin （Epigenetics and Chromatin） 11 （ 1 ） 2018年09月

© 2018 The Author(s). Background: Contact domains of chromatin serve as a fundamental unit to regulate action of enhancers for target genes. Looping between a pair of CCCTC-binding factor (CTCF)-binding sites in convergent orientations underlies the formation of contact domains, while those in divergent orientations establish domain boundaries. However, every CTCF site is not necessarily engaged in loop or boundary structures, leaving functions of CTCF in varied genomic contexts still elusive. The locus containing Tfap2c and Bmp7 encompasses two contact domains separated by a region between the two genes, termed transition zone (TZ), characterized by two arrays of CTCF sites in divergent configuration. In this study, we created deletion and inversion alleles of these and other regions across the locus and investigated how they impinge on the conformation. Results: Deletion of the whole two CTCF arrays with the CRISPR/Cas9 system resulted in impairment of blocking of chromatin contacts by the TZ, as assessed by the circular chromatin conformation capture assay (4C-seq). Deletion and inversion of either of the two arrays similarly, but less pronouncedly, led to reduction in the blocking activity. Thus, the divergent configuration provides the TZ with the strong boundary activity. Uniquely, we show the TZ harbors a 50-kb region within one of the two arrays that contacts broadly with the both flanking intervals, regardless of the presence or orientation of the other CTCF array. Further, we show the boundary CTCF array has little impact on intra-domain folding; instead, locally associating CTCF sites greatly affect it. Conclusions: Our results show that the TZ not only separates the two domains, but also bears a wide interval that shows isotropic behavior of chromatin folding, indicating a potentially complex nature of actual boundaries in the genome. We also show that CTCF-binding sites inside a domain greatly contribute to the intra-domain folding of chromatin. Thus, the study reveals diverse and context-dependent roles of CTCF in organizing chromatin conformation at different levels.

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Responses to pup vocalizations in subordinate naked mole-rats are induced by estradiol ingested through coprophagy of queen’s feces

Watarai A., Arai N., Miyawaki S., Okano H., Miura K., Mogi K., Kikusui T.

Proceedings of the National Academy of Sciences of the United States of America （Proceedings of the National Academy of Sciences of the United States of America） 115 （ 37 ） 9264 - 9269 2018年09月

ISSN 00278424

© 2018 National Academy of Sciences. All Rights Reserved. Naked mole-rats form eusocial colonies consisting of a single breeding female (the queen), several breeding males, and sexually immature adults (subordinates). Subordinates are cooperative and provide alloparental care by huddling and retrieving pups to the nest. However, the physiological mechanism(s) underlying alloparental behavior of nonbreeders remains undetermined. Here, we examined the response of subordinates to pup voice and the fecal estradiol concentrations of subordinates during the three reproductive periods of the queen, including gestation, postpartum, and nonlactating. Subordinate response to pup voice was observed only during the queen’s postpartum and was preceded by an incremental rise in subordinates’ fecal estradiol concentrations during the queen’s gestation period, which coincided with physiological changes in the queen. We hypothesized that the increased estradiol in the queen’s feces was disseminated to subordinates through coprophagy, which stimulated subordinates’ responses to pup vocalizations. To test this hypothesis, we fed subordinates either fecal pellets from pregnant queens or pellets from nonpregnant queens amended with estradiol for 9 days and examined their response to recorded pup voice. In both treatments, the subordinates exhibited a constant level of response to pup voice during the feeding period but became more responsive 4 days after the feeding period. Thus, we believe that we have identified a previously unknown system of communication in naked mole-rats, in which a hormone released by one individual controls the behavior of another individual and influences the level of responsiveness among subordinate adults to pup vocal signals, thereby contributing to the alloparental pup care by subordinates.

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The LKB1-SIK Pathway Controls Dendrite Self-Avoidance in Purkinje Cells

Kuwako K., Okano H.

Cell Reports （Cell Reports） 24 （ 11 ） 2808 - 2818.e4 2018年09月

© 2018 The Author(s) Strictly controlled dendrite patterning underlies precise neural connection. Dendrite self-avoidance is a crucial system preventing self-crossing and clumping of dendrites. Although many cell-surface molecules that regulate self-avoidance have been identified, the signaling pathway that orchestrates it remains poorly understood, particularly in mammals. Here, we demonstrate that the LKB1-SIK kinase pathway plays a pivotal role in the self-avoidance of Purkinje cell (PC) dendrites by ensuring dendritic localization of Robo2, a regulator of self-avoidance. LKB1 is activated in developing PCs, and PC-specific deletion of LKB1 severely disrupts the self-avoidance of PC dendrites without affecting gross morphology. SIK1 and SIK2, downstream kinases of LKB1, mediate LKB1-dependent dendrite self-avoidance. Furthermore, loss of LKB1 leads to significantly decreased Robo2 levels in the dendrite but not in the cell body. Finally, restoration of dendritic Robo2 level via overexpression largely rescues the self-avoidance defect in LKB1-deficient PCs. These findings reveal an LKB1-pathway-mediated developmental program that establishes dendrite self-avoidance. Kuwako and Okano show that the LKB1-SIK axis is essential for establishing dendrite self-avoidance in Purkinje cells by ensuring an appropriate level of dendritic Robo2. This study reveals a pivotal role for the LKB1 kinase pathway in the developmental program orchestrating the spatial configuration of dendrites in mammalian brain.

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LOTUS Inhibits Neuronal Apoptosis and Promotes Tract Regeneration in Contusive Spinal Cord Injury Model Mice

Ito S., Nagoshi N., Tsuji O., Shibata S., Shinozaki M., Kawabata S., Kojima K., Yasutake K., Hirokawa T., Matsumoto M., Takei K., Nakamura M., Okano H.

eNeuro （eNeuro） 5 （ 5 ） 2018年09月

Nogo receptor-1 (NgR1) signaling is involved in the limitation of axonal regeneration following spinal cord injury (SCI) through collapsing the growth cone and inhibiting neurite outgrowth. Lateral olfactory tract usher substance (LOTUS), a NgR antagonist, suppresses these pathological conditions. A previous report demonstrated the positive effects of LOTUS expression on motor function through raphespinal tract regeneration using pan-neuronally LOTUS-overexpressing transgenic mice. However, this report used a hemi-section model, which does not represent the majority of clinical SCI cases, and lacked a detailed histological analysis of other descending tracts. To determine the true therapeutic effects of LOTUS, we used a more clinically relevant contusive SCI model in female transgenic mice. Definitive tracing analyses revealed that LOTUS promoted the extensive regeneration of the reticulospinal tract across the lesion site and suppressed axonal dieback of corticospinal tract (CST). A significant increase in raphespinal tract fibers was seen from the subacute to the chronic phase after the injury, strongly suggesting that LOTUS promoted translesional elongation of this tract. Furthermore, histological analyses revealed that LOTUS had a neuroprotective effect on the injured spinal cord through suppressing cellular apoptosis during the acute phase. These neuroprotective and regenerative effects contributed to significant motor functional recovery and restoration of the motor evoked potential (MEP). Therefore, LOTUS application could prove beneficial in the treatment of SCI by promoting axonal regeneration of some descending fibers, reducing axonal dieback of CST fibers and encouraging motor function recovery.

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Optogenetic astrocyte activation evokes BOLD fMRI response with oxygen consumption without neuronal activity modulation

Takata N., Sugiura Y., Yoshida K., Koizumi M., Hiroshi N., Honda K., Yano R., Komaki Y., Matsui K., Suematsu M., Mimura M., Okano H., Tanaka K.

GLIA （GLIA） 66 （ 9 ） 2013 - 2023 2018年09月

ISSN 08941491

© 2018 Wiley Periodicals, Inc. Functional magnetic resonance imaging (fMRI) based on the blood oxygenation level-dependent (BOLD) signal has been used to infer sites of neuronal activation in the brain. A recent study demonstrated, however, unexpected BOLD signal generation without neuronal excitation, which led us to hypothesize the presence of another cellular source for BOLD signal generation. Collective assessment of optogenetic activation of astrocytes or neurons, fMRI in awake mice, electrophysiological measurements, and histochemical detection of neuronal activation, coherently suggested astrocytes as another cellular source. Unexpectedly, astrocyte-evoked BOLD signal accompanied oxygen consumption without modulation of neuronal activity. Imaging mass spectrometry of brain sections identified synthesis of acetyl-carnitine via oxidative glucose metabolism at the site of astrocyte-, but not neuron-evoked BOLD signal. Our data provide causal evidence that astrocytic activation alone is able to evoke BOLD signal response, which may lead to reconsideration of current interpretation of BOLD signal as a marker of neuronal activation.

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Calcium Transient Dynamics of Neural Ensembles in the Primary Motor Cortex of Naturally Behaving Monkeys

Kondo T., Saito R., Otaka M., Yoshino-Saito K., Yamanaka A., Yamamori T., Watakabe A., Mizukami H., Schnitzer M., Tanaka K., Ushiba J., Okano H.

Cell Reports （Cell Reports） 24 （ 8 ） 2191 - 2195.e4 2018年08月

© 2018 The Authors To understand brain circuits of cognitive behaviors under natural conditions, we developed techniques for imaging neuronal activities from large neuronal populations in the deep layer cortex of the naturally behaving common marmoset. Animals retrieved food pellets or climbed ladders as a miniature fluorescence microscope monitored hundreds of calcium indicator-expressing cortical neurons in the right primary motor cortex. This technique, which can be adapted to other brain regions, can deepen our understanding of brain circuits by facilitating longitudinal population analyses of neuronal representation associated with cognitive naturalistic behaviors and their pathophysiological processes. Kondo et al. demonstrate calcium imaging from neuronal populations using a miniature fluorescence microscope in naturally behaving non-human primates (NHPs), common marmosets. This technique marks an advance beyond methods that use fixed head positioning, which limits the study of complex, self-determined behaviors such as social interactions or fear and anxiety.

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Establishment of a human induced stem cell line (FUi002-A) from Dravet syndrome patient carrying heterozygous R1525X mutation in SCN1A gene

Tanaka Y., Higurashi N., Shirasu N., Yasunaga S., Moreira K., Okano H., Hirose S.

Stem Cell Research （Stem Cell Research） 31 11 - 15 2018年08月

ISSN 18735061

© 2018 The Authors De novo mutations in SCN1A are the most common cause of Dravet syndrome (DS), an infantile-onset epileptic encephalopathy. In this study, human induced pluripotent stem cell (hiPSC) line FUi002-A was generated from skin fibroblasts obtained from a clinically diagnosed 26-year-old male DS patient with the R1525X variant of the SCN1A gene. Skin fibroblasts were reprogrammed using OriP/EBNA-1 based episomal plasmids expressing reprogramming factors expressing OCT4, SOX2, KLF-4, L-MYC, LIN28, and p53 shRNA. The transgene-free FUi002-A showed pluripotency, three germ layer differentiation capacity in vitro, and a normal karyotype. The resulting hiPSCs were heterozygous for the mutation in the SCN1A gene.

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HER2 heterogeneity is associated with poor survival in HER2-positive breast cancer

Hosonaga M., Arima Y., Sampetrean O., Komura D., Koya I., Sasaki T., Sato E., Okano H., Kudoh J., Ishikawa S., Saya H., Ishikawa T.

International Journal of Molecular Sciences （International Journal of Molecular Sciences） 19 （ 8 ） 2018年08月

ISSN 16616596

© 2018 by the authors. Licensee MDPI, Basel, Switzerland. Intratumoral human epidermal growth factor receptor 2 (HER2) heterogeneity has been reported in 16–36% of HER2-positive breast cancer and its clinical impact is under discussion. We examined the biological effects of HER2-heterogeneity on mouse models and analyzed metastatic brains by RNA sequence analysis. A metastatic mouse model was developed using 231-Luc (triple negative cells) and 2 HER2-positive cell lines, namely, HER2-60 and HER2-90 which showed heterogeneous and monotonous HER2 expressions, respectively. Metastatic lesions developed in 3 weeks in all the mice injected with HER2-60 cells, and in 69% of the mice injected with HER2-90 and 87.5% of the mice injected with 231-Luc. The median survival days of mice injected with 231-Luc, HER2-60, and HER2-90 cells were 29 (n = 24), 24 (n = 22) and 30 (n = 13) days, respectively. RNA sequence analysis showed that CASP-1 and its related genes were significantly downregulated in metastatic brain tumors with HER2-60 cells. The low expression of caspase-1 could be a new prognostic biomarker for early relapse in HER2-positive breast cancer.

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Recurrent Spindle Cell Carcinoma Shows Features of Mesenchymal Stem Cells

Ouchi T., Morikawa S., Shibata S., Takahashi M., Yoshikawa M., Soma T., Miyashita H., Muraoka W., Kameyama K., Kawana H., Arima Y., Saya H., Okano H., Nakagawa T., Asoda S.

Journal of Dental Research （Journal of Dental Research） 97 （ 7 ） 779 - 786 2018年07月

ISSN 00220345

© 2018, International & American Associations for Dental Research. This study investigated a case of spindle cell carcinoma (SpCC) in tongue pathological lesions. The patient experienced a local recurrence and distant metastasis after surgical intervention. Although standard chemotherapy was administered, a granulomatous mass continued to develop. This aggressive growth led to survival of the tumor. Secondary debulking surgery was performed to improve the patient’s quality of life at the request of the patient. Using a tissue sample derived from the secondary debulking surgery, we performed an analysis of the tumor’s cell surface antigens, differentiation potential, metastatic ability, and inhibition potential by anticancer reagents. In vitro analysis revealed that the cell population grown under adherent culture conditions expressed the mesenchymal stem cell (MSC) markers CD73, CD90, and CD105. The cell line established from this SpCC contained colony-forming unit fibroblasts (CFU-Fs) and exhibited multipotent differentiation into several mesenchymal lineages, including bone, cartilage, and fat. The SpCC cells also displayed vigorous mobilization. These characteristics suggested that they had the differentiation potential of mesenchymal cells, especially MSCs, rather than that of epithelial cells. The surgical specimen analyzed in this study resisted the molecular target reagent cetuximab, which is an epidermal growth factor receptor inhibitor. This clinical insight revealed that chemotherapy-resistant SpCC cells have different characteristics compared to most other cancer cells, which are sensitive to cetuximab. Our cell death assay revealed that SpCC cell death was induced by the anticancer drug imatinib, which is known to inhibit protein tyrosine kinase activity of ABL, platelet-derived growth factor receptor α (PDGFRα), and KIT. Here, we report recurrent SpCC with characteristics of MSCs and potential for treatment with imatinib.

