Updated on 2025/09/30

写真a

 
WAKAI Takuya
 
Organization
Faculty of Environmental, Life, Natural Science and Technology Associate Professor
Position
Associate Professor
External link

Degree

  • 博士（農学） ( 東北大学 )

Research Interests

  • Mitochondria

  • Oocyte

  • Calcium signaling

  • Embryo development

  • Fertilization

  • Organelle

  • Endoplasmic Reticulum

Research Areas

  • Life Science / Animal production science

  • Life Science / Obstetrics and gynecology

  • Life Science / Developmental biology

  • Life Science / Applied molecular and cellular biology

  • Life Science / Cell biology

Research History

  • Okayama University   環境生命科学研究科   Associate Professor

    2014.12

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  • Tokyo University of Agriculture   Faculty of Applied Bio-Science

    2011.4 - 2014.11

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  • マサチューセッツ州立大学アムハースト校   博士研究員

    2007.7 - 2011.3

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Professional Memberships

  • THE MOLECULAR BIOLOGY SOCIETY OF JAPAN

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  • JAPAN SOCIETY FOR OVA RESEARCH

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  • SOCIETY FOR REPRODUCTION AND DEVELOPMENT

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Papers

  • Exogenous expression of PGC-1α during in vitro maturation impairs the developmental competence of porcine oocytes

    Son Quang Do, Hai Thanh Nguyen, Takuya Wakai, Hiroaki Funahashi

    Theriogenology   228   30 - 36   2024.10

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    Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

    DOI： 10.1016/j.theriogenology.2024.07.021

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  • The impact of cumulus cell viability and pre-culture with the healthy cell mass on brilliant cresyl blue (BCB) staining assessment and meiotic competence of suboptimal porcine oocytes

    Wanniarachchige Tharindu Lakshitha Fonseka, Son Quang Do, Phong Ngoc Van, Hai Thanh Nguyen, Takuya Wakai, Hiroaki Funahashi

    Theriogenology   226   158 - 166   2024.9

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    Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

    DOI： 10.1016/j.theriogenology.2024.06.012

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  • Hippo pathway inactivation through subcellular localization of NF2/merlin in outer cells of mouse embryos. International journal

    Nanami Goda, Yui Ito, Shun Saito, Miyabi Suzuki, Hanako Bai, Masashi Takahashi, Takuya Wakai, Manabu Kawahara

    Development (Cambridge, England)   151 ( 14 )   2024.7

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    Language：English   Publishing type：Research paper (scientific journal)  

    The Hippo pathway plays a crucial role in cell proliferation and differentiation during tumorigenesis, tissue homeostasis and early embryogenesis. Scaffold proteins from the ezrin-radixin-moesin (ERM) family, including neurofibromin 2 (NF2; Merlin), regulate the Hippo pathway through cell polarity. However, the mechanisms underlying Hippo pathway regulation via cell polarity in establishing outer cells remain unclear. In this study, we generated artificial Nf2 mutants in the N-terminal FERM domain (L64P) and examined Hippo pathway activity by assessing the subcellular localization of YAP1 in early embryos expressing these mutant mRNAs. The L64P-Nf2 mutant inhibited NF2 localization around the cell membrane, resulting in YAP1 cytoplasmic translocation in the polar cells. L64P-Nf2 expression also disrupted the apical centralization of both large tumor suppressor 2 (LATS2) and ezrin in the polar cells. Furthermore, Lats2 mutants in the FERM binding domain (L83K) inhibited YAP1 nuclear translocation. These findings demonstrate that NF2 subcellular localization mediates cell polarity establishment involving ezrin centralization. This study provides previously unreported insights into how the orchestration of the cell-surface components, including NF2, LATS2 and ezrin, modulates the Hippo pathway during cell polarization.

    DOI： 10.1242/dev.202639

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  • Redistribution of fragmented mitochondria ensure symmetric organelle partitioning and faithful chromosome segregation in mitotic mouse zygotes

    Haruna Gekko, Ruri Nomura, Daiki Kuzuhara, Masato Kaneyasu, Genpei Koseki, Deepak Adhikari, Yasuyuki Mio, John Carroll, Tomohiro Kono, Hiroaki Funahashi, Takuya Wakai

    2024.6

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    Publisher：Cold Spring Harbor Laboratory  

    Abstract

    In cleavage-stage embryos, preexisting organelles partition evenly into daughter blastomeres without significant cell growth after symmetric cell division. The presence of mitochondrial DNA within mitochondria and its restricted replication during preimplantation development makes their inheritance particularly important. While chromosomes are precisely segregated by the mitotic spindle, the mechanisms controlling mitochondrial partitioning remain poorly understood. In this study, we investigate the mechanism by which Dynamin-related protein 1 (Drp1) controls the mitochondrial redistribution and partitioning during embryonic cleavage. Deletion of Drp1 in mouse zygotes causes marked mitochondrial aggregation, and the majority of embryos arrest at the 2-cell stage. Clumped mitochondria are located in the center of mitotic Drp1-depleted zygotes with less uniform distribution, thereby preventing their symmetric partitioning. Asymmetric mitochondrial inheritance is accompanied by functionally inequivalent blastomeres with biased ATP and endoplasmic reticulum Ca2+levels. We also find that marked mitochondrial centration in Drp1-depleted zygotes prevents the assembly of parental chromosomes, resulting in chromosome segregation defects and binucleation. Thus, mitochondrial fragmentation mediated by Drp1 ensure proper organelle positioning and partitioning into functional daughters during the first embryonic cleavage.

    Impact statement

    Depletion of Dynamin-related protein 1, a key regulator of mitochondrial fission, in mouse zygotes impair symmetric organelle partitioning and chromosome segregation leading to early developmental arrest.

    DOI： 10.1101/2024.06.13.598818

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  • Negative correlations of mitochondrial DNA copy number in commercial frozen bull spermatozoa with the motility parameters after thawing

    Hai Thanh Nguyen, Son Quang Do, Hiroshi Kobayashi, Takuya Wakai, Hiroaki Funahashi

    Theriogenology   210   154 - 161   2023.10

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    Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

    DOI： 10.1016/j.theriogenology.2023.07.027

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  • Photo-dependent cytosolic delivery of shRNA into a single blastomere in a mouse embryo. International journal

    Yuka Ikawa, Takuya Wakai, Hiroaki Funahashi, Tet Htut Soe, Kazunori Watanabe, Takashi Ohtsuki

    Scientific reports   13 ( 1 )   13050 - 13050   2023.8

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    Language：English   Publishing type：Research paper (scientific journal)  

    Single-cell-specific delivery of small RNAs, such as short hairpin RNA (shRNA) and small noncoding RNAs, allows us to elucidate the roles of specific upregulation of RNA expression and RNAi-mediated gene suppression in early embryo development. The photoinduced cytosolic dispersion of RNA (PCDR) method that we previously reported can introduce small RNAs into the cytosol of photoirradiated cells and enable RNA delivery into a single-cell in a spatiotemporally specific manner. However, the PCDR method has only been applied to planer cultured cells and not to embryos. This study demonstrated that the PCDR method can be utilized for photo-dependent cytosolic shRNA delivery into a single blastomere and for single blastomere-specific RNA interference in mouse embryos. Our results indicate that PCDR is a promising approach for studying the developmental process of early embryogenesis.

