Papers - Hasuwa, Hidetoshi

GPAT2 is required for piRNA biogenesis, transposon silencing, and maintenance of spermatogonia in mice†

Shiromoto Y., Kuramochi-Miyagawa S., Nagamori I., Chuma S., Arakawa T., Nishimura T., Hasuwa H., Tachibana T., Ikawa M., Nakano T.

Biology of reproduction (Biology of reproduction)  101 ( 1 ) 248 - 256 2019.07

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© The Author(s) 2019. Published by Oxford University Press on behalf of Society for the Study of Reproduction. PIWI-interacting RNAs (piRNAs), a subclass of germ cell-specific noncoding small RNAs, are essential for de novo DNA methylation of retrotransposon genes in embryonic testes. PIWIL2/MILI, one of three mouse PIWI family members, is indispensable for piRNA production, DNA methylation of retrotransposons presumably via piRNA, and normal spermatogenesis. In vitro analysis using germline stem cells (GS cells) revealed that glycerol-3-phosphate acyltransferase 2 (GPAT2), which is a mitochondrial outer membrane protein involved in generation of lysophosphatidic acid (LPA) and highly expressed in testes, plays important roles in spermatogenesis. Namely, GPAT2 binds to PIWIL2 and is closely involved in the biogenesis of piRNAs; this process is independent of its enzymatic activity on LPA. However, GS cells recapitulate only a limited phase of spermatogenesis and the biological functions of GPAT2 remain largely unknown. In this study, we generated GPAT2-deficient mice and conducted comprehensive analyses. The deficient mice showed defective piRNA production and subsequent de-silencing of IAP and Line-1 retrotransposons in fetal testes. In addition, apoptosis of pachytene spermatocytes was observed. These abnormalities were all common to the phenotype of PIWIL2-deficient mice, in which piRNA production was impaired. GPAT2-deficient mice exhibited apoptosis in spermatogonia at the neonatal stage, which was not observed in PIWIL2-deficient mice. These data show that GPAT2 plays a critical role in preventing apoptosis in spermatogonia.

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Female mice lacking Ftx lncRNA exhibit impaired X-chromosome inactivation and a microphthalmia-like phenotype

Hosoi Y., Soma M., Shiura H., Sado T., Hasuwa H., Abe K., Kohda T., Ishino F., Kobayashi S.

Nature Communications (Nature Communications)  9 ( 1 )  2018.12

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© 2018, The Author(s). X-chromosome inactivation (XCI) is an essential epigenetic process in female mammalian development. Although cell-based studies suggest the potential importance of the Ftx long non-protein-coding RNA (lncRNA) in XCI, its physiological roles in vivo remain unclear. Here we show that targeted deletion of X-linked mouse Ftx lncRNA causes eye abnormalities resembling human microphthalmia in a subset of females but rarely in males. This inheritance pattern cannot be explained by X-linked dominant or recessive inheritance, where males typically show a more severe phenotype than females. In Ftx-deficient mice, some X-linked genes remain active on the inactive X, suggesting that defects in random XCI in somatic cells cause a substantially female-specific phenotype. The expression level of Xist, a master regulator of XCI, is diminished in females homozygous or heterozygous for Ftx deficiency. We propose that loss-of-Ftx lncRNA abolishes gene silencing on the inactive X chromosome, leading to a female microphthalmia-like phenotype.

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Vwf K1362A resulted in failure of protein synthesis in mice

Sanda N., Suzuki N., Suzuki A., Kanematsu T., Kishimoto M., Hasuwa H., Takagi A., Kojima T., Matsushita T., Nakamura S.