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Investigation of brain science and neurological/psychiatric disorders using genetically modified non-human primates

Okano H., Kishi N.

Current Opinion in Neurobiology （Current Opinion in Neurobiology） 50 1 - 6 2018年06月

ISSN 09594388

© 2017 Elsevier Ltd Although mice have been the most frequently used experimental animals in many research fields due to well-established gene manipulation techniques, recent evidence has revealed that rodent models do not always recapitulate pathophysiology of human neurological and psychiatric diseases due to the differences between humans and rodents. The recent developments in gene manipulation of non-human primate have been attracting much attention in the biomedical research field, because non-human primates have more applicable brain structure and function than rodents. In this review, we summarize recent progress on genetically-modified non-human primates including transgenic and knockout animals using genome editing technology.

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Nestin expression is differently regulated between odontoblasts and the subodontoblastic layer in mice

Nakatomi M., Quispe-Salcedo A., Sakaguchi M., Ida-Yonemochi H., Okano H., Ohshima H.

Histochemistry and Cell Biology （Histochemistry and Cell Biology） 149 （ 4 ） 383 - 391 2018年04月

ISSN 09486143

© 2018, Springer-Verlag GmbH Germany, part of Springer Nature. The Nestin gene encodes type VI intermediate filament and is known to be expressed in undifferentiated cells during neurogenesis and myogenesis. To regulate Nestin expression, the first or second intron enhancer is activated in a tissue-dependent manner, for example, the former in mesodermal cells and the latter in neural stem cells. Although Nestin has also been used as a differentiation marker for odontoblasts during tooth development, how Nestin expression is regulated in odontoblasts remains unclear. Therefore, this study aimed to compare the expression patterns of Nestin-GFP (green fluorescent protein) with that of endogenous Nestin in developing teeth of Nestin-EGFP (enhanced GFP) transgenic mice, in which the second intron enhancer is connected with the EGFP domain, at postnatal 7d, 3w, and 8w. Immunohistochemical and in situ hybridization analyses revealed that endogenous Nestin protein and Nestin mRNA were intensely expressed in differentiated odontoblasts, while GFP immunoreactivity, which reflects the activity of Nestin second intron enhancer-mediated transcription, was mainly observed in the subodontoblastic layer. These results indicate that the first intron enhancer may be activated in differentiated odontoblasts. Intriguingly, Nestin-GFP expression in the subodontoblastic layer was found to be restricted to the coronal pulp of molars, which is susceptible to tooth injuries. Because the subodontoblastic layer serves as a reservoir of newly differentiated odontoblast-like cells upon exogenous stimuli to dentin, our findings suggest that the original odontoblasts and regenerated odontoblast-like cells may differently regulate Nestin expression.

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Generation of D1-1 TALEN isogenic control cell line from Dravet syndrome patient iPSCs using TALEN-mediated editing of the SCN1A gene

Tanaka Y., Sone T., Higurashi N., Sakuma T., Suzuki S., Ishikawa M., Yamamoto T., Mitsui J., Tsuji H., Okano H., Hirose S.

Stem Cell Research （Stem Cell Research） 28 100 - 104 2018年04月

ISSN 18735061

© 2018 Dravet syndrome (DS) is an infantile epileptic encephalopathy mainly caused by de novo mutations in the SCN1A gene encoding the α1 subunit of the voltage-gated sodium channel Nav1.1. As an in vitro model of this disease, we previously generated an induced pluripotent stem cell (iPSC) line from a patient with DS carrying a c.4933C>T (p.R1645*) substitution in SCN1A. Here, we describe developing a genome-edited control cell line from this DS iPSC line by substituting the point mutation with the wild-type residue. This artificial control iPSC line will be a powerful tool for research into the pathology of DS.

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Rostrocaudal areal patterning of human PSC-derived cortical neurons by FGF8 signaling

Imaizumi K., Fujimori K., Ishii S., Otomo A., Hosoi Y., Miyajima H., Warita H., Aoki M., Hadano S., Akamatsu W., Okano H.

eNeuro （eNeuro） 5 （ 2 ） 2018年03月

© 2018 Imaizumi et al. The cerebral cortex is subdivided into distinct areas that have particular functions. The rostrocaudal (R-C) gradient of fibroblast growth factor 8 (FGF8) signaling defines this areal identity during neural development. In this study, we recapitulated cortical R-C patterning in human pluripotent stem cell (PSC) cultures. Modulation of FGF8 signaling appropriately regulated the R-C markers, and the patterns of global gene expression resembled those of the corresponding areas of human fetal brains. Furthermore, we demonstrated the utility of this culture system in modeling the area-specific forebrain phenotypes [presumptive upper motor neuron (UMN) phenotypes] of amyotrophic lateral sclerosis (ALS). We anticipate that our culture system will contribute to studies of human neurodevelopment and neurological disease modeling.

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Digital gene atlas of neonate common marmoset brain

Shimogori T., Abe A., Go Y., Hashikawa T., Kishi N., Kikuchi S., Kita Y., Niimi K., Nishibe H., Okuno M., Saga K., Sakurai M., Sato M., Serizawa T., Suzuki S., Takahashi E., Tanaka M., Tatsumoto S., Toki M., U M., Wang Y., Windak K., Yamagishi H., Yamashita K., Yoda T., Yoshida A., Yoshida C., Yoshimoto T., Okano H.

Neuroscience Research （Neuroscience Research） 128 1 - 13 2018年03月

ISSN 01680102

© 2017 Elsevier Ireland Ltd and Japan Neuroscience Society Interest in the common marmoset (Callithrix jacchus) as a primate model animal has grown recently, in part due to the successful demonstration of transgenic marmosets. However, there is some debate as to the suitability of marmosets, compared to more widely used animal models, such as the macaque monkey and mouse. Especially, the usage of marmoset for animal models of human cognition and mental disorders, is still yet to be fully explored. To examine the prospects of the marmoset model for neuroscience research, the Marmoset Gene Atlas (https://gene-atlas.bminds.brain.riken.jp/) provides a whole brain gene expression atlas in the common marmoset. We employ in situ hybridization (ISH) to systematically analyze gene expression in neonate marmoset brains, which allows us to compare expression with other model animals such as mouse. We anticipate that these data will provide sufficient information to develop tools that enable us to reveal marmoset brain structure, function, cellular and molecular organization for primate brain research.

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Single-cell bioluminescence imaging of deep tissue in freely moving animals

Iwano S., Sugiyama M., Hama H., Watakabe A., Hasegawa N., Kuchimaru T., Tanaka K., Takahashi M., Ishida Y., Hata J., Shimozono S., Namiki K., Fukano T., Kiyama M., Okano H., Kizaka-Kondoh S., McHugh T., Yamamori T., Hioki H., Maki S., Miyawaki A.

Science （Science） 359 （ 6378 ） 935 - 939 2018年02月

ISSN 00368075

© 2017 The Authors. Bioluminescence is a natural light source based on luciferase catalysis of its substrate luciferin. We performed directed evolution on firefly luciferase using a red-shifted and highly deliverable luciferin analog to establish AkaBLI, an all-engineered bioluminescence in vivo imaging system. AkaBLI produced emissions in vivo that were brighter by a factor of 100 to 1000 than conventional systems, allowing noninvasive visualization of single cells deep inside freely moving animals. Single tumorigenic cells trapped in the mouse lung vasculature could be visualized. In the mouse brain, genetic labeling with neural activity sensors allowed tracking of small clusters of hippocampal neurons activated by novel environments. In a marmoset, we recorded video-rate bioluminescence from neurons in the striatum, a deep brain area, for more than 1 year. AkaBLI is therefore a bioengineered light source to spur unprecedented scientific, medical, and industrial applications.

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Down-regulation of ghrelin receptors on dopaminergic neurons in the substantia nigra contributes to Parkinson's disease-like motor dysfunction

Suda Y., Kuzumaki N., Sone T., Narita M., Tanaka K., Hamada Y., Iwasawa C., Shibasaki M., Maekawa A., Matsuo M., Akamatsu W., Hattori N., Okano H., Narita M.

Molecular Brain （Molecular Brain） 11 （ 1 ） 2018年02月

© 2018 The Author(s). Ghrelin exerts a wide range of physiological actions throughout the body and appears to be a promising target for disease therapy. Endogenous ghrelin receptors (GHSRs) are present in extrahypothalamic sites including the substantia nigra pars compacta (SNc), which is related to phenotypic dysregulation or frank degeneration in Parkinson's disease (PD). Here we found a dramatic decrease in the expression of GHSR in PD-specific induced pluripotent stem cell (iPSC)-derived dopaminergic (DAnergic) neurons generated from patients carrying parkin gene (PARK2) mutations compared to those from healthy controls. Consistently, a significant decrease in the expression of GHSR was found in DAnergic neurons of isogenic PARK2-iPSC lines that mimicked loss of function of the PARK2 gene through CRISPR Cas9 technology. Furthermore, either intracerebroventricular injection or microinjection into the SNc of the selective GHSR1a antagonist [D-Lys3]-GHRP6 in normal mice produced cataleptic behaviors related to dysfunction of motor coordination. These findings suggest that the down-regulation of GHSRs in SNc-DA neurons induced the initial dysfunction of DA neurons, leading to extrapyramidal disorder under PD.

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Data descriptor: The Brain/MINDS 3D digital marmoset brain atlas

Woodward A., Hashikawa T., Maeda M., Kaneko T., Hikishima K., Iriki A., Okano H., Yamaguchi Y.

Scientific Data （Scientific Data） 5 2018年02月

© Author(s) 2018. We present a new 3D digital brain atlas of the non-human primate, common marmoset monkey (Callithrix jacchus), with MRI and coregistered Nissl histology data. To the best of our knowledge this is the first comprehensive digital 3D brain atlas of the common marmoset having normalized multi-modal data, cortical and sub-cortical segmentation, and in a common file format (NIfTI). The atlas can be registered to new data, is useful for connectomics, functional studies, simulation and as a reference. The atlas was based on previously published work but we provide several critical improvements to make this release valuable for researchers. Nissl histology images were processed to remove illumination and shape artifacts and then normalized to the MRI data. Brain region segmentation is provided for both hemispheres. The data is in the NIfTI format making it easy to integrate into neuroscience pipelines, whereas the previous atlas was in an inaccessible file format. We also provide cortical, mid-cortical and white matter boundary segmentations useful for visualization and analysis.

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Elavl3 is essential for the maintenance of Purkinje neuron axons

Ogawa Y., Kakumoto K., Yoshida T., Kuwako K., Miyazaki T., Yamaguchi J., Konno A., Hata J., Uchiyama Y., Hirai H., Watanabe M., Darnell R., Okano H., Okano H.

Scientific reports （Scientific reports） 8 （ 1 ） 2018年02月

Neuronal Elav-like (nElavl or neuronal Hu) proteins are RNA-binding proteins that regulate RNA stability and alternative splicing, which are associated with axonal and synaptic structures. nElavl proteins promote the differentiation and maturation of neurons via their regulation of RNA. The functions of nElavl in mature neurons are not fully understood, although Elavl3 is highly expressed in the adult brain. Furthermore, possible associations between nElavl genes and several neurodegenerative diseases have been reported. We investigated the relationship between nElavl functions and neuronal degeneration using Elavl3-/- mice. Elavl3-/- mice exhibited slowly progressive motor deficits leading to severe cerebellar ataxia, and axons of Elavl3-/- Purkinje cells were swollen (spheroid formation), followed by the disruption of synaptic formation of axonal terminals. Deficit in axonal transport and abnormalities in neuronal polarity was observed in Elavl3-/- Purkinje cells. These results suggest that nElavl proteins are crucial for the maintenance of axonal homeostasis in mature neurons. Moreover, Elavl3-/- mice are unique animal models that constantly develop slowly progressive axonal degeneration. Therefore, studies of Elavl3-/- mice will provide new insight regarding axonal degenerative processes.

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Nitrative stress and tau accumulation in amyotrophic lateral sclerosis/parkinsonism-dementia complex (ALS/PDC) in the Kii Peninsula, Japan

Hata Y., Ma N., Yoneda M., Morimoto S., Okano H., Murayama S., Kawanishi S., Kuzuhara S., Kokubo Y.

Frontiers in Neuroscience （Frontiers in Neuroscience） 11 （ JAN ） 2018年01月

ISSN 16624548

© 2018 Hata, Ma, Yoneda, Morimoto, Okano, Murayama, Kawanishi, Kuzuhara and Kokubo. Objective: The Kii Peninsula of Japan is known to be a high incidence area of amyotrophic lateral sclerosis/parkinsonism-dementia complex (Kii ALS/PDC) with tauopathy. Nitrative stress and oxidative stress on ALS/PDC and their relationship to tau pathology were clarified. Methods: Seven patients with Kii ALS/PDC (3 males and 4 females, average age 70.7 years, 3 with ALS, 2 with ALS with dementia, and 2 with PDC) were analyzed in this study. Five patients with Alzheimer's disease and five normal aged subjects were used as controls. Immunohistochemical analysis was performed on formalin-fixed, paraffin-embedded temporal lobe sections (the hippocampal area including hippocampus, prosubiculum, subiculum, presubiculum, and parahippocampal gyri) using antibodies to detect phosphorylated tau (anti-AT-8), nitrated guanine (anti-8-NG), anti-iNOS, anti-NFκB, and oxidized guanine (anti-8-OHdG) antibodies. Results: Most hippocampal neurons of Kii ALS/PDC patients were stained with anti-8-NG, anti-iNOS, anti-NFκB, and anti-8-OHdG antibodies and some AT-8 positive neurons were co-stained with anti-8-NG antibody. The numbers of 8-NG positive neurons and 8-OHdG positive neurons were greater than AT-8 positive neurons and the number of 8-NG positive neurons was larger in patients with Kii ALS/PDC than in controls. Conclusion: Nitrative and oxidative stress may take priority over tau accumulation and lead to the neurodegeneration in Kii ALS/PDC.

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Chromatin remodeler CHD7 regulates the stem cell identity of human neural progenitors

Chai M., Sanosaka T., Okuno H., Zhou Z., Koya I., Banno S., Andoh-Noda T., Tabata Y., Shimamura R., Hayashi T., Ebisawa M., Sasagawa Y., Nikaido I., Okano H., Kohyama J.