    DOI： 10.1038/s41598-023-40361-9

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  • Rapid thawing of frozen bull spermatozoa by transient exposure to 70 °C improves the viability, motility and mitochondrial health

    Hai Thanh Nguyen, Son Quang Do, Rukmali Athurupana, Takuya Wakai, Hiroaki Funahashi

    Animal Reproduction   20 ( 3 )   2023

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    Publishing type：Research paper (scientific journal)   Publisher：FapUNIFESP (SciELO)  

    DOI： 10.1590/1984-3143-ar2022-0127

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  • Cell-cycle dependent GATA2 subcellular localization in mouse 2-cell embryos

    Masaya Komatsu, Hayato Tsukahara, Hanako Bai, Masashi Takahashi, Takuya Wakai, Manabu Kawahara

    Biochemical and Biophysical Research Communications   584   1 - 6   2021.11

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    Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

    DOI： 10.1016/j.bbrc.2021.10.077

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  • The autophagic inducer and inhibitor display different activities on the meiotic and developmental competencies of porcine oocytes derived from small and medium follicles.

    Chiyuki Kohata-Ono, Takuya Wakai, Hiroaki Funahashi

    The Journal of reproduction and development   65 ( 6 )   527 - 532   2019.12

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    Language：English   Publishing type：Research paper (scientific journal)  

    This study aimed to examine the effect of rapamycin (autophagy inducer) and 3-methyladenine (3-MA, autophagy inhibitor) on the meiotic and developmental competencies of porcine oocytes derived from medium follicles (MF, 3-6 mm in diameter) and small follicles (SF, 1-2 mm in diameter) during in vitro maturation (IVM) process. The presence of 1 nM but not 10 nM rapamycin significantly increased the maturation rate of MF-derived oocytes (P < 0.05). However, the maturation rate of SF-derived oocytes was not affected by rapamycin at both concentrations (1 nM and 10 nM). The maturation rate of MF-derived oocytes decreased significantly (P < 0.05) in the presence of 0.2 mM but not 2 mM 3-MA than non-supplemented control. In contrast, in SF-derived oocytes, 3-MA at both 0.2 and 2 mM concentrations did not affect the maturation rates. The presence of 1 nM rapamycin significantly increased the blastocyst formation rate of MF-derived mature oocytes following parthenogenetic activation (P < 0.05). However, the blastocyst formation rate of SF-derived mature oocytes was not affected by the presence of rapamycin. The presence of 3-MA significantly reduced the blastocyst formation rate of MF-derived mature oocytes but did not change that of SF-derived oocytes. In conclusion, our study results show differences in activity of the autophagy inducer and inhibitor on the meiotic and developmental competencies of MF- and SF-derived porcine oocytes.

    DOI： 10.1262/jrd.2019-112

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  • The autophagic inducer and inhibitor display different activities on the meiotic and developmental competencies of porcine oocytes derived from small and medium follicles

    Chiyuki Kohata-Ono, Takuya Wakai, Hiroaki Funahashi

    JOURNAL OF REPRODUCTION AND DEVELOPMENT   65 ( 6 )   527 - 532   2019.12

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  • マウス卵母細胞におけるTrim-Away法を用いたリン酸化タンパク質の分解

    月向 はるな, 野村 瑠莉, 舟橋 弘晃, 若井 拓哉

    The Journal of Reproduction and Development   65 ( Suppl. )   j119 - j119   2019.9

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    Language：Japanese   Publisher：(公社)日本繁殖生物学会  

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  • ミトコンドリア分裂因子Drp1の阻害による着床前マウス胚の発生停止

    野村 瑠莉, 月向 はるな, 舟橋 弘晃, 若井 拓哉

    The Journal of Reproduction and Development   65 ( Suppl. )   j120 - j120   2019.9

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    Language：Japanese   Publisher：(公社)日本繁殖生物学会  

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  • Ca2+ Signaling and Homeostasis in Mammalian Oocytes and Eggs. Reviewed

    Wakai T, Mehregan A, Fissore RA

    Cold Spring Harbor perspectives in biology   2019.8

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  • Removal of cumulus cells around 20 h after the start of in vitro maturation improves the meiotic competence of porcine oocytes via reduction in cAMP and cGMP levels. Reviewed

    Ferré-Pujol P, Nguyen XK, Nagahara T, Bui TTM, Wakai T, Funahashi H

    The Journal of reproduction and development   65 ( 2 )   177 - 182   2019.4

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    DOI： 10.1262/jrd.2018-130

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  • 着床前胚発生におけるミトコンドリア分裂因子Drp1の機能解析

    野村 瑠莉, 月向 はるな, 舟橋 弘晃, 若井 拓哉

    Journal of Mammalian Ova Research   36 ( 1 )   S43 - S43   2019.4

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    Language：Japanese   Publisher：(一社)日本卵子学会  

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  • Trim-Away法を用いたマウス卵母細胞におけるDrp1の機能解析

    月向 はるな, 野村 瑠莉, 舟橋 弘晃, 若井 拓哉

    Journal of Mammalian Ova Research   36 ( 1 )   S44 - S44   2019.4

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    Language：Japanese   Publisher：(一社)日本卵子学会  

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  • 体外成熟時のブタ卵母細胞のオートファジー能は由来する小中卵胞で異なる

    小野 千由貴, 若井 拓哉, 舟橋 弘晃

    Journal of Mammalian Ova Research   36 ( 1 )   S56 - S56   2019.4

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  • XY oocytes of sex-reversed females with a Sry mutation deviate from the normal developmental process beyond the mitotic stage†. Reviewed

    Sakashita A, Wakai T, Kawabata Y, Nishimura C, Sotomaru Y, Alavattam KG, Namekawa SH, Kono T

    Biology of reproduction   100 ( 3 )   697 - 710   2019.3

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    Publishing type：Research paper (scientific journal)   Publisher：Oxford University Press (OUP)  