International Journal of Hematology (International Journal of Hematology)  107 ( 4 ) 428 - 435 2018.04

ISSN  09255710

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© 2018, The Japanese Society of Hematology. Von Willebrand factor (VWF) is synthesized in megakaryocytes and endothelial cells (ECs) and has two main roles: to carry and protect coagulation factor VIII (FVIII) from degradation by forming VWF–FVIII complex; and to mediate platelet adhesion and aggregation at sites of vascular injury. Previous research using the HEK293 cell line revealed that the VWF K1362 mutation interacted directly with platelet glycoprotein Ib (GPIb). Vwf K1362A knock-in (KI) mice were therefore generated to verify the in vivo function of residue 1362 in binding to platelet GPIb. The Cre-loxP system was employed to introduce the Vwf K1362A mutation systemically in mice. In blood coagulation analysis, the VWF antigen (VWF:Ag) of Lys1362Ala KI homozygous (homo) mice was below the sensitivity of detection by enzyme-linked immunosorbent assay. FVIII activities (FVIII:C) were 47.9 ± 0.3 and 3.3 ± 0.3% (K1362A heterozygous (hetero) and K1362A KI homo mice, respectively) compared to wild-type mice. Immunohistochemical staining analysis revealed that VWF protein did not exist in ECs of K1362A KI homo mice. These results indicated that VWF protein synthesis of K1362A was impaired after transcription in mice. K1362 seems to represent a very important position not only for VWF function, but also for VWF synthesis in mice.

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Human PIWI (HIWI) is an azoospermia factor

Hasuwa H., Ishino K., Siomi H.

Science China Life Sciences (Science China Life Sciences)  61 ( 3 ) 348 - 350 2018.03

ISSN  16747305

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Constitutive overexpression of periostin delays wound healing in mouse skin

Nunomura S., Nanri Y., Ogawa M., Arima K., Mitamura Y., Yoshihara T., Hasuwa H., Conway S., Izuhara K.

Wound Repair and Regeneration (Wound Repair and Regeneration)  26 ( 1 ) 6 - 15 2018.01

ISSN  10671927

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© 2018 by the Wound Healing Society Periostin is a matricellular protein involved in development, maintenance, and regulation of tissues and organs via by binding to cell surface integrin receptors. Pathologically, periostin plays an important role in the process of wound healing: as a deficiency of the Postn gene delays wound closure and periostin is consistently up-regulated in response to injury and skin diseases. However, the functional role of elevated periostin in the process of wound healing has not been tested. In this study, we generated Postn-transgenic mice under the control of the CAG promoter/enhancer to investigate the effects of constitutive overexpression of full length periostin during its pathophysiological roles. Transgenic mice showed significant overexpression of periostin in skin, lung, and heart, but no morphological changes were observed. However, when these transgenic mice were injured, periostin overexpression delayed the closure of excisional wounds. Expression of IL-1β and TNFα, pro-inflammatory cytokines important for wound healing, was significantly decreased in the transgenic mice, prior to delayed healing. Infiltration of neutrophils and macrophages, the main sources of IL-1β and TNFα, was also down-regulated in the transgenic wound sites. From these data, we conclude that enforced expression of periostin delays wound closure due to reduced infiltration of neutrophils and macrophages followed by down-regulation of IL-1β and TNFα expression. This suggests that regulated spatiotemporal expression of periostin is important for efficient wound healing and that constitutive periostin overexpression interrupts the normal process of wound closure.

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Laminin γ1 c-terminal Glu to Gln mutation induces early postimplantation lethality

Kiyozumi D., Taniguchi Y., Nakano I., Toga J., Yagi E., Hasuwa H., Ikawa M., Sekiguchi K.