Genes and Development （Genes and Development） 32 （ 2 ） 165 - 180 2018年01月

ISSN 08909369

© 2018 Chai et al. Multiple congenital disorders often present complex phenotypes, but how the mutation of individual genetic factors can lead to multiple defects remains poorly understood. In the present study, we used human neuroepithelial (NE) cells and CHARGE patient-derived cells as an in vitro model system to identify the function of chromodomain helicase DNA-binding 7 (CHD7) in NE–neural crest bifurcation, thus revealing an etiological link between the central nervous system (CNS) and craniofacial anomalies observed in CHARGE syndrome. We found that CHD7 is required for epigenetic activation of superenhancers and CNS-specific enhancers, which support the maintenance of the NE and CNS lineage identities. Furthermore, we found that BRN2 and SOX21 are downstream effectors of CHD7, which shapes cellular identities by enhancing a CNS-specific cellular program and indirectly repressing non-CNS-specific cellular programs. Based on our results, CHD7, through its interactions with superenhancer elements, acts as a regulatory hub in the orchestration of the spatiotemporal dynamics of transcription factors to regulate NE and CNS lineage identities.

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Activation of ventral tegmental area dopaminergic neurons reverses pathological allodynia resulting from nerve injury or bone cancer

Watanabe M., Narita M., Hamada Y., Yamashita A., Tamura H., Ikegami D., Kondo T., Shinzato T., Shimizu T., Fukuchi Y., Muto A., Okano H., Yamanaka A., Tawfik V., Kuzumaki N., Navratilova E., Porreca F., Narita M.

Molecular Pain （Molecular Pain） 14 2018年01月

© The Author(s) 2018. Chronic pain induced by nerve damage due to trauma or invasion of cancer to the bone elicits severe ongoing pain as well as hyperalgesia and allodynia likely reflecting adaptive changes within central circuits that amplify nociceptive signals. The present study explored the possible contribution of the mesolimbic dopaminergic circuit in promoting allodynia related to neuropathic and cancer pain. Mice with ligation of the sciatic nerve or treated with intrafemoral osteosarcoma cells showed allodynia to a thermal stimulus applied to the paw on the injured side. Patch clamp electrophysiology revealed that the intrinsic neuronal excitability of ventral tegmental area (VTA) dopamine neurons projecting to the nucleus accumbens (N.Acc.) was significantly reduced in those mice. We used tyrosine hydroxylase (TH)-cre mice that were microinjected with adeno-associated virus (AAV) to express channelrhodopsin-2 (ChR2) to allow optogenetic stimulation of VTA dopaminergic neurons in the VTA or in their N.Acc. terminals. Optogenetic activation of these cells produced a significant but transient anti-allodynic effect in nerve injured or tumor-bearing mice without increasing response thresholds to thermal stimulation in sham-operated animals. Suppressed activity of mesolimbic dopaminergic neurons is likely to contribute to decreased inhibition of N.Acc. output neurons and to neuropathic or cancer pain-induced allodynia suggesting strategies for modulation of pathological pain states.

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Expression pattern of EYA4 in the common marmoset (Callithrix jacchus) cochlea

Matsuzaki S., Hosoya M., Okano H., Fujioka M., Ogawa K.

Neuroscience Letters （Neuroscience Letters） 662 185 - 188 2018年01月

ISSN 03043940

© 2017 The eyes absent (EYA)-like genes are essential for the formation of sensory organs among fly (Drosophila melanogaster) and mammals. EYA4, one of the vertebrate genes of Eya family, is reported to be causative for late-onset mid-frequency sensorineural hearing loss in humans, while Eya4-deficient mice exhibited congenital profound deafness and otitis media with effusion due to the eustachian tube dysmorphology. Because of the species difference in the phenotype, the pathophysiology of EYA4 in the human cochlea has yet to be elucidated. Here, we examine the expression pattern of EYA4 in the cochlea of common marmoset (Callithrix jacchus), a non-human primate. The results indicated a distinct expression pattern of EYA4 in the adult marmoset cochleae, especially strong in all supporting cells, while in mouse their expressions were diminished. Interestingly, EYA4 expression in the hair cells, supporting cells and neurons was co-localized with sine oculis homeobox–SIX1, a transcription factor essential for the transcriptional activity of EYA4. The results revealed inter-species differences in the expression pattern of EYA4 gene in supporting cells between primates and rodents. The results also indicated a fundamental role of EYA4 in the primate auditory cells. Experiments with primate models such as marmosets or with human cochlear cells may provide cues about the unknown pathogenesis of EYA4-related hearing loss.

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Parkin absence accelerates microtubule aging in dopaminergic neurons

Cartelli D., Amadeo A., Calogero A., Casagrande F., De Gregorio C., Gioria M., Kuzumaki N., Costa I., Sassone J., Ciammola A., Hattori N., Okano H., Goldwurm S., Roybon L., Pezzoli G., Cappelletti G.

Neurobiology of Aging （Neurobiology of Aging） 61 66 - 74 2018年01月

ISSN 01974580

© 2017 Elsevier Inc. Loss-of-function caused by mutations in the parkin gene (PARK2) lead to early-onset familial Parkinson's disease. Recently, mechanistic studies proved the ability of parkin in regulating mitochondria homeostasis and microtubule (MT) stability. Looking at these systems during aging of PARK2 knockout mice, we found that loss of parkin induced an accelerated (over)acetylation of MT system both in dopaminergic neuron cell bodies and fibers, localized in the substantia nigra and corpus striatum, respectively. Interestingly, in PARK2 knockout mice, changes of MT stability preceded the alteration of mitochondria transport. Moreover, in-cell experiments confirmed that loss of parkin affects mitochondria mobility and showed that this defect depends on MT system as it is rescued by paclitaxel, a well-known MT-targeted agent. Furthermore, both in PC12 neuronal cells and in patients' induced pluripotent stem cell–derived midbrain neurons, we observed that parkin deficiencies cause the fragmentation of stable MTs. Therefore, we suggest that parkin acts as a regulator of MT system during neuronal aging, and we endorse the hypothesis that MT dysfunction may be crucial in the pathogenesis of Parkinson's disease.

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Chemical decontamination of iPS cell-derived neural cell mixtures

Mao D., Chung X., Andoh-Noda T., Qin Y., Sato S., Takemoto Y., Akamatsu W., Okano H., Uesugi M.

Chemical Communications （Chemical Communications） 54 （ 11 ） 1355 - 1358 2018年

ISSN 13597345

© 2018 The Royal Society of Chemistry. This report describes the design and evaluation of phosphorylated 7-ethyl-10-hydroxycamptothecin (SN38-P), which selectively eliminates tumor-forming proliferative stem cells, including human induced pluripotent stem cells (hiPSCs) and neural stem cells, from iPSC-derived neural cell mixtures. Results of the present study demonstrate that simple phosphorylation of an anticancer drug can provide a safe, cost-effective, and chemically-defined tool for decontaminating hiPSC-derived neuron.

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Astrocyte-derived exosomes treated with a semaphorin 3A inhibitor enhance stroke recovery via prostaglandin D <inf>2</inf> synthase

Hira K., Ueno Y., Tanaka R., Miyamoto N., Yamashiro K., Inaba T., Urabe T., Okano H., Hattori N.

Stroke （Stroke） 49 （ 10 ） 2483 - 2494 2018年

ISSN 00392499

© 2018 American Heart Association, Inc. Background and Purpose: Exosomes play a pivotal role in neurogenesis. In the peri-infarct area after stroke, axons begin to regenerate but are inhibited by astrocyte scar formation. The direct effect and underlying molecular mechanisms of astrocyte-derived exosomes on axonal outgrowth after ischemia are not known. Methods: Using a semaphorin 3A (Sema3A) inhibitor, we explored neuronal signaling during axonal outgrowth after ischemia in rats subjected to middle cerebral artery occlusion and in cultured cortical neurons challenged with oxygen-glucose deprivation. Furthermore, we assessed whether this inhibitor suppressed astrocyte activation and regulated astrocyte-derived exosomes to enhance axonal outgrowth after ischemia. Results: In rats subjected to middle cerebral artery occlusion, we administered a Sema3A inhibitor into the peri-infarct area from 7 to 21 days after occlusion. We found that phosphorylated high-molecular weight neurofilament-immunoreactive axons were increased, glial fibrillary acidic protein-immunoreactive astrocytes were decreased, and functional recovery was promoted at 28 days after middle cerebral artery occlusion. In cultured neurons, the Sema3A inhibitor decreased Rho family GTPase 1, increased R-Ras, which phosphorylates Akt and glycogen synthase kinase 3β (GSK-3β), selectively increased phosphorylated GSK-3β in axons, and thereby enhanced phosphorylated high-molecular weight neurofilament-immunoreactive axons after oxygen-glucose deprivation. In cultured astrocytes, the Sema3A inhibitor suppressed activation of astrocytes induced by oxygen-glucose deprivation. Exosomes secreted from ischemic astrocytes treated with the Sema3A inhibitor further promoted axonal elongation and increased prostaglandin D 2 synthase expression on microarray analysis. GSK-3β + and prostaglandin D 2 synthase + neurons were robustly increased after treatment with the Sema3A inhibitor in the peri-infarct area. Conclusions: Neuronal Rho family GTPase 1/R-Ras/Akt/GSK-3β signaling, axonal GSK-3β expression, and astrocyte-derived exosomes with prostaglandin D 2 synthase expression contribute to axonal outgrowth and functional recovery after stroke.

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Reactive astrocytes function as phagocytes after brain ischemia via ABCA1-mediated pathway

Morizawa Yosuke M., Hirayama Yuri, Ohno Noubuhiko, Shibata Shinsuke, Shigetomi Eiji, Sui Yang, Nabekura Junichi, Sato Koichi, Okajima Fumikazu, Takebayashi Hirohide, Okano Hideyuki, Koizumi Schuichi

Nature Communications 8 （ 1 ） 2017年12月

ISSN 2041-1723

Astrocytes become reactive following various brain insults; however, the functions of reactive astrocytes are poorly understood. Here, we show that reactive astrocytes function as phagocytes after transient ischemic injury and appear in a limited spatiotemporal pattern. Following transient brain ischemia, phagocytic astrocytes are observed within the ischemic penumbra region during the later stage of ischemia. However, phagocytic microglia are mainly observed within the ischemic core region during the earlier stage of ischemia. Phagocytic astrocytes upregulate ABCA1 and its pathway molecules, MEGF10 and GULP1, which are required for phagocytosis, and upregulation of ABCA1 alone is sufficient for enhancement of phagocytosis in vitro. Disrupting ABCA1 in reactive astrocytes result in fewer phagocytic inclusions after ischemia. Together, these findings suggest that astrocytes are transformed into a phagocytic phenotype as a result of increase in ABCA1 and its pathway molecules and contribute to remodeling of damaged tissues and penumbra networks.

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Regulation of Sema3c and the Interaction between Cardiac Neural Crest and Second Heart Field during Outflow Tract Development

Kodo Kazuki, Shibata Shinsuke, Miyagawa-Tomita Sachiko, Ong Sang Ging, Takahashi Hiroshi, Kume Tsutomu, Okano Hideyuki, Matsuoka Rumiko, Yamagishi Hiroyuki

Scientific Reports 7 （ 1 ） 2017年12月

ISSN 2045-2322

The cardiac neural crest cells (cNCCs) and the second heart field (SHF) play key roles in development of the cardiac outflow tract (OFT) for establishment of completely separated pulmonary and systemic circulations in vertebrates. A neurovascular guiding factor, Semaphorin 3c (Sema3c), is required for the development of the OFT, however, its regulation of the interaction between cNCCs and SHF remains to be determined. Here, we show that a Sema3c is a candidate that mediates interaction between cNCCs and the SHF during development of the OFT. Foxc1/c2 directly activates the transcription of Sema3c in the OFT, whereas, a hypomorph of Tbx1, a key SHF transcription factor, resulted in the ectopic expression of Sema3c in the pharyngeal arch region. Fgf8, a downstream secreted factor of Tbx1, inhibited the expression of Sema3c in cNCCs via activation of ERK1/2 signaling. Blocking of FGF8 caused ectopic expression of SEMA3C and a migration defect of cNCCs, resulting in abnormal chick pharyngeal arch development. These results suggest that proper spatio-temporal expression of Sema3c, regulated positively by Foxc1/c2 and negatively by the Tbx1-Fgf8 cascade, respectively, is essential for the interaction between cNCCs and the SHF that correctly navigates cNCCs towards the OFT, composed of SHF-derived cells.

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Visualization of migration of human cortical neurons generated from induced pluripotent stem cells

Bamba Yohei, Kanemura Yonehiro, Okano Hideyuki, Yamasaki Mami

Journal of Neuroscience Methods 289 57 - 63 2017年09月

ISSN 0165-0270

Background Neuronal migration is considered a key process in human brain development. However, direct observation of migrating human cortical neurons in the fetal brain is accompanied by ethical concerns and is a major obstacle in investigating human cortical neuronal migration. New method We established a novel system that enables direct visualization of migrating cortical neurons generated from human induced pluripotent stem cells (hiPSCs). Results We observed the migration of cortical neurons generated from hiPSCs derived from a control and from a patient with lissencephaly. Methods Our system needs no viable brain tissue, which is usually used in slice culture. Migratory behavior of human cortical neuron can be observed more easily and more vividly by its fluorescence and glial scaffold than that by earlier methods. Conclusions Our in vitro experimental system provides a new platform for investigating development of the human central nervous system and brain malformation.