    <title>Abstract</title>
    The fertility of sex-reversed XY female mice is severely impaired by a massive loss of oocytes and failure of meiotic progression. This phenomenon remains an outstanding mystery. We sought to determine the molecular etiology of XY oocyte dysfunction by generating sex-reversed females that bear genetic ablation of Sry, a vital sex determination gene, on an inbred C57BL/6 background. These mutant mice, termed XYsry− mutants, showed severe attrition of germ cells during fetal development, resulting in the depletion of ovarian germ cells prior to sexual maturation. Comprehensive transcriptome analyses of primordial germ cells (PGCs) and postnatal oocytes demonstrated that XYsry− females had deviated significantly from normal developmental processes during the stages of mitotic proliferation. The impaired proliferation of XYsry− PGCs was associated with aberrant β-catenin signaling and the excessive expression of transposable elements. Upon entry to the meiotic stage, XYsry− oocytes demonstrated extensive defects, including the impairment of crossover formation, the failure of primordial follicle maintenance, and no capacity for embryo development. Together, these results suggest potential molecular causes for germ cell disruption in sex-reversed female mice, thereby providing insights into disorders of sex differentiation in humans, such as “Swyer syndrome,” in which patients with an XY karyotype present as typical females and are infertile.

    DOI： 10.1093/biolre/ioy214

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  • Constitutive IP3R1-mediated Ca2+ release reduces Ca2+ store content and stimulates mitochondrial metabolism in mouse GV oocytes. Reviewed

    Wakai T, Fissore RA

    Journal of cell science   132 ( 3 )   2019.2

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  • Fertilization and the signaling of egg activation

    Takuya Wakai, Aujan Mehregan, Rafael A. Fissore

    Encyclopedia of Reproduction   368 - 375   2018.1

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    Language：English   Publishing type：Part of collection (book)   Publisher：Elsevier  

    DOI： 10.1016/B978-0-12-801238-3.64656-1

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  • Presence of vascular endothelial growth factor during the first half of IVM improves the meiotic and developmental competence of porcine oocytes from small follicles Reviewed

    Tra M. T. Bui, Khank X. Nguyen, Asako Karata, Pilar Ferre, Minh T. Tran, Takuya Wakai, Hiroaki Funahashi

    REPRODUCTION FERTILITY AND DEVELOPMENT   29 ( 10 )   1902 - 1909   2017.9

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    DOI： 10.1071/RD16321

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  • Levels of cyclic-AMP and cyclic-GMP in porcine oocyte-cumulus complexes and cumulus-free oocytes derived from small and middle follicles during the first 24-hour period of in vitro maturation Reviewed

    Yuichi Okudaira, Takuya Wakai, Hiroaki Funahashi

    JOURNAL OF REPRODUCTION AND DEVELOPMENT   63 ( 2 )   191 - 197   2017.4

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    DOI： 10.1262/jrd.2016-156

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  • Effect of removing cumulus cells from porcine cumulus-oocyte complexes derived from small and medium follicles during IVM on the apoptotic status and meiotic progression of the oocytes Reviewed

    Pilar Ferre, Tra Mi Thi Bui, Takuya Wakai, Hiroaki Funahashi

    THERIOGENOLOGY   86 ( 7 )   1705 - 1710   2016.10

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    DOI： 10.1016/j.theriogenology.2016.05.024

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  • DNA Methylation Errors in Cloned Mouse Sperm by Germ Line Barrier Evasion Reviewed

    Tasuku Koike, Takuya Wakai, Yuko Jincho, Akihiko Sakashita, Hisato Kobayashi, Eiji Mizutani, Sayaka Wakayama, Fumihito Miura, Takashi Ito, Tomohiro Kono

    BIOLOGY OF REPRODUCTION   94 ( 6 )   128   2016.6

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    DOI： 10.1095/biolreprod.116.138677

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  • A phosphodiesterase type-5 inhibitor, sildenafil, induces sperm capacitation and penetration into porcine oocytes in a chemically defined medium Reviewed

    Sumire Ioki, Qing-Shan Wu, Osamu Takayama, Hideyuki H. Motohashi, Takuya Wakai, Hiroaki Funahashi

    THERIOGENOLOGY   85 ( 3 )   428 - 433   2016.2

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    DOI： 10.1016/j.theriogenology.2015.09.013

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  • Detection of DNA Methylation Errors in Sperm of Somatic Cell Cloned Mice

    KOIKE Tasuku, KONO Tomohiro, JINCHO Yuko, WAKAI Takuya, MIZUTANI Eiji, KOBAYASHI Hisato, WAKAYAMA Sayaka, SAKASHITA Akihiko, MIURA Fumihito, ITO Takashi

    The Journal of Reproduction and Development Supplement   108 ( 0 )   P - 82-P-82   2015

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    Language：Japanese   Publisher：THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT  

    DOI： 10.14882/jrds.108.0_P-82

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  • Mitochondrial dynamics controlled by mitofusins define organelle positioning and movement during mouse oocyte maturation Reviewed

    Takuya Wakai, Yuichirou Harada, Kenji Miyado, Tomohiro Kono

    MOLECULAR HUMAN REPRODUCTION   20 ( 11 )   1090 - 1100   2014.11

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    DOI： 10.1093/molehr/gau064

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  • Dynamics of genomic 5-hydroxymethylcytosine during mouse oocyte growth Reviewed

    Akihiko Sakashita, Hisato Kobayashi, Takuya Wakai, Yusuke Sotomaru, Kenichiro Hata, Tomohiro Kono

    GENES TO CELLS   19 ( 8 )   629 - 636   2014.8

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    DOI： 10.1111/gtc.12164

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  • Forced expression of DNA methyltransferases during oocyte growth accelerates the establishment of methylation imprints but not functional genomic imprinting Reviewed

    Satoshi Hara, Takashi Takano, Tsugunari Fujikawa, Munehiro Yamada, Takuya Wakai, Tomohiro Kono, Yayoi Obata

    HUMAN MOLECULAR GENETICS   23 ( 14 )   3853 - 3864   2014.7

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    DOI： 10.1093/hmg/ddu100

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  • Long exposure to mature ooplasm can alter DNA methylation at imprinted loci in non-growing oocytes but not in prospermatogonia （成熟卵細胞質に非成長期卵母細胞核を暴露するとインプリント領域のメチル化に変化が生じるが精原細胞の核を暴露した場合では変化が生じない） Reviewed

    Obata Y, Wakai T, Hara S, Kono T

    Reproduction   147 ( 1 )   H1 - H6   2014.1

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    精子あるいは卵子特異的インプリント確立機構を解明する為に、卵母細胞のインプリントが確立する条件下に雄性生殖細胞を暴露した。十分成長した卵母細胞に雄性生殖細胞あるいは非成長期卵母細胞を核移植し、再構築卵胞を形成させ体外培養を実施した。その結果、精子型インプリントを確立した後の雄性生殖細胞には、もはや卵子型インプリントを書き込むことはできなかった。一方、卵子型インプリントが確立する前の非成長期卵母細胞を移植した場合、この卵母細胞ゲノムには精子型インプリントも卵子型インプリントも書き込まれることが示された。このことから、卵子特異的なインプリントは、精子特異的なインプリントが書き込まれないよう、ブロックする機構であることが示唆された。

    DOI： 10.1530/REP-13-0359.