Life Science Alliance (Life Science Alliance)  1 ( 5 )  2018

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© 2018 Kiyozumi et al. Laminin–integrin interactions regulate various adhesion-dependent cellular processes. γ1C-Glu, the Glu residue in the laminin γ1 chain C-terminal tail, is crucial for the binding of γ1-laminins to several integrin isoforms. Here, we investigated the impact of γ1C Glu to Gln mutation on γ1-laminin binding to all possible integrin partners in vitro, and found that the mutation specifically ablated binding to α3, α6, and α7 integrins. To examine the physiological significance of γ1C-Glu, we generated a knock-in allele, Lamc1EQ, in which the γ1C Glu to Gln mutation was introduced. Although Lamc1EQ/EQ homozygotes developed into blastocysts and deposited laminins in their basement membranes, they died just after implantation because of disordered extraembryonic development. Given the impact of the Lamc1EQ allele on embryonic development, we developed a knock-in mouse strain enabling on-demand introduction of the γ1C Glu to Gln mutation by the Cre-loxP system. The present study has revealed a crucial role of γ1C-Glu–mediated integrin binding in postimplantation development and provides useful animal models for investigating the physiological roles of laminin–integrin interactions in vivo.

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Induction of DNA Methylation by Artificial piRNA Production in Male Germ Cells

Itou, Daisuke, Shiromoto, Yusuke, Shin-ya, Yukiho, Ishii, Chika, Nishimura, Toru, Ogonuki, Narumi, Ogura, Atsuo, Hasuwa, Hidetoshi, Fujihara, Yoshitaka, Kuramochi-Miyagawa, Satomi, Nakano, Toru

CURRENT BIOLOGY 25 ( 7 ) 901 - 906 2015.03

Research paper (scientific journal),  ISSN  0960-9822

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IN VIVO VISUALIZATION OF SUBTLE, TRANSIENT AND LOCAL ACTIVITY OF ASTROCYTES WITH ULTRASENSITIVE CALCIUM INDICATOR, YC-NANO50

Kanemaru, K., Sekiya, H., Xu, M., Satoh, K., Okubo, Y., Sasaki, T., Moritoh, S., Hasuwa, H., Mimura, M., Horikawa, K., Matsui, K., Nagai, T., Tanaka, K., Iino, M.

BASIC & CLINICAL PHARMACOLOGY & TOXICOLOGY 115   197 - 197 2014.07

Research paper (scientific journal),  ISSN  1742-7835

Optogenetic Manipulation of Activity and Temporally Controlled Cell-Specific Ablation Reveal a Role for MCH Neurons in Sleep/Wake Regulation

Tsunematsu, Tomomi, Ueno, Takafumi, Tabuchi, Sawako, Inutsuka, Ayumu, Tanaka, Kenji F., Hasuwa, Hidetoshi, Kilduff, Thomas S., Terao, Akira, Yamanaka, Akihiro

JOURNAL OF NEUROSCIENCE 34 ( 20 ) 6896 - 6909 2014.05

Research paper (scientific journal),  ISSN  0270-6474

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Seminal vesicle protein SVS2 is required for sperm survival in the uterus

Kawano, Natsuko, Araki, Naoya, Yoshida, Kaoru, Hibino, Taku, Ohnami, Naoko, Makino, Maako, Kanai, Seiya, Hasuwa, Hidetoshi, Yoshida, Manabu, Miyado, Kenji, Umezawa, Akihiro

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 111 ( 11 ) 4145 - 4150 2014.03

Research paper (scientific journal),  ISSN  0027-8424

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Interleukin-16 Promotes Cardiac Fibrosis and Myocardial Stiffening in Heart Failure with Preserved Ejection Fraction

Tamaki, Shunsuke, Mano, Toshiaki, Sakata, Yasushi, Ohtani, Tomohito, Takeda, Yasuharu, Kamimura, Daisuke, Omori, Yosuke, Tsukamoto, Yasumasa, Ikeya, Yukitoshi, Kawai, Mari, Kumanogoh, Atsushi, Hagihara, Keisuke, Ishii, Ryohei, Higashimori, Mitsuru, Kaneko, Makoto, Hasuwa, Hidetoshi, Miwa, Takeshi, Yamamoto, Kazuhiro, Komuro, Issei

PLOS ONE 8 ( 7 ) 71 - 73 2013.07

Research paper (scientific journal),  ISSN  1932-6203

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MiR-200b and miR-429 Function in Mouse Ovulation and Are Essential for Female Fertility