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Evidence that phosphorylated ubiquitin signaling is involved in the etiology of Parkinson's disease

Shiba-Fukushima Kahori, Ishikawa Kei Ichi, Inoshita Tsuyoshi, Izawa Nana, Takanashi Masashi, Sato Shigeto, Onodera Osamu, Akamatsu Wado, Okano Hideyuki, Imai Yuzuru, Hattori Nobutaka

Human Molecular Genetics 26 （ 16 ） 3172 - 3185 2017年08月

ISSN 0964-6906

The ubiquitin (Ub) kinase PINK1 and the E3 Ub ligase Parkin, two gene products associated with young-onset Parkinson's disease (PD), participate in mitochondrial quality control. The phosphorylation of mitochondrial polyUb by PINK1, which is activated in a mitochondrial membrane potential (ΔΨm)-dependent manner, facilitates the mitochondrial translocation and concomitant enzymatic activation of Parkin, leading to the clearance of phospho-polyUb-tagged mitochondria via mitophagy. Thus, Ub phosphorylation is a key event in PINK1-Parkin-mediated mitophagy. Here, we examined the role of phospho-Ub signaling in the pathogenesis of PD using fly PD models, human brain tissue and dopaminergic neurons derived from induced pluripotent stem cells (iPSCs) containing Parkin or PINK1 mutations, as well as normal controls.We report that phospho-Ub signaling is highly conserved between humans and Drosophila, and that phospho-Ub signaling and the relocation of axonal mitochondria upon DWmreduction are indeed compromised in human dopaminergic neurons containing Parkin or PINK1 mutations. Moreover, phospho-Ub signaling is prominent in tyrosine hydroxylase-positive neurons compared with tyrosine hydroxylase-negative neurons, suggesting that PINK1-Parkin signaling is more required for dopaminergic neurons. These results shed light on the particular vulnerability of dopaminergic neurons to mitochondrial stress.

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MIF: functions in brain and glioblastoma

Ohta Shigeki, Kawakami Yutaka, Okano Hideyuki

Oncotarget 8 （ 29 ） 46706 - 46707 2017年06月

ISSN 1949-2553

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Hypoxia Epigenetically Confers Astrocytic Differentiation Potential on Human Pluripotent Cell-Derived Neural Precursor Cells

Yasui Tetsuro, Uezono Naohiro, Nakashima Hideyuki, Noguchi Hirofumi, Matsuda Taito, Noda-Andoh Tomoko, Okano Hideyuki, Nakashima Kinichi

Stem Cell Reports 8 （ 6 ） 1743 - 1756 2017年06月

ISSN 2213-6711

Human neural precursor cells (hNPCs) derived from pluripotent stem cells display a high propensity for neuronal differentiation, but they require long-term culturing to differentiate efficiently into astrocytes. The mechanisms underlying this biased fate specification of hNPCs remain elusive. Here, we show that hypoxia confers astrocytic differentiation potential on hNPCs through epigenetic gene regulation, and that this was achieved by cooperation between hypoxia-inducible factor 1α and Notch signaling, accompanied by a reduction of DNA methylation level in the promoter region of a typical astrocyte-specific gene, Glial fibrillary acidic protein. Furthermore, we found that this hypoxic culture condition could be applied to rapid generation of astrocytes from Rett syndrome patient-derived hNPCs, and that these astrocytes impaired neuronal development. Thus, our findings shed further light on the molecular mechanisms regulating hNPC differentiation and provide attractive tools for the development of therapeutic strategies for treating astrocyte-mediated neurological disorders.

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A noncoding RNA containing a SINE-B1 motif associates with meiotic metaphase chromatin and has an indispensable function during spermatogenesis

Nakajima Ryusuke, Sato Takuya, Ogawa Takehiko, Okano Hideyuki, Noce Toshiaki

PLoS ONE 12 （ 6 ） 2017年06月

ISSN 1932-6203

A search for early response genes that are activated following germ cell induction from mouse embryonic stem cells in vitro led us to the isolation of a long noncoding RNA that contains a SINE (short interspersed element)-B1F motif that was named R53. In situ hybridization and northern blot analyses revealed that the R53 subfragment RNA bears a B1F motif, is processed from the primary transcript, is expressed in adult testis and is predominantly localized in meiotic metaphase chromatin during spermatogenesis. Recent studies of chromosome- associated RNAs have explored novel functions of noncoding RNAs. Specifically, chromosome-bound noncoding RNAs function not only as structural components of chromosome but also as scaffolds that recruit epigenetic modulators for transcriptional regulation, and they are dynamically rearranged during the cell cycle. However, few studies have explored meiotic chromatin; thus, R53 RNA appears to be the first long noncoding RNA to be tightly associated with the metaphase chromatin during spermatogenesis. Furthermore, R53 knockdown using a lentivirus-mediated RNAi injected into mouse testis and organ culture of the fragments revealed a remarkable reduction in postmeiotic cells and irregular upregulation of several postmeiotic genes, which suggests the possibility that the SINE-B1- derived noncoding RNA R53 plays an indispensable role in the transcriptional regulation of key spermatogenesis genes.

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Developmental trajectory of the corpus callosum from infancy to the juvenile stage

Sakai Tomoko, Mikami Akichika, Suzuki Juri, Miyabe-Nishiwaki Takako, Matsui Mie, Tomonaga Masaki, Hamada Yuzuru, Matsuzawa Tetsuro, Okano Hideyuki, Oishi Kenichi

PLoS ONE 12 （ 6 ） 2017年06月

ISSN 1932-6203

How brains develop during early life is one of the most important topics in neuroscience because it underpins the neuronal functions that mature during this period. A comparison of the neurodevelopmental patterns among humans and nonhuman primates is essential to infer evolutional changes in neuroanatomy that account for higher-order brain functions, especially those specific to humans. The corpus callosum (CC) is the major white matter bundle that connects the cerebral hemispheres, and therefore, relates to a wide variety of neuronal functions. In humans, the CC area rapidly expands during infancy, followed by relatively slow changes. In chimpanzees, based on a cross-sectional study, slow changes in the CC area during the juvenile stage and later have also been reported. However, little is known about the developmental changes during infancy. A longitudinal study is also required to validate the previous cross-sectional observations about the chimpanzee CC. The present longitudinal study of magnetic resonance imaging scans demonstrates that the CC development in chimpanzees and humans is characterized by a rapid increase during infancy, followed by gradual increase during the juvenile stage. Several differences between the two species were also identified. First, there was a tendency toward a greater increase in the CC areas during infancy in humans. Second, there was a tendency toward a greater increase in the rostrum during the juvenile stage in chimpanzees. The rostral body is known to carry fibers between the bilateral prefrontal and premotor cortices, and is involved in behavior planning and control, verbal working memory, and number conception. The rostrum is known to carry fibers between the prefrontal cortices, and is involved in attention control. The interspecies differences in the developmental trajectories of the rostral body and the rostrum might be related to evolutional changes in the brain systems.

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Muscle architectural properties in the common marmoset (Callithrix jacchus)

Ogihara Naomichi, Oishi Motoharu, Kanai Ryogo, Shimada Hikaru, Kondo Takahiro, Yoshino-Saito Kimika, Ushiba Junichi, Okano Hideyuki

Primates 1 - 12 2017年05月

ISSN 0032-8332

The common marmoset, Callithrix jacchus, is a small New World monkey that has recently gained attention as an important experimental animal model in the field of neuroscience as well in rehabilitative and regenerative medicine. This attention reflects the closer phylogenetic relationship between humans and common marmosets compared to that between humans and other experimental animals. When studying the neuronal mechanism behind various types of neurological motor disorders using the common marmoset, possible differences in muscle parameters (e.g., the force-generating capacity of each of the muscles) between the common marmoset and other animals must be taken into account to permit accurate interpretation of observed motor behavior. Differences in the muscle architectural properties are expected to affect biomechanics, and hence to affect neuronal control of body movements. Therefore, we dissected the forelimbs and hind limbs of two common marmosets, including systematic analysis of the muscle mass, fascicle length, and physiological cross-sectional area (PCSA). Comparisons of the mass fractions and PCSA fractions of the forelimb and hind limb musculature among the common marmoset, human, Japanese macaque, and domestic cat demonstrated that the overall muscle architectural properties of the forelimbs and hind limbs in the common marmoset are very similar to those of the Japanese macaque, a typical quadrupedal primate. However, muscle architectural properties of the common marmoset differ from those of the domestic cat, which has relatively larger hamstrings and pedal digital flexor muscles. Compared to humans, the common marmoset exhibits relatively smaller shoulder protractor, retractor, and abductor muscles and larger elbow extensor and rotator-cuff muscles in the forelimb, and smaller plantarflexor muscles in the hind limb. These differences in the muscle architectural properties must be taken into account when interpreting motor behaviors such as locomotion and arm-reaching movements in the common marmoset.

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Fail-Safe System against Potential Tumorigenicity after Transplantation of iPSC Derivatives

Itakura Go, Kawabata Soya, Ando Miki, Nishiyama Yuichiro, Sugai Keiko, Ozaki Masahiro, Iida Tsuyoshi, Ookubo Toshiki, Kojima Kota, Kashiwagi Rei, Yasutake Kaori, Nakauchi Hiromitsu, Miyoshi Hiroyuki, Nagoshi Narihito, Kohyama Jun, Iwanami Akio, Matsumoto Morio, Nakamura Masaya, Okano Hideyuki

Stem Cell Reports 8 （ 3 ） 673 - 684 2017年03月

ISSN 2213-6711

Human induced pluripotent stem cells (iPSCs) are promising in regenerative medicine. However, the risks of teratoma formation and the overgrowth of the transplanted cells continue to be major hurdles that must be overcome. Here, we examined the efficacy of the inducible caspase-9 (iCaspase9) gene as a fail-safe against undesired tumorigenic transformation of iPSC-derived somatic cells. We used a lentiviral vector to transduce iCaspase9 into two iPSC lines and assessed its efficacy in vitro and in vivo. In vitro, the iCaspase9 system induced apoptosis in approximately 95% of both iPSCs and iPSC-derived neural stem/progenitor cells (iPSC-NS/PCs). To determine in vivo function, we transplanted iPSC-NS/PCs into the injured spinal cord of NOD/SCID mice. All transplanted cells whose mass effect was hindering motor function recovery were ablated upon transduction of iCaspase9. Our results suggest that the iCaspase9 system may serve as an important countermeasure against post-transplantation adverse events in stem cell transplant therapies.

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Skin-derived precursors as a source of progenitors for corneal endothelial regeneration

Inagaki Emi, Hatou Shin, Higa Kazunari, Yoshida Satoru, Shibata Shinsuke, Okano Hideyuki, Tsubota Kazuo, Shimmura Shigeto

Stem Cells Translational Medicine 6 （ 3 ） 788 - 798 2017年03月

ISSN 2157-6564

Corneal blindness is the fourth leading cause of blindness in the world. Current treatment is allogenic corneal transplantation, which is limited by shortage of donors and immunological rejection. Skin-derived precursors (SKPs) are postnatal stem cells, which are self-renewing, multipotent precursors that can be isolated and expanded from the dermis. Facial skin may therefore be an accessible autologous source of neural crest derived cells. SKPs were isolated from facial skin of Wnt1- Cre/Floxed EGFP mouse. After inducing differentiation with medium containing retinoic acid and GSK 3-β inhibitor, SKPs formed polygonal corneal endothelial-like cells (sTECE). Expression of major corneal endothelial markers were confirmed by Reverse transcription polymerase chain reaction (RT-PCR) and quantitative Real time polymerase chain reaction (qRT-PCR).Western blots confirmed the expression of Na, K-ATPase protein, the major functional marker of corneal endothelial cells. Immunohistochemistry revealed the expression of zonular occludens-1 and Na, K-ATPase in cellcell junctions. In vitro functional analysis of Na, K-ATPase pump activity revealed that sTECE had significantly high pump function compared to SKPs or control 3T3 cells. Moreover, sTECE transplanted into a rabbit model of bullous keratopathy successfully maintained corneal thickness and transparency. Furthermore, we successfully induced corneal endothelial-like cells from human SKPs, and showed that transplanted corneas also maintained corneal transparency and thickness. Our findings suggest that SKPs may be used as a source of autologous cells for the treatment of corneal endothelial disease.

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Evaluation of the immunogenicity of human iPS cell-derived neural stem/progenitor cells in vitro

Ozaki Masahiro, Iwanami Akio, Nagoshi Narihito, Kohyama Jun, Itakura Go, Iwai Hiroki, Nishimura Soraya, Nishiyama Yuichiro, Kawabata Soya, Sugai Keiko, Iida Tsuyoshi, Matsubayashi Kohei, Isoda Miho, Kashiwagi Rei, Toyama Yoshiaki, Matsumoto Morio, Okano Hideyuki, Nakamura Masaya

Stem Cell Research 19 128 - 138 2017年03月

ISSN 1873-5061

To achieve the goal of a first-in-human trial for human induced pluripotent stem cell (hiPSC)-based transplantation for the treatment of various diseases, allogeneic human leukocyte antigen (HLA)-matched hiPSC cell banks represent a realistic tool from the perspective of quality control and cost performance. Furthermore, considering the limited therapeutic time-window for acute injuries, including neurotraumatic injuries, an iPS cell bank is of potential interest. However, due to the relatively immunoprivileged environment of the central nervous system, it is unclear whether HLA matching is required in hiPSC-derived neural stem/progenitor cell (hiPSC-NS/PC) transplantation for the treatment of neurodegenerative diseases and neurotraumatic injuries. In this study, we evaluated the significance of HLA matching in hiPSC-NS/PC transplantation by performing modified mixed lymphocyte reaction (MLR) assays with hiPSC-NS/PCs. Compared to fetus-derived NS/PCs, the expression levels of human leukocyte antigen-antigen D related (HLA-DR) and co-stimulatory molecules on hiPSC-NS/PCs were significantly low, even with the addition of tumor necrosis factor-α (TNFα) and/or interferon-γ (IFNγ) to mimic the inflammatory environment surrounding transplanted hiPSC-NS/PCs in injured tissues. Interestingly, both the allogeneic HLA-matched and the HLA-mismatched responses were similarly low in the modified MLR assay. Furthermore, the autologous response was also similar to the allogeneic response. hiPSC-NS/PCs suppressed the proliferative responses of allogeneic HLA-mismatched peripheral blood mononuclear cells (PBMCs) in a dose-dependent manner. Thus, the low antigen-presenting function and immunosuppressive effects of hiPSC-NS/PCs result in a depressed immune response, even in an allogeneic HLA-mismatched setting. It is crucial to verify whether these in vitro results are reproducible in a clinical setting.