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  • Long exposure to mature ooplasm can alter DNA methylation at imprinted loci in non-growing oocytes but not in prospermatogonia Reviewed

    Yayoi Obata, Takuya Wakai, Satoshi Hara, Tomohiro Kono

    REPRODUCTION   147 ( 1 )   H1 - H6   2014.1

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    DOI： 10.1530/REP-13-0359

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  • Regulation of endoplasmic reticulum Ca2+ oscillations in mammalian eggs Reviewed

    Takuya Wakai, Nan Zhang, Peter Vangheluwe, Rafael A. Fissore

    JOURNAL OF CELL SCIENCE   126 ( 24 )   5714 - 5724   2013.12

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    DOI： 10.1242/jcs.136549

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  • Artificial Activation of Mammalian Oocytes for Cloning: Present Status and Future Perspectives Reviewed

    Takuya Wakai, Junya Ito, Rafael A. Fissore

    Principles of Cloning: Second Edition   3 - 10   2013.10

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    Language：English   Publishing type：Part of collection (book)   Publisher：Elsevier Inc.  

    DOI： 10.1016/B978-0-12-386541-0.00001-1

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  • Ca2+ influx and the store-operated Ca2+ entry pathway undergo regulation during mouse oocyte maturation Reviewed

    Banyoon Cheon, Hoi-Chang Lee, Takuya Wakai, Rafael A. Fissore

    MOLECULAR BIOLOGY OF THE CELL   24 ( 9 )   1396 - 1410   2013.5

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    DOI： 10.1091/mbc.E13-01-0065

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  • High-resolution DNA methylome analysis of primordial germ cells identifies gender-specific reprogramming in mice Reviewed

    Hisato Kobayashi, Takayuki Sakurai, Fumihito Miura, Misaki Imai, Kentaro Mochiduki, Eikichi Yanagisawa, Akihiko Sakashita, Takuya Wakai, Yutaka Suzuki, Takashi Ito, Yasuhisa Matsui, Tomohiro Kono

    GENOME RESEARCH   23 ( 4 )   616 - 627   2013.4

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    DOI： 10.1101/gr.148023.112

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  • Ca2+ homeostasis and regulation of ER Ca2+ in mammalian oocytes/eggs Reviewed

    Takuya Wakai, Rafael A. Fissore

    CELL CALCIUM   53 ( 1 )   63 - 67   2013.1

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  • Molecular characteristics of horse phospholipase C zeta (PLC) Reviewed

    Kana Sato, Takuya Wakai, Yasunari Seita, Akiko Takizawa, Rafael A. Fissore, Junya Ito, Naomi Kashiwazaki

    ANIMAL SCIENCE JOURNAL   84 ( 4 )   359 - 368   2013

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    DOI： 10.1111/asj.12044

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  • Regulation of inositol 1,4,5-trisphosphate receptor function during mouse oocyte maturation Reviewed

    Takuya Wakai, Veerle Vanderheyden, Sook-Young Yoon, Banyoon Cheon, Nan Zhang, Jan B. Parys, Rafael A. Fissore

    JOURNAL OF CELLULAR PHYSIOLOGY   227 ( 2 )   705 - 717   2012.2

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    DOI： 10.1002/jcp.22778

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  • Caffeine Alleviates the Deterioration of Ca2+ Release Mechanisms and Fragmentation of In Vitro-Aged Mouse Eggs Reviewed

    Nan Zhang, Takuya Wakai, Rafael A. Fissore

    MOLECULAR REPRODUCTION AND DEVELOPMENT   78 ( 9 )   684 - 701   2011.9

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    DOI： 10.1002/mrd.21366

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  • Ca2+ Signaling During Mammalian Fertilization: Requirements, Players, and Adaptations Reviewed

    Takuya Wakai, Veerle Vanderheyden, Rafael A. Fissore

    COLD SPRING HARBOR PERSPECTIVES IN BIOLOGY   3 ( 4 )   2011.4

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    DOI： 10.1101/cshperspect.a006767

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  • Anomalous Oxygen Consumption in Porcine Somatic Cell Nuclear Transfer Embryos Reviewed

    Satoshi Sugimura, Masaki Yokoo, Ken-ichi Yamanaka, Manabu Kawahara, Satoru Moriyasu, Takuya Wakai, Takashi Nagai, Hiroyuki Abe, Eimei Sato

    CELLULAR REPROGRAMMING   12 ( 4 )   463 - 474   2010.8

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    DOI： 10.1089/cell.2009.0111

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  • Early metaphase II oocytes treated with dibutyryl cyclic adenosine monophosphate provide suitable recipient cytoplasm for the production of miniature pig somatic cell nuclear transfer embryos Reviewed

    Satoshi Sugimura, Ken-ichi Yamanaka, Manabu Kawahara, Takuya Wakai, Masaki Yokoo, Eimei Sato

    ANIMAL SCIENCE JOURNAL   81 ( 1 )   48 - 57   2010

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    DOI： 10.1111/j.1740-0929.2009.00705.x

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  • Phosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes Reviewed

    Junya Ito, Tomoko Yoshida, Yasushi Kasai, Takuya Wakai, Jan B. Parys, Rafael A. Fissore, Naomi Kashiwazaki

    ANIMAL SCIENCE JOURNAL   81 ( 1 )   34 - 41   2010

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    DOI： 10.1111/j.1740-0929.2009.00699.x

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  • Acetylation Level of Histone H3 in Early Embryonic Stages Affects Subsequent Development of Miniature Pig Somatic Cell Nuclear Transfer Embryos Reviewed

    Ken-ichi Yamanaka, Satoshi Sugimura, Takuya Wakai, Manabu Kawahara, Eimei Sato

    JOURNAL OF REPRODUCTION AND DEVELOPMENT   55 ( 6 )   638 - 644   2009.12

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    DOI： 10.1262/jrd.20245

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  • Profile of new green fluorescent protein transgenic Jinhua pigs as an imaging source Reviewed