Hasuwa, Hidetoshi, Ueda, Jun, Ikawa, Masahito, Okabe, Masaru

SCIENCE 341 ( 6141 ) 71 - 73 2013.07

Research paper (scientific journal),  ISSN  0036-8075

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Expanding the Repertoire of Optogenetically Targeted Cells with an Enhanced Gene Expression System

Tanaka, Kenji F., Matsui, Ko, Sasaki, Takuya, Sano, Hiromi, Sugio, Shouta, Fan, Kai, Hen, Rene, Nakai, Junichi, Yanagawa, Yuchio, Hasuwa, Hidetoshi, Okabe, Masaru, Deisseroth, Karl, Ikenaka, Kazuhiro, Yamanaka, Akihiro

CELL REPORTS 2 ( 2 ) 397 - 406 2012.08

Research paper (scientific journal),  ISSN  2211-1247

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Enhanced response of T lymphocytes from Pgap3 knockout. mouse: Insight into roles of fatty acid remodeling of GPI anchored proteins

Murakami, Hidekazu, Wang, Yetao, Hasuwa, Hidetoshi, Maeda, Yusuke, Kinoshita, Taroh, Murakami, Yoshiko

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 417 ( 4 ) 1235 - 1241 2012.01

Research paper (scientific journal),  ISSN  0006-291X

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Transgenic Mouse Sperm that Have Green Acrosome and Red Mitochondria Allow Visualization of Sperm and Their Acrosome Reaction in Vivo

Hasuwa, Hidetoshi, Muro, Yuko, Ikawa, Masahito, Kato, Noriko, Tsujimoto, Yoshihide, Okabe, Masaru

EXPERIMENTAL ANIMALS 59 ( 1 ) 105 - 107 2010.01

Research paper (scientific journal),  ISSN  1341-1357

Targeted Gene Modification in Mouse ES Cells Using Integrase-Defective Lentiviral Vectors

Okada, Yuka, Ueshin, Yuko, Hasuwa, Hidetoshi, Takumi, Kazuhiro, Okabe, Masaru, Ikawa, Masahito

GENESIS 47 ( 4 ) 217 - 223 2009.04

Research paper (scientific journal),  ISSN  1526-954X

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Cd52, known as a major maturation-associated sperm membrane antigen secreted from the epididymis, is not required for fertilization in the mouse

Yamaguchi, Ryo, Yamagata, Kazuo, Hasuwa, Hidetoshi, Inano, Emiko, Ikawa, Masahito, Okabe, Masaru

GENES TO CELLS 13 ( 8 ) 851 - 861 2008.08

Research paper (scientific journal),  ISSN  1356-9597

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Heparin-binding EGF-like growth factor is a promising target for ovarian cancer therapy

Miyamoto, S, Hirata, M, Yamazaki, A, Kageyama, T, Hasuwa, H, Mizushima, H, Tanaka, Y, Yagi, H, Sonoda, K, Kai, M, Kanoh, H, Nakano, H, Mekada, E

CANCER RESEARCH 64 ( 16 ) 5720 - 5727 2004.08

Research paper (scientific journal),  ISSN  0008-5472

Small interfering RNA and gene silencing in transgenic mice and rats

Hasuwa, H, Kaseda, K, Einarsdottir, T, Okabe, M

FEBS LETTERS 532 ( 1-2 ) 227 - 230 2002.12

Research paper (scientific journal),  ISSN  0014-5793

Identification of mammalian TOM22 as a subunit of the preprotein translocase of the mitochondrial outer membrane

Saeki, K, Suzuki, H, Tsuneoka, M, Maeda, M, Iwamoto, R, Hasuwa, H, Shida, S, Takahashi, T, Sakaguchi, M, Endo, T, Miura, Y, Mekada, E, Mihara, K

JOURNAL OF BIOLOGICAL CHEMISTRY 275 ( 41 ) 31996 - 32002 2000.10

Research paper (scientific journal),  ISSN  0021-9258