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Differential gene expression profiles in neurons generated from lymphoblastoid B-cell line-derived iPS cells from monozygotic twin cases with treatment-resistant schizophrenia and discordant responses to clozapine

Nakazawa Takanobu, Kikuchi Masataka, Ishikawa Mitsuru, Yamamori Hidenaga, Nagayasu Kazuki, Matsumoto Takuya, Fujimoto Michiko, Yasuda Yuka, Fujiwara Mikiya, Okada Shota, Matsumura Kensuke, Kasai Atsushi, Hayata-Takano Atsuko, Shintani Norihito, Numata Shusuke, Takuma Kazuhiro, Akamatsu Wado, Okano Hideyuki, Nakaya Akihiro, Hashimoto Hitoshi, Hashimoto Ryota

Schizophrenia Research 181 75 - 82 2017年03月

ISSN 0920-9964

Schizophrenia is a chronic psychiatric disorder with complex genetic and environmental origins. While many antipsychotics have been demonstrated as effective in the treatment of schizophrenia, a substantial number of schizophrenia patients are partially or fully unresponsive to the treatment. Clozapine is the most effective antipsychotic drug for treatment-resistant schizophrenia; however, clozapine has rare but serious side-effects. Furthermore, there is inter-individual variability in the drug response to clozapine treatment. Therefore, the identification of the molecular mechanisms underlying the action of clozapine and drug response predictors is imperative. In the present study, we focused on a pair of monozygotic twin cases with treatment-resistant schizophrenia, in which one twin responded well to clozapine treatment and the other twin did not. Using induced pluripotent stem (iPS) cell-based technology, we generated neurons from iPS cells derived from these patients and subsequently performed RNA-sequencing to compare the transcriptome profiles of the mock or clozapine-treated neurons. Although, these iPS cells similarly differentiated into neurons, several genes encoding homophilic cell adhesion molecules, such as protocadherin genes, showed differential expression patterns between these two patients. These results, which contribute to the current understanding of the molecular mechanisms of clozapine action, establish a new strategy for the use of monozygotic twin studies in schizophrenia research.

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Correlation of insulin resistance and motor function in spinal and bulbar muscular atrophy

Nakatsuji Hideaki, Araki Amane, Hashizume Atsushi, Hijikata Yasuhiro, Yamada Shinichiro, Inagaki Tomonori, Suzuki Keisuke, Banno Haruhiko, Suga Noriaki, Okada Yohei, Ohyama Manabu, Nakagawa Tohru, Kishida Ken, Funahashi Tohru, Shimomura Iichiro, Okano Hideyuki, Katsuno Masahisa, Sobue Gen

Journal of Neurology 1 - 9 2017年02月

ISSN 0340-5354

This study aimed to evaluate various metabolic parameters in patients with spinal and bulbar muscular atrophy (SBMA), to investigate the association between those indices and disease severity, and to explore the underlying molecular pathogenesis. We compared the degree of obesity, metabolic parameters, and blood pressure in 55 genetically confirmed SBMA patients against those in 483 age- and sex-matched healthy control. In SBMA patients, we investigated the correlation between these factors and motor functional indices. SBMA patients had lower body mass index, blood glucose, and Hemoglobin A1c, but higher blood pressure, homeostasis model assessment of insulin resistance (HOMA-IR, a marker of insulin resistance), total cholesterol, and adiponectin levels than the control subjects. There were no differences in visceral fat areas, high-density lipoprotein-cholesterol (HDL-C), or triglyceride levels in two groups. Revised amyotrophic lateral sclerosis functional rating scale (ALSFRS-R) correlated positively with HDL-C, but negatively with HOMA-IR. Through stepwise multiple regression analysis, we identified HOMA-IR as a significant metabolic determinant of ALSFRS-R. In biochemical analysis, we found that decreased expressions of insulin receptors, insulin receptor substrate-1 and insulin receptor-β, in autopsied muscles and fibroblasts of SBMA patients. This study demonstrates that SBMA patients have insulin resistance, which is associated with the disease severity. The expressions of insulin receptors are attenuated in the skeletal muscle of SBMA, providing a possible pathomechanism of metabolic alterations. These findings suggested that insulin resistance is a metabolic index reflecting disease severity and pathogenesis as well as a potential therapeutic target for SBMA.

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Induction of hair follicle dermal papilla cell properties in human induced pluripotent stem cell-derived multipotent LNGFR(+)THY-1(+) mesenchymal cells

Veraitch Ophelia, Mabuchi Yo, Matsuzaki Yumi, Sasaki Takashi, Okuno Hironobu, Tsukashima Aki, Amagai Masayuki, Okano Hideyuki, Ohyama Manabu

Scientific Reports 7 2017年02月

ISSN 2045-2322

The dermal papilla (DP) is a specialised mesenchymal component of the hair follicle (HF) that plays key roles in HF morphogenesis and regeneration. Current technical difficulties in preparing trichogenic human DP cells could be overcome by the use of highly proliferative and plastic human induced pluripotent stem cells (hiPSCs). In this study, hiPSCs were differentiated into induced mesenchymal cells (iMCs) with a bone marrow stromal cell phenotype. A highly proliferative and plastic LNGFR(+)THY-1(+) subset of iMCs was subsequently programmed using retinoic acid and DP cell activating culture medium to acquire DP properties. The resultant cells (induced DP-substituting cells [iDPSCs]) exhibited up-regulated DP markers, interacted with human keratinocytes to up-regulate HF related genes, and when co-grafted with human keratinocytes in vivo gave rise to fibre structures with a hair cuticle-like coat resembling the hair shaft, as confirmed by scanning electron microscope analysis. Furthermore, iDPSCs responded to the clinically used hair growth reagent, minoxidil sulfate, to up-regulate DP genes, further supporting that they were capable of, at least in part, reproducing DP properties. Thus, LNGFR(+)THY-1(+) iMCs may provide material for HF bioengineering and drug screening for hair diseases.

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q-Space Myelin Map imaging for longitudinal analysis of demyelination and remyelination in multiple sclerosis patients treated with fingolimod

Tanikawa Mariko, Nakahara Jin, Hata Junichi, Suzuki Shigeaki, Fujiyoshi Kanehiro, Fujiwara Hirokazu, Momoshima Suketaka, Jinzaki Masahiro, Nakamura Masaya, Okano Hideyuki, Takahashi Shinichi, Suzuki Norihiro

Journal of the Neurological Sciences 373 352 - 357 2017年02月

ISSN 0022-510X

Background Fingolimod (FTY) is an oral sphingosine-1-phosphate receptor modulator that reduces relapse and slows brain atrophy in multiple sclerosis (MS) patients. In addition, FTY has been shown to enhance remyelination in certain animal models. Objective To analyze feasibility of a novel q-space Myelin Map imaging to monitor demyelination and remyelination under FTY treatment in MS patients. Methods Treatment outcomes of 24 consecutive MS patients treated with FTY were analyzed. A longitudinal analysis of q-space Myelin Map imaging was performed in a subset of these patients. Results During the treatment course (average of 16.1 months), 10 patients (42%) exhibited improvement on the Expanded Disability Status Scale (EDSS) or maintained disability-free state (“optimal responders”). The average baseline age and EDSS score were significantly younger and milder in optimal responders compared to the rest of patients. A pilot longitudinal q-space Myelin Map study in 8 patients (including 4 optimal responders) showed that optimal responders tended to show signs of remyelination while exhibiting no newly evolved demyelinated lesions. Conclusion FTY may improve disability in younger patients with milder MS, and absence of demyelination activity and presence of remyelination activity may in part be associated with such improvement. q-Space Myelin Map imaging is a clinically feasible modality to monitor demyelination and remyelination in MS patients.

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Elucidation of developmental patterns of marmoset corpus callosum through a comparative MRI in marmosets, chimpanzees, and humans

Sakai Tomoko, Komaki Yuji, Hata Junichi, Okahara Junko, Okahara Norio, Inoue Takashi, Mikami Akichika, Matsui Mie, Oishi Kenichi, Sasaki Erika, Okano Hideyuki

Neuroscience Research 2017年02月

ISSN 0168-0102

The corpus callosum (CC) is present in all primate brains and is the major white matter tract connecting the cerebral hemispheres for integration of sensory, motor and higher-order cognitive information. The midsagittal area of the CC has frequently been used as a sensitive biomarker of brain development. Although the marmoset has been considered as an alternative non-human primate model for neuroscience research, the developmental patterns of the CC have not been explored. The present longitudinal study of magnetic resonance imaging demonstrated that marmosets show a rapid increase of CC during infancy, followed by a slow increase during the juvenile stage, as observed in chimpanzees and humans. Marmosets also show a tendency toward a greater increase in CC during late infancy and the juvenile stage, as observed in humans, but not in chimpanzees. However, several differences between marmosets and humans were identified. There was a tendency toward a greater maturation of the human CC during early infancy. Furthermore, there was a tendency toward a greater increase during late infancy and the juvenile stage in marmosets, compared to that observed in chimpanzees and humans. These differences in the developmental trajectories of the CC may be related to evolutional changes in social behavior.

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Dysfunction of ventrolateral striatal dopamine receptor type 2-expressing medium spiny neurons impairs instrumental motivation

Tsutsui-Kimura Iku, Takiue Hiroyuki, Yoshida Keitaro, Xu Ming, Yano Ryutaro, Ohta Hiroyuki, Nishida Hiroshi, Bouchekioua Youcef, Okano Hideyuki, Uchigashima Motokazu, Watanabe Masahiko, Takata Norio, Drew Michael R., Sano Hiromi, Mimura Masaru, Tanaka Kenji F.

Nature Communications 8 2017年02月

ISSN 2041-1723

Impaired motivation is present in a variety of neurological disorders, suggesting that decreased motivation is caused by broad dysfunction of the nervous system across a variety of circuits. Based on evidence that impaired motivation is a major symptom in the early stages of Huntington's disease, when dopamine receptor type 2-expressing striatal medium spiny neurons (D2-MSNs) are particularly affected, we hypothesize that degeneration of these neurons would be a key node regulating motivational status. Using a progressive, time-controllable, diphtheria toxin-mediated cell ablation/dysfunction technique, we find that loss-of-function of D2-MSNs within ventrolateral striatum (VLS) is sufficient to reduce goal-directed behaviours without impairing reward preference or spontaneous behaviour. Moreover, optogenetic inhibition and ablation of VLS D2-MSNs causes, respectively, transient and chronic reductions of goal-directed behaviours. Our data demonstrate that the circuitry containing VLS D2-MSNs control motivated behaviours and that VLS D2-MSN loss-of-function is a possible cause of motivation deficits in neurodegenerative diseases.

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Altered Tau Isoform Ratio Caused by Loss of FUS and SFPQ Function Leads to FTLD-like Phenotypes

Ishigaki Shinsuke, Fujioka Yusuke, Okada Yohei, Riku Yuichi, Udagawa Tsuyoshi, Honda Daiyu, Yokoi Satoshi, Endo Kuniyuki, Ikenaka Kensuke, Takagi Shinnosuke, Iguchi Yohei, Sahara Naruhiko, Takashima Akihiko, Okano Hideyuki, Yoshida Mari, Warita Hitoshi, Aoki Masashi, Watanabe Hirohisa, Okado Haruo, Katsuno Masahisa, Sobue Gen

Cell Reports 18 （ 5 ） 1118 - 1131 2017年01月

ISSN 2211-1247

Fused in sarcoma (FUS) and splicing factor, proline- and glutamine-rich (SFPQ) are RNA binding proteins that regulate RNA metabolism. We found that alternative splicing of the Mapt gene at exon 10, which generates 4-repeat tau (4R-T) and 3-repeat tau (3R-T), is regulated by interactions between FUS and SFPQ in the nuclei of neurons. Hippocampus-specific FUS- or SFPQ-knockdown mice exhibit frontotemporal lobar degeneration (FTLD)-like behaviors, reduced adult neurogenesis, accumulation of phosphorylated tau, and hippocampal atrophy with neuronal loss through an increased 4R-T/3R-T ratio. Normalization of this increased ratio by 4R-T-specific silencing results in recovery of the normal phenotype. These findings suggest a biological link among FUS/SFPQ, tau isoform alteration, and phenotypic expression, which may function in the early pathomechanism of FTLD.

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Efficient induction of dopaminergic neuron differentiation from induced pluripotent stem cells reveals impaired mitophagy in PARK2 neurons

Suzuki Sadafumi, Akamatsu Wado, Kisa Fumihiko, Sone Takefumi, Ishikawa Kei ichi, Kuzumaki Naoko, Katayama Hiroyuki, Miyawaki Atsushi, Hattori Nobutaka, Okano Hideyuki

Biochemical and Biophysical Research Communications 483 （ 1 ） 88 - 93 2017年01月

ISSN 0006-291X

Patient-specific induced pluripotent stem cells (iPSCs) show promise for use as tools for in vitro modeling of Parkinson's disease. We sought to improve the efficiency of dopaminergic (DA) neuron induction from iPSCs by the using surface markers expressed in DA progenitors to increase the significance of the phenotypic analysis. By sorting for a CD184^high/CD44^- fraction during neural differentiation, we obtained a population of cells that were enriched in DA neuron precursor cells and achieved higher differentiation efficiencies than those obtained through the same protocol without sorting. This high efficiency method of DA neuronal induction enabled reliable detection of reactive oxygen species (ROS) accumulation and vulnerable phenotypes in PARK2 iPSCs-derived DA neurons. We additionally established a quantitative system using the mt-mKeima reporter system to monitor mitophagy in which mitochondria fuse with lysosomes and, by combining this system with the method of DA neuronal induction described above, determined that mitophagy is impaired in PARK2 neurons. These findings suggest that the efficiency of DA neuron induction is important for the precise detection of cellular phenotypes in modeling Parkinson's disease.

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Sox21 deletion in mice causes postnatal growth deficiency without physiological disruption of hypothalamic-pituitary endocrine axes

Cheung Leonard Y M, Okano Hideyuki, Camper Sally A.

Molecular and Cellular Endocrinology 439 213 - 223 2017年01月

ISSN 0303-7207

The hypothalamic-pituitary axes are the coordinating centers for multiple endocrine gland functions and physiological processes. Defects in the hypothalamus or pituitary gland can cause reduced growth and severe short stature, affecting approximately 1 in 4000 children, and a large percentage of cases of pituitary hormone deficiencies do not have an identified genetic cause. SOX21 is a protein that regulates hair, neural, and trophoblast stem cell differentiation. Mice lacking Sox21 have reduced growth, but the etiology of this growth defect has not been described. We studied the expression of Sox21 in hypothalamic-pituitary development and examined multiple endocrine axes in these mice. We find no evidence of reduced intrauterine growth, food intake, or physical activity, but there is evidence for increased energy expenditure in mutants. In addition, despite changes in pituitary hormone expression, hypothalamic-pituitary axes appear to be functional. Therefore, SOX21 variants may be a cause of non-endocrine short stature in humans.