    Tatsuo Kawarasaki, Kazuhiko Uchiyama, Atsushi Hirao, Sadahiro Azuma, Masayoshi Otake, Masatoshi Shibata, Seiko Tsuchiya, Shin Enosawa, Koichi Takeuchi, Kenjiro Konno, Yoji Hakamata, Hiroyuki Yoshino, Takuya Wakai, Shigeo Ookawara, Hozumi Tanaka, Eiji Kobayashi, Takashi Murakami

    JOURNAL OF BIOMEDICAL OPTICS   14 ( 5 )   054017   2009.9

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    DOI： 10.1117/1.3241985

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  • Regulation of inositol 1,4,5-trisphosphate receptor type 1 function during oocyte maturation by MPM-2 phosphorylation Reviewed

    Veerle Vanderheyden, Takuya Wakai, Geert Bultynck, Humbert De Smedt, Jan B. Parys, Rafael A. Fissore

    CELL CALCIUM   46 ( 1 )   56 - 64   2009.7

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    DOI： 10.1016/j.ceca.2009.04.004

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  • Difference in Sensitivity to Culture Condition Between In Vitro Fertilized and Somatic Cell Nuclear Transfer Embryos in Pigs Reviewed

    Ken-ichi Yamanaka, Satoshi Sugimura, Takuya Wakai, Manabu Kawahara, Eimei Sato

    JOURNAL OF REPRODUCTION AND DEVELOPMENT   55 ( 3 )   299 - 304   2009.6

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    DOI： 10.1262/jrd.20174

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  • Production of viable cloned miniature pig embryos using oocytes derived from domestic pig ovaries Reviewed

    Takuya Wakai, Satoshi Sugimura, Ken-Ichi Yamanaka, Manabu Kawahara, Hiroshi Sasada, Hozumi Tanaka, Asako Ando, Eiji Kobayashi, Eimei Sato

    CLONING AND STEM CELLS   10 ( 2 )   249 - 261   2008.6

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    DOI： 10.1089/clo.2007.0045

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  • Effect of cytochalasins B and D on the developmental competence of somatic cell nuclear transfer embryos in miniature pigs. Reviewed

    Sugimura S, Kawahara M, Wakai T, Yamanaka K, Sasada H, Sato E

    Zygote (Cambridge, England)   16 ( 2 )   153 - 159   2008.5

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  • Induction of estrus in pubertal miniature gilts Reviewed

    Takuya Wakai, Hozumi Tanaka, Ken-ichi Yamanaka, Satoshi Sugimura, Hiroshi Sasada, Manabu Kawahara, Eiji Kobayashi, Eimei Sato

    ANIMAL REPRODUCTION SCIENCE   103 ( 1-2 )   193 - 198   2008.1

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    DOI： 10.1016/j.anireprosci.2007.04.004

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  • Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos Reviewed

    Ken-ichi Yamanaka, Satoshi Sugimura, Takuya Wakai, Takehisa Shoji, Jin Kobayashi, Hiroshi Sasada, Eimei Sato

    JOURNAL OF REPRODUCTION AND DEVELOPMENT   53 ( 4 )   791 - 800   2007.8

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    DOI： 10.1262/jrd.18162

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  • Recent patents of TGF-beta family and VEGF associated with ovarian follicular development in mammals. Reviewed

    Shimizu T, Abe Y, Wakai T, Hoshino Y, Miyamoto A, Sato E

    Recent patents on DNA & gene sequences   1 ( 3 )   195 - 199   2007

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  • Recent patents of TGF-beta family and VEGF associated with ovarian follicular development in mammals. Reviewed

    Shimizu Takashi, Abe Yasuyuki, Wakai Takuya, Hoshino Yumi, Miyamoto Akio, Sato Eimei

    Recent patents on DNA & gene sequences   1 ( 3 )   195 - 199   2007

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  • Caffeine promotes premature chromosome condensation formation and in vitro development in porcine reconstructed embryos via a high level of maturation promoting factor activity during nuclear transfer Reviewed

    M Kawahara, T Wakai, KI Yamanaka, J Kobayashi, S Sugimura, T Shimizu, H Matsumoto, JH Kim, H Sasada, E Sato

    REPRODUCTION   130 ( 3 )   351 - 357   2005.9

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    DOI： 10.1530/rep.1.00644

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  • 哺乳動物初期胚への光による時空間特異的なRNA導入法

    大槻高史, 井川優風, 若井拓哉, 舟橋弘晃, 渡邉和則

    日本光医学・光生物学会   45th   2023

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  • Spatiotemporally photocontrolled RNA transfection in early mammalian embryos

    井川優風, 若井拓哉, 舟橋弘晃, 渡邉和則, 大槻高史

    日本バイオマテリアル学会大会予稿集(Web)   44th   2022

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  • Trim‐Away法を用いたマウス卵母細胞におけるDrp1の機能解析

    月向はるな, 野村瑠莉, 舟橋弘晃, 若井拓哉

    Journal of Mammalian Ova Research   36 ( 1 )   S44 - S44   2019.4

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  • 体外成熟時のブタ卵母細胞のオートファジー能は由来する小中卵胞で異なる

    小野千由貴, 若井拓哉, 舟橋弘晃

    Journal of Mammalian Ova Research   36 ( 1 )   S56 - S56   2019.4

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  • 着床前胚発生におけるミトコンドリア分裂因子Drp1の機能解析

    野村瑠莉, 月向はるな, 舟橋弘晃, 若井拓哉

    Journal of Mammalian Ova Research   36 ( 1 )   S43 - S43   2019.4

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  • ミトコンドリア分裂因子Drp1の阻害による着床前マウス胚の発生停止

    野村瑠莉, 月向はるな, 舟橋弘晃, 若井拓哉

    Journal of Reproduction and Development   65 ( Suppl Japanese Issue )   2019

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  • マウス卵母細胞におけるTrim-Away法を用いたリン酸化タンパク質の分解

    月向はるな, 野村瑠莉, 舟橋弘晃, 若井拓哉

    Journal of Reproduction and Development   65 ( Suppl Japanese Issue )   2019

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  • 光応答的に働く卵活性化因子の開発

    大本和正, 若井拓哉, 舟橋弘晃, 渡邉和則, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)   42nd   2019

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  • Dendra2蛍光タンパク質を用いた初期胚ミトコンドリアの細胞間分配の解析

    若井拓哉, 野村瑠璃

    Journal of Reproduction and Development   64 ( Suppl Japanese Issue )   j105 - j105   2018.9

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  • 初期胚におけるミトコンドリア分裂因子Drp1の役割

    野村瑠莉, 若井拓哉, 舟橋弘晃

    日本卵子学会誌   3 ( 1 )   S58 - S58   2018.4

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  • ブタ顕微授精卵での精子頭部膨化・雄性前核形成不全の解析

    林雄平, 若井拓哉, 舟橋弘晃

    日本卵子学会誌   3 ( 1 )   S45 - S45   2018.4

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  • 受精後のDNAメチル化リプログラミングにおける種間差異

    澤田友季乃, 舟橋弘晃, 若井拓哉

    岡山実験動物研究会報   ( 34 )   76‐77 - 77   2018.4

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  • Epidermal Growth Factorはヒト初期卵胞の体外発育を促進する

    高山修, 奥平裕一, 若井拓哉, 中塚幹也, 中塚幹也, 舟橋弘晃, 舟橋弘晃

    日本卵子学会誌   2 ( 1 )   S58 - S58   2017.4

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  • ウシ精巣からの精原幹細胞採取の試み

    永原知樹, 若井拓哉, 舟橋弘晃

    岡山実験動物研究会報   ( 33 )   50 - 50   2017.4

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  • ブタ卵母細胞中のミトコンドリア細胞内分布の変化

    葛原大貴, 若井拓哉, 舟橋弘晃

    岡山実験動物研究会報   ( 33 )   50‐51 - 51   2017.4

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  • 異なる還元剤による精子前処理がICSIブタ卵での雄性前核形成に及ぼす影響

    林雄平, 若井拓哉, 舟橋弘晃

    日本畜産学会大会講演要旨   122nd   156 - 156   2017.3

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  • Mitochondrial dynamics in mammalian oocytes

    若井拓哉

    岡山大学農学部学術報告(Web)   106   39‐42 (WEB ONLY)   2017.2

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  • Mitochondrial dynamics in mammalian oocytes

    若井 拓哉

    岡山大学農学部学術報告   106 ( 106 )   39 - 42   2017.2

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    Subcellular distribution of mitochondria are reorganized during development of the oocyte into afertilizable egg. In growing oocytes, mitochondria are heterogeneously distributed in the cytoplasm, whereas they diffuse throughout the cytoplasm in fully-grown oocytes. GFP-labeled mitochondria demonstrate that oocyte maturation involves dynamic redistribution of the mitochondria, whose process is associated with microtubule organization. These spatio-temporal regulations of mitochondria in oocytes may be an important process in the preparation for fertilization and subsequent embryonic development.

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  • 減数分裂と卵割におけるミトコンドリアの細胞間分配

    若井拓哉

    日本卵子学会誌   1 ( 1 )   S5   2016.4

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  • マウス卵子におけるミトコンドリア

    若井 拓哉

    岡山実験動物研究会報   ( 32 )   56 - 57   2016.4

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  • 精子・卵子研究から生殖補助医療への新たな展開 減数分裂と卵割におけるミトコンドリアの細胞間分配

    若井 拓哉

    日本卵子学会誌   1 ( 1 )   S5 - S5   2016.4

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  • マウス卵子におけるミトコンドリアの特性

    若井拓哉

    岡山実験動物研究会報   ( 32 )   9‐11 - 11   2016.4

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  • IVM中のブタ小中卵胞由来COC,裸化卵母細胞,卵丘細胞内のcAMPおよびcGMP量

    奥平裕一, 若井拓哉, 舟橋弘晃

    Journal of Reproduction and Development   61 ( Suppl Japanese Issue )   J91 - j91   2015.9

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  • ヒト小卵胞および中卵胞由来卵丘細胞‐卵母細胞複合体の体外成熟能

    奥平裕一, 高山修, BUI Thi Tra Mi, FERRE Pilar, 本橋秀之, 若井拓哉, 中塚幹也, 中塚幹也, 舟橋弘晃, 舟橋弘晃

    日本IVF学会雑誌   18 ( 2 )   81 - 81   2015.9

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  • Moesin遺伝子発現の可視化による体細胞クローン胚におけるX染色体リプログラミングの評価

    若井拓哉, 森園倫成, 渡辺大士, 外丸祐介, 神田暁史, 塩沢誠司, 河野友宏

    Journal of Reproduction and Development   60 ( Suppl Japanese Issue )   J142 - j142   2014.8

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  • 体細胞クローンマウス精子のDNAメチレーションエラー

    河野友宏, 若井拓哉, 水谷英二, 小林久人, 若山清香, 坂下陽彦, 三浦史仁, 伊藤隆司

    Journal of Reproduction and Development   60 ( Suppl Japanese Issue )   J75 - j75   2014.8

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    DOI： 10.14882/jrds.107.0_OR1-25

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  • マウス初期胚における5hmCおよびH3K9me2の挙動とDNA脱メチル化制御機構

    坂下陽彦, 中島芽生, 若井拓哉, 小林久人, 井関陽介, 河野友宏

    Journal of Mammalian Ova Research   31 ( 2 )   S78 - S78   2014.4

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  • 哺乳類卵子における小胞体Ca2+オシレーションの制御

    若井拓哉, 河野友宏, FISSORE Rafael

    Journal of Mammalian Ova Research   31 ( 2 )   S32   2014.4

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  • 体細胞クローンマウス精子のDNAメチル化リプログラミング

    若井拓哉, 水谷英二, 小林久人, 若山清香, 坂下陽彦, 伊藤隆司, 三浦史仁, 河野友宏

    日本分子生物学会年会プログラム・要旨集(Web)   37th   3W8-6(3P-0172) (WEB ONLY)   2014

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  • マウス生殖細胞・初期胚のDNAメチローム解析

    小林久人, 坂下陽彦, 若井拓哉, 小池佐, 佐野賢, 河野友宏

    日本分子生物学会年会プログラム・要旨集(Web)   37th   2W6-5(2P-0005) (WEB ONLY)   2014

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  • マウス卵母細胞におけるATP濃度の可視化と測定

    若井拓哉, 今村博臣, 河野友宏

    Journal of Reproduction and Development   59 ( Suppl Japanese Issue )   J140 - j140   2013.8

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    DOI： 10.14882/jrds.106.0.P-76.0

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  • マウス卵子形成過程における5ヒドロキシメチル化シトシンのダイナミクス

    坂下陽彦, 小林久人, 井関陽介, 若井拓哉, 外丸祐介, 河野友宏

    Journal of Reproduction and Development   59 ( Suppl Japanese Issue )   J77 - j77   2013.8