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Cochlear Cell Modeling Using Disease-Specific iPSCs Unveils a Degenerative Phenotype and Suggests Treatments for Congenital Progressive Hearing Loss

Hosoya Makoto, Fujioka Masato, Sone Takefumi, Okamoto Satoshi, Akamatsu Wado, Ukai Hideki, Ueda Hiroki R., Ogawa Kaoru, Matsunaga Tatsuo, Okano Hideyuki

Cell Reports 18 （ 1 ） 68 - 81 2017年01月

ISSN 2211-1247

Hearing impairments are the most common symptom of congenital defects, and they generally remain intractable to treatment. Pendred syndrome, the most frequent syndromic form of hereditary hearing loss, is associated with mutations in the anion exchanger pendrin. Loss of pendrin function as an anion exchanger is thought to be causative, but rodent models do not exhibit progressive deafness. Here, we report a degenerative phenotype exhibiting mutant pendrin aggregates and increased susceptibility to cellular stresses in cochlear epithelial cells induced from patient-derived induced pluripotent stem cells (iPSCs). These degenerative phenotypes were rescued by site-specific gene corrections. Moreover, low-dose rapamycin and metformin reduced aggregation and cell death. Our results provide an unexpected, comprehensive understanding of deafness due to “degenerative cochlear disease” and may contribute to rational therapeutic development. This iPSC-based disease model provides an approach to the study of pathogenesis and therapeutic development for hereditary hearing loss.

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Involvement of the septo-hippocampal cholinergic pathway in association with septal acetylcholinesterase upregulation in a mouse model of tauopathy

Hara Yuko, Motoi Yumiko, Hikishima Keigo, Mizuma Hiroshi, Onoe Hirotaka, Matsumoto Shin Ei, Elahi Montasir, Okano Hideyuki, Aoki Shigeki, Hattori Nobutaka

Current Alzheimer Research 14 （ 1 ） 94 - 103 2017年01月

ISSN 1567-2050

Background: Cholinergic cell loss in the basal forebrain, the major source of hippocampal cholinergic projections, has been implicated in Alzheimer’s disease. Objective: To examine whether the septohippocampal pathway is involved in tauopathy model mice and to elucidate the tau-associated mechanism underlying cholinergic alteration. Methods: Adult (6 to 8 months old) and old (16 to 18 months old) transgenic mice expressing wild-type human tau, Tg601, were examined using Ex vivo diffusion tensor magnetic resonance imaging (DTI) and 2-[¹⁸F]fluoro- 2-deoxy-D-glucose positron emission tomography (FDG-PET). Choline acetyltransferase (ChAT)-positive neurons in the medial septum (MS) were counted by stereological methods. Acetylcholinesterase (AChE) activity and AChE mRNA in 6 brain regions were measured. Results: Ex vivo DTI revealed that the number of fractional anisotropy (FA) streamlines in the septohippocampal tract decreased with age in Tg601 mice. The FA value in the septum was lower in old Tg601 mice than in non-tg mice. A voxel-based statistical analysis of FDG-PET revealed the presence of low glucose uptake areas, involving the MS in adults, and spread over regions including the hippocampal dentate gyrus in old mice. In the MS, the number of choline acetyltransferase (ChAT)-positive neurons decreased in old Tg601 mice. AChE activity and AChE mRNA T transcripts were exclusively higher in the septum. Conclusion: The upregulation of AChE in the septum may result in the selective degeneration of the septohippocampal cholinergic pathway in the tauopathy mouse model.

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The prospects of regenerative medicine combined with rehabilitative approaches for chronic spinal cord injury animal models

Tashiro Syoichi, Nakamura Masaya, Okano Hideyuki

Neural Regeneration Research 12 （ 1 ） 43 - 46 2017年01月

ISSN 1673-5374

Regenerative medicine has opened a window for functional recovery in acute-to-subacute phase spinal cord injury (SCI). By contrast, there are still only a few studies have focused on the treatment of the chronically injured spinal cord, in which cell-based regenerative medicine seems less effective. Since the majority of SCI patients are in the chronic phase, representing a major challenge for the clinical application of cell based regenerative medicine. Although combined therapies for the treatment of chronic SCI have attracted attention of researchers and its potential importance is also widely recognized, there had been very few studies involving rehabilitative treatments to date. In a recent study, we have demonstrated for the first time that treadmill training combined with cell transplantation significantly promotes functional recovery even in chronic SCI, not only in additive but also in synergistic manner. Even though we have succeeded to outline the profiles of recovery secondary to the combination therapy, the mechanism underlying the effects remain unsolved. In this review article, we summarize the present progress and consider the prospect of the cell-based regenerative medicine particularly combined with rehabilitative approaches for chronic SCI animal models.

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RNA-binding protein Qki5 regulates embryonic neural stem cells through pre-mRNA processing in cell adhesion signaling

Hayakawa-Yano Y, Suyama S, Nogami M, Yugami M, Koya I, Furukawa T, Zhou L,Abe M, Sakimura K, Takebayashi H, Nakanishi A, Okano H*, and Yano M.*

Genes & Dev 2017年

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Assessment of mitophagy in iPS cell-derived neurons

Ishikawa K., Yamaguchi A., Okano H., Akamatsu W.

Methods in Molecular Biology （Methods in Molecular Biology） 1759 59 - 67 2017年

ISSN 10643745

© 2017, Springer Science+Business Media New York. Aberrant mitochondrial function is associated with many neurological diseases. Mitophagy is a key mechanism for the elimination of damaged mitochondria and maintenance of mitochondrial homeostasis. Induced pluripotent stem (iPS) cell technologies developed over the last decade have allowed us to analyze functions of the human neuron. Here we describe an efficient induction method from human iPS cells to neurons, followed by an image-based mitophagy assay.

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STAT3: Down the R(h)oAd

Renault-Mihara F., Okano H.

Cytokine （Cytokine） 102 149 - 150 2017年

ISSN 10434666

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STAT3-regulated RhoA drives reactive astrocyte dynamics

Renault-Mihara F, Okano H

Cell Cycle 2017年

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iPSC-derived neural precursor cells: potential for cell transplantation therapy in spinal cord injury

Nagoshi N., Okano H.

Cellular and Molecular Life Sciences （Cellular and Molecular Life Sciences） 75 （ 6 ） 989 - 1000 2017年

ISSN 1420682X

© 2017, Springer International Publishing AG. A number of studies have demonstrated that transplantation of neural precursor cells (NPCs) promotes functional recovery after spinal cord injury (SCI). However, the NPCs had been mostly harvested from embryonic stem cells or fetal tissue, raising the ethical concern. Yamanaka and his colleagues established induced pluripotent stem cells (iPSCs) which could be generated from somatic cells, and this innovative development has made rapid progression in the field of SCI regeneration. We and other groups succeeded in producing NPCs from iPSCs, and demonstrated beneficial effects after transplantation for animal models of SCI. In particular, efficacy of human iPSC–NPCs in non-human primate SCI models fostered momentum of clinical application for SCI patients. At the same time, however, artificial induction methods in iPSC technology created alternative issues including genetic and epigenetic abnormalities, and tumorigenicity after transplantation. To overcome these problems, it is critically important to select origins of somatic cells, use integration-free system during transfection of reprogramming factors, and thoroughly investigate the characteristics of iPSC–NPCs with respect to quality management. Moreover, since most of the previous studies have focused on subacute phase of SCI, establishment of effective NPC transplantation should be evaluated for chronic phase hereafter. Our group is currently preparing clinical-grade human iPSC–NPCs, and will move forward toward clinical study for subacute SCI patients soon in the near future.

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Low immunogenicity of mouse induced pluripotent stem cell-derived neural stem cells

Itakura G, Ozaki M, Nagoshi N, Kawabata S, Nishiyama Y, Sugai K, Iida T, Rei Kashiwagi R, Ookubo T, Yastake K, Matsubayashi K, Kohyama J, Iwanami A, Matsumoto M, Nakamura M, and Okano H

Sci Rep 7 12996 2017年

RNA Sequencing Analysis Reveals Interactions between Breast Cancer or Melanoma Cells and the Tissue Microenvironment during Brain Metastasis

Sato Ryo, Nakano Teppei, Hosonaga Mari, Sampetrean Oltea, Harigai Ritsuko, Sasaki Takashi, Koya Ikuko, Okano Hideyuki, Kudoh Jun, Saya Hideyuki, Arima Yoshimi

BioMed Research International 2017 2017年

ISSN 2314-6133

Metastasis is the main cause of treatment failure and death in cancer patients. Metastasis of tumor cells to the brain occurs frequently in individuals with breast cancer, non-small cell lung cancer, or melanoma. Despite recent advances in our understanding of the causes and in the treatment of primary tumors, the biological and molecular mechanisms underlying the metastasis of cancer cells to the brain have remained unclear. Metastasizing cancer cells interact with their microenvironment in the brain to establish metastases. We have now developed mouse models of brain metastasis based on intracardiac injection of human breast cancer or melanoma cell lines, and we have performed RNA sequencing analysis to identify genes in mouse brain tissue and the human cancer cells whose expression is associated specifically with metastasis. We found that the expressions of the mouse genes Tph2, Sspo, Ptprq, and Pole as well as those of the human genes CXCR4, PLLP, TNFSF4, VCAM1, SLC8A2, and SLC7A11 were upregulated in brain tissue harboring metastases. Further characterization of such genes that contribute to the establishment of brain metastases may provide a basis for the development of new therapeutic strategies and consequent improvement in the prognosis of cancer patients.

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Plasticity and regeneration in the injured spinal cord after cell transplantation therapy

Nori S., Nakamura M., Okano H.

Progress in Brain Research 2017年

ISSN 0079-6123

Spinal cord injury (SCI) typically damages the long axonal tracts of the spinal cord which results in permanent disability. However, regeneration of the injured spinal cord is approaching reality according to the advances in stem cell biology. Cell transplantation therapy holds potential to lead to recovery following SCI through some positive mechanisms. Grafted cells induce plasticity and regeneration in the injured spinal cord by promoting remyelination of damaged axons, reconstruction of neural circuits by synapse formation between host neurons and graft-derived neurons, and secreting neurotrophic factors to promote axonal elongation as well as reduce retrograde axonal degeneration. In this review, we will delineate (1) the microenvironment of the injured spinal cord that influence the plasticity and regeneration capacity after SCI, (2) a number of different kinds of cell transplantation therapies for SCI that has been extensively studied by researchers, and (3) potential mechanisms of grafted cell-induced regeneration and plasticity in the injured spinal cord.

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Brief exposure to small molecules allows induction of mouse embryonic fibroblasts into neural crest-like precursors

Takayama Yuzo, Wakabayashi Tamami, Kushige Hiroko, Saito Yutaka, Shibuya Yoichiro, Shibata Shinsuke, Akamatsu Wado, Okano Hideyuki, Kida Yasuyuki S.

FEBS Letters 2017年

ISSN 0014-5793

In this study, we propose a novel method for inducing neuronal cells by briefly exposing them to small-molecule cocktails in a step-by-step manner. Global gene expression analysis with immunohistochemical staining and calcium flux assays reveal the generation of neurons from mouse embryonic fibroblasts. In addition, time-lapse imaging of neural precursor-specific enhancer expression and global gene expression analyses show that the neurons are generated by passing through a neural crest-like precursor stage. Consistent with these results, the neural crest-like cells are able to differentiate into neural crest lineage cells, such as sympathetic neurons, adipocytes, osteocytes, and smooth muscle cells. Therefore, these results indicate that brief exposure to chemical compounds could expand and induce a substantial multipotent cell population without viral transduction.

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Differential X chromosome inactivation patterns during the propagation of human induced pluripotent stem cells

Andoh-Noda Tomoko, Akamatsu Wado, Miyake Kunio, Kobayashi Tetsuro, Ohyama Manabu, Kurosawa Hiroshi, Kubota Takeo, Okano Hideyuki

Keio Journal of Medicine 66 （ 1 ） 1 - 8 2017年

ISSN 0022-9717

Human induced pluripotent stem cells (hiPSCs) represent a potentially useful tool for studying te molecular mechanisms of disease thanks to their ability to generate patient-specific hiPSC clones. However, previous studies have reported that DNA methylation profiles, including those for imprinted genes, may change during passaging of hiPSCs. This is particularly problematic for hiPSC models of X-linked disease, because unstable X chromosome inactivation status may affect the detection of phenotypes. In the present study, we examined the epigenetic status of hiPSCs derived from patients with Rett syndrome, an X-linked disease, during long-term culture. To analyze X chromosome inactivation, we used a methylation-specific polymerase chain reaction (MSP) to assay the human androgen receptor locus (HUMARA). We found that single cell-derived hiPSC clones exhibit various states of X chromosome inactivation immediately after clonal isolation, even when established simultaneously from a single donor. X chromosome inactivation states remain variable in hiPSC clones at early passages, and this variability may affect cellular phenotypes characteristic of X-linked diseases. Careful evaluation of X chromosome inactivation in hiPSC clones, particularly in early passages, by methods such as HUMARA-MSP, is therefore important when using patient-specific hiPSCs to model X-linked disease.