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    DOI： 10.14882/jrds.106.0.OR1-33.0

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  • マウス卵母細胞の成熟過程におけるミトコンドリア動態

    若井拓哉, 河野友宏

    Journal of Mammalian Ova Research   30 ( 2 )   S87 - S87   2013.4

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  • RNF12 is a Primary Factor for Xist Activation in Mouse Somatic Cloned Embryos

    FUKUDA Atsushi, WAKAI Takuya, WATANABE Hiroshi, KONO Tomohiro

    59 ( 1 )   A4   2013.2

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  • EIF1A expression in sertoli cell nuclear transfer embryos during preimplantation development

    Feng Cao, Takuya Wakai, Hiroshi Watanabe, Tomohiro Kono

    Journal of Mammalian Ova Research   30 ( 1 )   41 - 45   2013

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    DOI： 10.1274/jmor.30.41

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  • EIF1A expression in sertoli cell nuclear transfer embryos during preimplantation development

    Feng Cao, Takuya Wakai, Hiroshi Watanabe, Tomohiro Kono

    Journal of Mammalian Ova Research   30 ( 1 )   41 - 45   2013

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    DOI： 10.1274/jmor.30.41

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  • マウス卵子形成過程における5‐hydroxymethylcytosineのダイナミクス

    坂下陽彦, 小林久人, 井関陽介, 若井拓哉, 外丸祐介, 河野友宏

    日本分子生物学会年会プログラム・要旨集(Web)   36th   2P-0679 (WEB ONLY)   2013

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  • Mitochondrial Localization in Mouse Oocytes

    WAKAI Takuya

    Journal of mammalian ova research   29 ( 4 )   155 - 160   2012.10

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    卵母細胞が受精可能な卵子へと発育する過程でミトコンドリアの細胞内局在は大きく変動する。そこで,その局在の詳細を明らかにするために成長および成熟過程の卵母細胞におけるミトコンドリアの生細胞観察を行った。成長期の卵母細胞におけるミトコンドリアは細胞膜直下および核周辺に特に集中しているのに対して,成長を完了した卵母細胞では細胞質全体に拡散して分布しており,同一の組胞周期にある卵母細胞においても成長段階に応じてミトコンドリアの細胞内局在は異なることが分かった。また,成熟過程における卵母細胞のミトコンドリアをGFPでラベルし経時的に追跡したところ,局在のダイナミックな再構築が観察され,さらにそれは微小管の影響を受けていることが分かった。こうしたミトコンドリアの時間･空間的な制御は受精やその後の発生を支持するために必要と考えられる。

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  • 【卵子のエネルギー代謝-ミトコンドリア機能について-】マウス卵母細胞におけるミトコンドリアの細胞内局在

    若井 拓哉

    Journal of Mammalian Ova Research   29 ( 4 )   155 - 160   2012.10

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    Other Link： http://agriknowledge.affrc.go.jp/RN/2010834004

  • マウス始原生殖細胞におけるDNAメチローム解析

    小林久人, 櫻井隆順, 三浦史仁, 今井美咲, 望月研太郎, 柳澤永吉, 坂下陽彦, 若井拓哉, 鈴木穣, 伊藤隆司, 松居靖久, 河野友宏

    Journal of Reproduction and Development   58 ( Suppl Japanese Issue )   J102 - j102   2012.8

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  • Maternal RNF12 inhibits developmental reprogramming of somatic cell in mice

    FUKUDA Atsushi, WAKAI Takuya, WATANABE Hiroshi, KONO Tomohiro

    The Journal of Reproduction and Development Supplement   105 ( 0 )   j92 - j92   2012.8

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    DOI： 10.14882/jrds.105.0_5

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  • マウス卵子の減数分裂におけるH3K27me3の伝達機構

    若井拓哉, 河野友宏

    Journal of Mammalian Ova Research   29 ( 2 )   S55 - S55   2012.4

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  • マウス卵母細胞の成熟過程におけるミトコンドリアの動態

    若井拓哉, 河野友宏

    日本分子生物学会年会プログラム・要旨集(Web)   35th   4P-0399 (WEB ONLY)   2012

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  • Mitochondrial localization in mouse oocytes

    Takuya Wakai

    Journal of Mammalian Ova Research   29 ( 4 )   155 - 160   2012

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  • マウス始原生殖細胞におけるDNAメチローム解析

    小林久人, 櫻井隆順, 三浦史仁, 今井美咲, 望月研太郎, 柳澤永吉, 坂下陽彦, 若井拓哉, 鈴木穣, 伊藤隆司, 松居靖久, 河野友宏

    日本分子生物学会年会プログラム・要旨集(Web)   35th   1W1I-5 (WEB ONLY)   2012

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  • Mitochondrial Localization in Mouse Oocytes

    Wakai Takuya

    Journal of Mammalian Ova Research   29 ( 4 )   155 - 160   2012

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    Intracellular distribution of mitochondria varies during development of the oocyte into a fertilizable egg. Here we demonstrate a change in the distribution of mitochondria during oocyte growth and maturation in living mouse oocytes. In growing oocytes, mitochondria concentrated into subcortical and perinuclear areas, whereas it diffusely distributed throughout the cytoplasm in fully-grown oocytes, indicating the variation of mitochondrial distribution in the same G2/M cell-cycle stages. GFP-labeled mitochondria reveal that oocyte maturation involves dynamic reorganization of the mitochondria, and whose process is associated with microtubule organization. These spatio-temporal regulations of mitochondria in oocytes may be important process in the preparation of fertilization and subsequent embryonic development.&lt;br&gt;

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  • University of Massachusetts Amherst

    WAKAI Takuya, Fissore Rafael

    The Journal of Reproduction and Development Supplement   104 ( 0 )   215 - 215   2011.8

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  • Birth of somatic cell nuclear transfer(SCNT)-cloned miniature piglet following co-transfer parthenogenetic embryos

    Sugimura,S, Yamanaka,K, Wakai,T, Kawahara,M, Tanaka,H, Kobayashi,J, Kobayashi,E, Sato,E

    Tohoku J.Agr.Res.   59 ( 3,4 )   51 - 61   2009

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  • Vitrification of immature oocytes in domestic and endangered wild animals

    佐藤英明, 若井拓哉, 星野由美

    岩谷直治記念財団研究報告書   30   1 - 2   2007.8

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  • 初期胚発生におけるミトコンドリアの機能解析

    杉村智史, 横尾正樹, 山中賢一, 青野展也, 若井拓哉, 佐々田比呂志, 阿部宏之, 佐藤英明

    日本生殖医学会雑誌   52 ( 1/2 )   62 - 62   2007.4

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  • 医療目的の遺伝子改変家畜開発の到達点と今後

    若井拓哉, 佐藤英明

    畜産の研究   61 ( 4 )   437 - 441   2007.4

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  • ミニブタ体細胞クローン個体作出成功に関わる要因解析