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Whole-Genome DNA Methylation Analyses Revealed Epigenetic Instability in Tumorigenic Human iPS Cell-Derived Neural Stem/Progenitor Cells

Iida Tsuyoshi, Iwanami Akio, Sanosaka Tsukasa, Kohyama Jun, Miyoshi Hiroyuki, Nagoshi Narihito, Kashiwagi Rei, Toyama Yoshiaki, Matsumoto Morio, Nakamura Masaya, Okano Hideyuki

Stem Cells 2017年

ISSN 1066-5099

Although human induced pluripotent stem cell (hiPSC) derivatives are considered promising cellular resources for regenerative medicine, their tumorigenicity potentially limits their clinical application in hiPSC technologies. We previously demonstrated that oncogenic hiPSC-derived neural stem/progenitor cells (hiPSC-NS/PCs) produced tumor-like tissues that were distinct from teratomas. To gain insight into the mechanisms underlying the regulation of tumorigenicity in hiPSC-NS/PCs, we performed an integrated analysis using the Infinium HumanMethylation450 BeadChip array and the HumanHT-12 v4.0 Expression BeadChip array to compare the comprehensive DNA methylation and gene expression profiles of tumorigenic hiPSC-NS/PCs (253G1-NS/PCs) and non-tumorigenic cells (201B7-NS/PCs). Although the DNA methylation profiles of 253G1-hiPSCs and 201B7-hiPSCs were similar regardless of passage number, the methylation status of the global DNA methylation profiles of 253G1-NS/PCs and 201B7-NS/PCs differed; the genomic regions surrounding the transcriptional start site of the CAT and PSMD5 genes were hypermethylated in 253G1-NS/PCs but not in 201B7-NS/PCs. Interestingly, the aberrant DNA methylation profile was more pronounced in 253G1-NS/PCs that had been passaged more than 15 times. In addition, we identified aberrations in DNA methylation at the RBP1 gene locus; the DNA methylation frequency in RBP1 changed as 253G1-NS/PCs were sequentially passaged. These results indicate that different NS/PC clones have different DNA methylomes and that DNA methylation patterns are unstable as cells are passaged. Therefore, DNA methylation profiles should be included in the criteria used to evaluate the tumorigenicity of hiPSC-NS/PCs in the clinical setting.

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Changeability of the fully methylated status of the 15q11.2 region in induced pluripotent stem cells derived from a patient with Prader-Willi syndrome

Okuno Hironobu, Nakabayashi Kazuhiko, Abe Kousei, Ando Takayuki, Sanosaka Tsukasa, Kohyama Jun, Akamatsu Wado, Ohyama Manabu, Takahashi Takao, Kosaki Kenjiro, Okano Hideyuki

Congenital Anomalies 2017年

ISSN 0914-3505

Prader-Will syndrome (PWS) is characterized by hyperphagia, growth hormone deficiency and central hypogonadism caused by the dysfunction of the hypothalamus. Patients with PWS present with methylation abnormalities of the PWS-imprinting control region in chromosome 15q11.2, subject to parent-of-origin-specific methylation and controlling the parent-of-origin-specific expression of other paternally expressed genes flanking the region. In theory, the reversal of hypermethylation in the hypothalamic cells could be a promising strategy for the treatment of PWS patients, since cardinal symptoms of PWS patients are correlated with dysfunction of the hypothalamus. The genome-wide methylation status dramatically changes during the reprograming of somatic cells into induced pluripotent stem cells (iPSCs) and during the in vitro culture of iPSCs. Here, we tested the methylation status of the chromosome 15q11.2 region in iPSCs from a PWS patient using pyrosequencing and a more detailed method of genome-wide DNA methylation profiling to reveal whether iPSCs with a partially unmethylated status for the chromosome 15q11.2 region exhibit global methylation aberrations. As a result, we were able to show that a fully methylated status for chromosome 15q11.2 in a PWS patient could be reversed to a partially unmethylated status in at least some of the PWS-iPSC lines. Genome-wide DNA methylation profiling revealed that the partial unmethylation occurred at differentially methylated regions located in chromosome 15q11.2, but not at other differentially methylated regions associated with genome imprinting. The present data potentially opens a door to cell-based therapy for PWS patients and, possibly, patients with other disorders associated with genomic imprinting.

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Applications of induced pluripotent stem cell technologies in spinal cord injury

Nagoshi Narihito, Okano Hideyuki

Journal of Neurochemistry 2017年

ISSN 0022-3042

Numerous basic research studies have suggested the potential efficacy of neural precursor cell (NPC) transplantation in spinal cord injury (SCI). However, in most such studies, the origin of the cells used was mainly fetal tissue or embryonic stem cells, both of which carry potential ethical concerns with respect to clinical use. The development of induced pluripotent stem cells (iPSCs) opened a new path toward regenerative medicine for SCI. iPSCs can be generated from somatic cells by induction of transcription factors, and induced to differentiate into NPCs with characteristics of cells of the central nervous system. The beneficial effect of iPSC-derived NPC transplantation has been reported from our group and others working in rodent and non-human primate models. These promising results facilitate the application of iPSCs for clinical applications in SCI patients. However, iPSCs also have issues, such as genetic/epigenetic abnormalities and tumorigenesis because of the artificial induction method, that must be addressed prior to clinical use. The selection of somatic cells, generation of integration-free iPSCs, and characterization of differentiated NPCs with thorough quality management are all needed to address these potential risks. To enhance the efficacy of the transplanted iPSC-NPCs, especially at chronic phase of SCI, administration of a chondroitinase or semaphorin3A inhibitor represents a potentially important means of promoting axonal regeneration through the lesion site. The combined use of rehabilitation with such cell therapy approaches is also important, as repetitive training enhances neurite outgrowth of transplanted cells and strengthens neural circuits at central pattern generators. Our group has already evaluated clinical grade iPSC-derived NPCs, and we look forward to initiating clinical testing as the next step toward determining whether this approach is safe and effective for clinical use. This article is part of the series "60th Anniversary of the Japanese Society for Neurochemistry".

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Regulation of RhoA by STAT3 coordinates glial scar formation

Renault-Mihara Francois, Mukaino Masahiko, Shinozaki Munehisa, Kumamaru Hiromi, Kawase Satoshi, Baudoux Matthieu, Ishibashi Toshiki, Kawabata Soya, Nishiyama Yuichiro, Sugai Keiko, Yasutake Kaori, Okada Seiji, Nakamura Masaya, Okano Hideyuki

Journal of Cell Biology 216 （ 8 ） 2533 - 2550 2017年

ISSN 0021-9525

Understanding how the transcription factor signal transducer and activator of transcription-3 (STAT3) controls glial scar formation may have important clinical implications. We show that astrocytic STAT3 is associated with greater amounts of secreted MMP2, a crucial protease in scar formation. Moreover, we report that STAT3 inhibits the small GTPase RhoA and thereby controls actomyosin tonus, adhesion turnover, and migration of reactive astrocytes, as well as corralling of leukocytes in vitro. The inhibition of RhoA by STAT3 involves ezrin, the phosphorylation of which is reduced in STAT3-CKO astrocytes. Reduction of phosphatase and tensin homologue (PTEN) levels in STAT3-CKO rescues reactive astrocytes dynamics in vitro. By specific targeting of lesion-proximal, reactive astrocytes in Nestin-Cre mice, we show that reduction of PTEN rescues glial scar formation in Nestin-Stat3^+/- mice. These findings reveal novel intracellular signaling mechanisms underlying the contribution of reactive astrocyte dynamics to glial scar formation.

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Three-dimensional kinematic and kinetic analysis of quadrupedal walking in the common marmoset (Callithrix jacchus)

Shimada Hikaru, Kanai Ryogo, Kondo Takahiro, Yoshino-Saito Kimika, Uchida Akito, Nakamura Masaya, Ushiba Junichi, Okano Hideyuki, Ogihara Naomichi

Neuroscience Research 2017年

ISSN 0168-0102

The common marmoset has recently gained a great deal of attention as an experimental primate model for biological science and medical research. To use the common marmoset for development of novel treatments and rehabilitation for locomotor disorders, it is crucial to understand fundamental baseline characteristics of locomotion in this species. Therefore, in the present study we performed kinematic and kinetic analyses of quadrupedal locomotion in this animal. A total of 14 common marmosets walking quadrupedally along a walkway were analyzed using synchronized high-speed cameras, with two force platforms set in the walkway. Our results demonstrated that the marmoset uses a lateral sequence walking pattern, in contrast to the macaque and other primates, which usually adopt a diagonal sequence pattern. Furthermore, peak vertical ground reaction force on the forelimb was larger than that on the hindlimb. The rate of energy recovery for quadrupedal walking in the common marmoset was much smaller than that in the macaque, indicating that the marmoset generally utilizes bouncing mechanics in locomotion, even though the duty factor is >. 0.5. This description of locomotor characteristics of intact marmosets may serve as a basis for comparative analyses of changes in gait due to rehabilitation and regenerative treatments.

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Neuroscience research using non-human primate models and genome editing

Kishi Noriyuki, Okano Hideyuki

Genome Editing in Neurosciences 73 - 81 2017年

The common marmoset (Callithrix jacchus) is a small New World non-human primate indigenous to northeastern Brazil. This species has been attracting the attention of biomedical researchers and neuroscientists for its ease of handling and colony maintenance, unique behavioral characteristics, and several human-like traits, such as enriched social vocal communication and strong relationships between parents and offspring. Its high reproductive efficiency makes it particularly amenable for use in the development of transgenic and genome editing technologies in a non-human primate model. Our group has recently generated transgenic marmosets with germ line transmission, opening new avenues in primate research. In this chapter, we describe recent advances in neuroscience and disease research using common marmosets, and we outline potential uses of genome editing in non-human primates toward the development of knock-in/knock-out marmosets.

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Astrocyte-mediated infantile-onset leukoencephalopathy mouse model

583) Sugio S, Tohyama K, Oku S, Fujiyoshi K, Yoshimura T, Hikishima K, Yano R, Fukuda T, Nakamura M, Okano H, Watanabe M, Fukata M, Ikenaka K, F Tanaka K

GLIA 65(1) 150 - 168 2017年

Technical feasibility of visualizing myenteric plexus using confocal laser endomicroscopy

591) Kobayashi M, Sumiyama K, Shimojima N, Shimojima N, Ieiri S, Okano H, Kamba S, Fujimura T, Hirobe S, Kuroda T, Takahashi-Fujigasaki J

J Gastroenterol Hepatol 2017年

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Enhanced functional recovery from spinal cord injury in aged mice after stem cell transplantation through HGF induction

593) Takano M, Kawabata S, Shibata S, Yasuda A, Nori S, Tsuji O, Iwanami A, Kohyama J, Ebise H, Horiuchi K, Okano H*, Nakamura M*

Stem Cell Reports 2017年

In vivo microscopic voxel-based morphometry with a brain template to characterize strain-specific structures in the mouse brain

Hikishima K, Komaki Y, Seki F, Ohnishi Y, Okano HJ, Okano H

Sci Rep 7(1) 85 2017年

Generation of neural cells using iPSCs from sleep bruxism patients with 5-HT2A polymorphism

Hoashi Y, Okamoto S, Abe Y, Matsumoto T, Tanaka J, Yoshida Y, Imaizumi K, Mishima K, Akamatsu W, Okano H, Baba K

J Prosthodont Res 61(3) 242 - 250 2017年

Structural insight into recognition of r(UAG) by Musashi-1 RBD2, and construction of a model of Musashi-1 RBD1-2 bound to the minimum target RNA

Ryo Iwaoka, Takashi Nagata , Kengo Tsuda, Takao Imai, Hideyuki Okano, Naohiro Kobayashi, Masato Katahira

Molecules 22 (7) 2017年

Backbone and side chain assignments of the second RNA-binding domain of Musashi-1in its free form and in complex with 5-mer RNA

Iwaoka R, Nagata T, Tsuda K, Imai T, Okano H, Kobayashi N, Katahira M

Biomol NMR Assign 2017年

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Mapping orbitofrontal-limbic maturation in non-human primates: A longitudinal magnetic resonance imaging study

Uematsu A, Hata J, Komaki Y, Seki F, Yamada C, Okahara N, Kurotaki Y, Sasaki E, Okano H

Neuroimage 163 55 - 67 2017年

Physiological effects of a habituation procedure for functional MRI in awake mice using a cryogenic radiofrequency probe.

岡野栄之

Journal of Neuroscience Methods 1 （ 274 ） 38 - 48 2016年10月

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Pathological classification of human iPSC-derived neural stem/progenitor cells towards safety assessment of transplantation therapy for CNS diseases.

岡野栄之

Molecular Brain 2016年09月

共著

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Sox21 deletion in mice causes postnatal growth deficiency without physiological disruption of hypothalamic-pituitary endocrine axes.

岡野栄之

Molecular and Cellular Endocrinology 2016年09月

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Pretreatment with a γ-Secretase Inhibitor Prevents Tumor-like Overgrowth in Human iPSC-Derived Transplants for Spinal Cord Injury.

岡野栄之

Stem Cell Reports 2016年09月

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Identification of hepta-histidine as a candidate drug for Huntington's disease by in silico-in vitro- in vivo-integrated screens of chemical libraries.

岡野栄之

SCIENTIFIC REPORTS 22 （ 6 ） 33861 2016年09月

共著

JMJD1C Exhibits Multiple Functions in Epigenetic Regulation during Spermatogenesis.

岡野栄之

PLoS One 20 （ 11(9) ） e0163466 2016年09月

共著

Worldwide initiatives to advance brain research.

岡野栄之

Nature Neuroscience 26 （ 19(9) ） 1118 - 1122 2016年08月

共著

Neural changes in the primate brain correlated with the evolution of complex motor skills.

岡野栄之

SCIENTIFIC REPORTS 8 （ 6 ） 31084 2016年08月

研究論文（学術雑誌）, 共著

Combined treatment with chondroitinase ABC and treadmill rehabilitation for chronic severe spinal cord injury in adult rats.

岡野栄之

Neuroscience Research 2016年08月

研究論文（学術雑誌）, 共著

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Functional Recovery from Neural Stem/Progenitor Cell Transplantation Combined with Treadmill Training in Mice with Chronic Spinal Cord Injury.

岡野栄之

SCIENTIFIC REPORTS 3 （ 6 ） 30898 2016年08月

研究論文（学術雑誌）, 共著

Expression pattern of wolframin, the WFS1 (Wolfram syndrome-1 gene) product, in common marmoset (Callithrix jacchus) cochlea.

岡野栄之

Neuroreport 3 （ 27(11) ） 833 - 836 2016年08月

研究論文（学術雑誌）

SAMD9 mutations cause a novel multisystem disorder, MIRAGE syndrome, and are associated with loss of chromosome 7.

岡野栄之

nature genetics 48(7) 792 - 797 2016年07月

研究論文（学術雑誌）

Induction of artificial cancer stem cells from tongue cancer cells by defined reprogramming factors.

岡野栄之

BioMedCentral 27 （ 16 ） 548 2016年07月

研究論文（学術雑誌）, 共著

In vitro characterization of neurite extension using induced pluripotent stem cells derived from lissencephaly patients with TUBA1A missense mutations.

岡野栄之

Molecular Brain 19 （ 9(1) ） 70 2016年07月

研究論文（学術雑誌）, 共著

Generation of a Nonhuman Primate Model of Severe Combined Immunodeficiency Using Highly Efficient Genome Editing.

岡野栄之

Cell Stem Cell 7 （ 19(1) ） 127 - 138 2016年07月

研究論文（学術雑誌）, 共著

Safe and efficient method for cryopreservation of human induced pluripotent stem cell-derived neural stem and progenitor cells by a programmed freezer with a magnetic field.

岡野栄之

Neuroscience Research 107 20 - 29 2016年06月

研究論文（学術雑誌）

Distinct Expression Pattern of a Deafness Gene, KIAA1199, in a Primate Cochlea.

岡野栄之

BioMedReserch International 2016年06月

研究論文（学術雑誌）, 共著

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Involvement of the septo-hippocampal cholinergic pathway in association with septal acetylcholinesterase upregulation in a mouse model of tauopathy.

岡野栄之

Curr Alzheimer Res 2016年06月

研究論文（学術雑誌）

Image-based detection and targeting of therapy resistance in pancreatic adenocarcinoma.

岡野栄之

Nature 6 （ 534(7607) ） 407 - 411 2016年06月

研究論文（学術雑誌）, 共著

H1foo Has a Pivotal Role in Qualifying Induced Pluripotent Stem Cells.

岡野栄之

Stem Cell Reports 14 （ 6(6) ） 825 - 833 2016年06月

研究論文（学術雑誌）, 共著

MIF Maintains the Tumorigenic Capacity of Brain Tumor-Initiating Cells by Directly Inhibiting p53.

岡野栄之

Cancer Reserch 1 （ 76(9) ） 2813 - 2823 2016年05月

研究論文（学術雑誌）, 共著

LNGFR+THY-1+ human pluripotent stem cell-derived neural crest-like cells have the potential to develop into mesenchymal stem cells.

岡野栄之

Differentiation 2016年05月

研究論文（学術雑誌）

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Tumour resistance in induced pluripotent stem cells derived from naked mole-rats.

岡野栄之

nature communications 10 （ 7 ） 11471 2016年05月

研究論文（学術雑誌）, 共著

Musashi mediates translational repression of the Drosophila hypoxia inducible factor.

岡野栄之

Nucleic Acids Research 2016年05月

研究論文（学術雑誌）, 共著

Establishment of In Vitro FUS-Associated Familial Amyotrophic Lateral Sclerosis Model Using Human Induced Pluripotent Stem Cells.

岡野栄之

Stem Cell Reports 12 （ 6(4) ） 496 - 510 2016年04月

研究論文（学術雑誌）, 共著

Establishment of Human Neural Progenitor Cells from Human Induced Pluripotent Stem Cells with Diverse Tissue Origins.

岡野栄之

Stem Cells International 2016年04月

研究論文（学術雑誌）, 共著

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Sustained Effect of Hyaluronic Acid in Subcutaneous Administration to the Cochlear Spiral Ganglion.

岡野栄之

PLOS ONE 21 （ 11(4) ） e0153957 2016年04月

研究論文（学術雑誌）, 共著

Overlapping expression of anion exchangers in the cochlea of a non-human primate suggests functional compensation.

岡野栄之

Neuroscience Reserch 2016年04月

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Robust production of human neural cells by establishing neuroepithelial-like stem cells from peripheral blood mononuclear cell-derived feeder-free iPSCs under xeno-free conditions.

岡野栄之

Neuroscience Reserch 2016年04月

研究論文（学術雑誌）, 共著

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Functional Neurons Generated from T Cell-Derived Induced Pluripotent Stem Cells for Neurological Disease Modeling.

岡野栄之

Stem Cell Reports 8 （ 6(3) ） 422 - 435 2016年03月

研究論文（学術雑誌）, 共著

Application of q-Space Diffusion MRI for the Visualization of White Matter.

岡野栄之

THE JOURNAL OF NEUROSCIENCE 2 （ 36(9) ） 2796 - 2808 2016年03月

研究論文（学術雑誌）, 共著

Fluorescence Visualization of the Enteric Nervous Network in a Chemically Induced Aganglionosis Model.

岡野栄之

PLOS ONE 4 （ 11(3) ） e0150579 2016年03月

研究論文（学術雑誌）, 共著

Distinct Expression Patterns Of Causative Genes Responsible For Hereditary Progressive Hearing Loss In Non-Human Primate Cochlea.

岡野栄之

SCIENTIFIC REPORTS 2016年02月

研究論文（学術雑誌）, 共著

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Applications of Mesenchymal Stem Cells and Neural Crest Cells in Craniofacial Skeletal Research.

岡野栄之

StemCells International 2016年02月

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RNA regulation went wrong in neurodevelopmental disorders: The example of Msi/Elavl RNA binding proteins.

岡野栄之

International Journal of Developmental Neuroscience 2016年01月

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Erratum:Investigation of the fatty acid transporter-encoding genes SLC27A3 and SLC27A4 in autism.

岡野栄之

SCIENTIFIS REPORTS 29 （ 6 ） 20268 2016年01月

研究論文（学術雑誌）, 共著

MHC-compatible bone marrow stromal/stem cells trigger fibrosis by activating host T cells in a scleroderma mouse model.

岡野栄之

Elife 26 （ 5 ） e09394 2016年01月

研究論文（学術雑誌）, 共著

To the Cloud! A Grassroots Proposal to Accelerate Brain Science Discovery

Vogelstein JT, Mensh B, Häusser M, Spruston N, Evans AC, Kording K, Amunts K, Ebell C, Muller J, Telefont M, Hill S, Koushika SP, Calì C, Valdés-Sosa PA, Littlewood PB, Koch C, Saalfeld S, Kepecs A, Peng H, Halchenko YO, Kiar G, Poo MM, Poline JB, Milham MP, Schaffer AP, Gidron R, Okano H, Calhoun VD, Chun M, Kleissas DM, Vogelstein RJ, Perlman E, Burns R, Huganir R, Miller MI

Neuron 92(3) 622 - 627 2016年

Modeling Rett Syndrome Using Human Induced Pluripotent Stem Cells.

岡野栄之

CNS & Neurological Disorders - Drug Targets 15(5) 544 - 550 2016年

研究論文（学術雑誌）, 共著

Induction of artificial cancer stem cells from tongue cancer cells by defined reprogramming factors

567) Harada K, Ferdous T, Cui D, Kuramitsu Y, Matsumoto T, Ikeda E, Okano H, Ueyama Y

BMC Cancer 16(1) 548 2016年

Analysis of induced pluripotent stem cells carrying 22q11.2 deletion

569) Toyoshima M, Akamatsu W, Okada Y, Ohnishi T, Balan S, Hisano Y, Iwayama Y, Toyota T, Matsumoto T, Itasaka N, Sugiyama S, Tanaka M, Yano M, Dean B, Okano H, Yoshikawa T

Translational Psychiatry 6(11) e934 2016年

Modeling neurological diseases with induced pluripotent cells reprogrammed from immortalized lymphoblastoid cell lines

573) Fujimori K, Tezuka T, Ishiura H, Mitsui J, Doi K, Yoshimura J, Tada H, Matsumoto T, Isoda M, Hashimoto R, Hattori N, Takahashi T, Morishita S, Tsuji S, Akamatsu W, Okano H

Mol Brain 9(1) 88 2016年

Functional brain mapping using specific sensory-circuit stimulation and a theoretical graph network analysis in mice with neuropathic allodynia

575) Komaki Y, Hikishima K, Shibata S, Konomi T, Seki F, Yamada M, Miyasaka N, Fujiyoshi K, Okano HJ, Nakamura M, Okano H.

Sci Rep 6 37802 2016年

Notch2 signaling regulates the proliferation of murine bone marrow-derived mesenchymal stem/stromal cells via c-Myc expression

576) Sato Y, Mabuchi Y, Miyamoto K, Araki D, Niibe K, Houlihan DD, Morikawa S, Nakagawa T, Nakajima T, Akazawa C, Hori S, Okano H, Matsuzaki Y

PLoS One 11(11) e0165946 2016年

CHD7 promotes proliferation of neural stem cells mediated by MIF

577) Ohta S, Yaguchi T, Chneiweiss H, Okuno H, Kawakami Y, Okano H

Molecular Brain 9(1) 96 2016年

Changeability of the fully methylated status of the 15q11.2 region in induced pluripotent stem cells derived from a patient with Prader-Willi syndrome

578) Okuno H, Nakabayashi K, Abe K, Ando T, Sanosaka T, Kohyama J, Akamatsu W, Ohyama M, Takahashi T, Kosaki K, Okano H

Congenit Anom 18(1) 68 - 81 2016年

Functional comparison of neuronal cells differentiated from human induced pluripotent stem cell–derived neural stem cells under different oxygen and medium conditions

579) Yamazaki K, Fukushima K, Sugawara M, Tabata Y, Imaizumi Y, Ishihara Y, Ito M, Tsukahara K, Kohyama J, Okano H

Journal of Biomolecular Screening 21(10) 1054 - 1064 2016年

Heterochronic microRNAs in temporal specification of neural stem cells: application toward rejuvenation

Shimazaki T, Okano H

Aging and Mechanism of Disease 2 15014 2016年

A new era for Inflammation and Regeneration

Okano H

Inflammation and Regeneration 36 3 2016年

How can brain mapping initiatives cooperate to achieve the same goal?

Okano H, Yamamori T

Nat Rev Neurosci 17 733 - 734 2016年

Brain/MINDS: a Japanese National Brain Project for Marmoset Neuroscience

Okano H, Sasaki E, Yamamori T, Iriki A, Shimogori T, Yamaguchi Y, Kasai K and Miyawaki A

Neuron 92 (3) 582 - 590 2016年

Rapid, efficient, and simple motor neuron differentiation from human pluripotent stem cells.

岡野栄之

Moleculer Brain 1 （ 8(1) ） 79 2015年12月

研究論文（学術雑誌）, 共著

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The Mediator Kinase Module Restrains Epidermal Growth Factor Receptor Signaling and Represses Vulval Cell Fate Specification in Caenorhabditis elegans.

岡野栄之

Genetics 202(2) 583 - 599 2015年12月

研究論文（学術雑誌）, 共著

Grafted Human iPS Cell-Derived Oligodendrocyte Precursor Cells Contribute to Robust Remyelination of Demyelinated Axons after Spinal Cord Injury

岡野栄之

Stem Cell Reports 12 （ 6(1) ） 1 - 8 2015年12月

研究論文（学術雑誌）, 共著

Sphere-formation culture of testicular germ cells in the common marmoset, a small New World monkey.

岡野栄之

Primates 57(1) 129 - 135 2015年11月

研究論文（学術雑誌）, 共著

Analysis of RNA metabolism in peripheral WBCs of TDP-43 KI mice identifies novel biomarkers of ALS.

岡野栄之

Neuroscience Reserch 106 12 - 22 2015年11月

研究論文（学術雑誌）, 共著

α-synuclein aggregation in the olfactory bulb of middle-aged common marmoset.

岡野栄之

Neuroscience Reserch 106 55 - 61 2015年11月

研究論文（学術雑誌）

Resequencing of the common marmoset genome improves genome assemblies and gene-coding sequence analysis.

岡野栄之

Science Report 20 （ 5 ） 16894 2015年11月

研究論文（学術雑誌）, 共著

Controlling the Regional Identity of hPSC-Derived Neurons to Uncover Neuronal Subtype Specificity of Neurological Disease Phenotypes.

岡野栄之

Stem Cell Reports 8 （ 5(6) ） 1010 - 1022 2015年11月

研究論文（学術雑誌）

Investigation of the fatty acid transporter-encoding genes SLC27A3 and SLC27A4 in autism.

岡野栄之

Science Report 29 （ 6 ） 20268 2015年11月

共著

Muscle Satellite Cell Protein Teneurin-4 Regulates Differentiation During Muscle Regeneration.

岡野栄之

STEM CELLS 33(10) 3017 - 3027 2015年10月

研究論文（学術雑誌）, 共著

Gastric carcinoma: evaluation with diffusion-tensor MR imaging and tractography ex vivo.

岡野栄之

Magnetic Resonance Imaging 34(2) 144 - 151 2015年10月

研究論文（学術雑誌）

Purified Human Dental Pulp Stem Cells Promote Osteogenic Regeneration.

岡野栄之

Journal of Dental Reserch 95(2) 206 - 214 2015年10月

研究論文（学術雑誌）, 共著

IL-10 gene transfer upregulates arcuate POMC and ameliorates hyperphagia, obesity and diabetes by substituting for leptin.

岡野栄之

International Journal of Obesity 40(3) 425 - 433 2015年10月

研究論文（学術雑誌）, 共著

Novel in vivo imaging analysis of an inner ear drug delivery system: drug availability in inner ear following different dose of systemic drug injections.

岡野栄之

Hearing Reserch 330(Pt A) 142 - 146 2015年09月

研究論文（学術雑誌）, 共著

Neural Stem/Progenitor Cells Promote Functional Recovery through Synapse Reorganization with Spared Host Neurons after Spinal Cord Injury.

岡野栄之

Stem Cell Reports 11 （ 5(2) ） 264 - 277 2015年08月

研究論文（学術雑誌）

Endometrial Side Population Cells: Potential Adult Stem/Progenitor Cells in Endometrium.

岡野栄之

Biol Reprod 93 （ (4) ） 84 2015年08月

研究論文（学術雑誌）

STAT3-dependent reactive astrogliosis in the spinal dorsal horn underlies chronic itch.

岡野栄之

nature medicine 21(8) 927 - 931 2015年08月

研究論文（学術雑誌）

MicroRNA-153 Regulates the Acquisition of Gliogenic Competence by Neural Stem Cells.

岡野栄之

Stem Cell Reports 8 （ 5(3) ） 365 - 377 2015年07月

研究論文（学術雑誌）

Human somatic stem cell-based therapy for cartilage regeneration.

岡野栄之

ANNALS OF TRANSLATIONAL MEDISINE 3(Suppl 1) S17 2015年05月

研究論文（学術雑誌）, 共著

Immuno-Electron Microscopy and Electron Microscopic In Situ Hybridization for Visualizing piRNA Biogenesis Bodies in Drosophila Ovaries.

岡野栄之

Methods in Molecular Biology 1328 163 - 178 2015年

研究論文（学術雑誌）

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