    杉村 智史, 山中 賢一, 若井 拓哉, 佐藤 英明

    日本胚移植学雑誌   29 ( 1 )   10 - 13   2007.1

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  • Recent progress and perspectives of the research in developming transgenic animals

    Wakai,T, Sato,E

    Sustainable Livestock Production and Human Welfare   61 ( 4 )   437 - 441   2007

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  • Evaluation of mitochondrial function in single cloned miniature pig embryos by measuring oxygen consumption

    S. Sugimura, M. Yokoo, K. -I. Yamanaka, T. Wakai, H. Abe, T. Shoji, H. Sasada, J. Kobayashi, H. Abe, E. Sato

    REPRODUCTION FERTILITY AND DEVELOPMENT   19 ( 1 )   161 - 161   2007

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  • Recent advances in understanding the morphological changes in the meiotic maturation in mammalian cumulus-oocyte complexe

    Hoshino,Y, Yokoo,M, Wakai,T, Tomioka,I, Sato,E

    Morphology   2 ( 1 )   8 - 22   2007

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  • Morphological aspects of porcine embryos resonstructed by somatic cell nuclear transfer

    Wakai,T, Kawahara,M, Yamanaka,K, Hoshino,Y, Tomioka,I, Sugimura,S, Sugawara,A, Sato,E

    Morphology   2 ( 1 )   23 - 27   2007

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  • 単為発生卵との共移植による体細胞クローンミニブタの作出

    山中賢一, 杉村智史, 若井拓哉, 庄司孟央, 佐々田比呂志, 田中穂積, 小林英司, 佐藤英明

    日本畜産学会大会講演要旨   106th   86 - 86   2006.3

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  • ミニブタ体細胞クローン個体作出成功に関わる要因解析

    杉村智史, 山中賢一, 若井拓哉, 佐藤英明

    日本胚移植学雑誌   29 ( 1 )   10 - 13   2006.1

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  • Utility of parthenogenetic embryos as a co-transfer embryo for so nuclear transferred embryos in production of cloned miniature pigs

    Sugimura,S, Yamanaka,K, Wakai,T, Shouji,T, Sasada,H, Tanaka,H, Kobayashi,J, Kobayashi,E, Sato,E

    In Challenge of Animal Industry for the Wellbeing of Mankind, XIIth AAAP Animal Science Congress   1 - 3   2006

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  • Effect of culture media on in vitro development of SCNT-cloned miniature pig embryos

    Yamanaka,K, Sugimura,S, Wakai,T, Shoji,T, Kobayashi,J, Sasada,H, Sato,E

    In Challenge of Animal Industry for the Wellbeing of Mankind, XIIth AAAP Animal Science Congress 2006   9 - 11   2006

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  • ミャンマー連邦の畜産行政

    田中穂積, 若井拓哉, 金川弘司

    北海道獣医師会雑誌   49 ( 9 )   366 - 370   2005.9

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  • 自治医科大学におけるブタを用いた医学教育と研究

    田中穂積, 和久井亨, 若井拓哉, 吉野浩之, 河野正樹, 岡田真樹, 筒井真理子, 鳥取潤一, 小林英司

    日本獣医学会学術集会講演要旨集   140th   224 - 224   2005.8

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  • レシピエント卵子の体外成熟培養時間がミニブタ体細胞核の再構築およびクローンはいの発生に及ぼす影響

    杉村智史, 若井拓哉, 山中賢一, 庄司孟央, 佐々田比呂志, 佐藤英明

    Journal of Reproduction and Development   51 ( Supplement )   J104   2005.8

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  • アクチンフィラメントを指標としたミニブタ再構築はい活性化方法の最適化

    山中賢一, 杉村智史, 庄司孟央, 若井拓哉, 佐々田比呂志, 佐藤英明

    日本畜産学会大会講演要旨   105th   12   2005.8

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  • 低単位の性腺刺激ホルモンによるミニブタの発情誘起

    若井拓哉, 田中穂積, 山中賢一, 杉村智志, 筒井真理子, 鳥取潤一, 佐々田比呂志, 佐藤英明, 小林英司

    Journal of Reproduction and Development   51 ( Supplement )   J135 - j135   2005.8

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    DOI： 10.14882/jrds.98.0.163.0

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  • Effect of duration for in vitro maturation of recipient oocytes on nuclear remodeling and embryo development after SCNT in miniature pigs.

    Sugimura Satoshi, Wakai Takuya, Yamanaka Kenichi, Syoji Takehisa, Sasada Hiroshi, Sato Eimei

    The Journal of Reproduction and Development Supplement   98 ( 0 )   100 - 100   2005.8

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  • アクチンフィラメントを指標としたミニブタ再構築胚活性化方法の最適化

    山中 賢一, 杉村 智史, 庄司 孟央, 若井 拓哉, 佐々田 比呂志, 佐藤 英明

    日本畜産学会大会講演要旨集   105回   12 - 12   2005.8

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  • ミャンマー連邦の畜産

    田中穂積, 若井拓哉, 金川弘司

    北海道獣医師会雑誌   49 ( 7 )   13 - 17   2005.7

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  • ブタ核移植技術の現状と応用

    若井 拓哉, 佐藤 英明, 田中 穂積, 小林 英司

    Organ Biology   11 ( 2 )   169 - 169   2004.5

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  • 実験動物としての豚の現況

    田中穂積, 若井拓哉, 吉野浩之, 椎貝正太, 小林英司

    Organ Biology   11 ( 1 )   33 - 40   2004.4

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  • カフェイン添加によるMPF活性の低下抑制およびドナー細胞の条件がミニブタ再構築はいの発生に及ぼす影響

    山中賢一, 川原学, 若井拓哉, 杉村智史, 佐々田比呂志, 佐藤英明

    日本畜産学会大会講演要旨   103rd   104   2004.3

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  • カフェイン添加によるMPF活性の低下抑制およびドナー細胞の条件がミニブタ再構築胚の発生に及ぼす影響

    山中 賢一, 川原 学, 若井 拓哉, 杉村 智史, 佐々田 比呂志, 佐藤 英明

    日本畜産学会大会講演要旨集   103回   104 - 104   2004.3

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  • 声の広場 家畜豚の実験使用によるE型肝炎ウイルス対策

    田中穂積, 吉野浩之, 若井拓哉, 袴田陽二, 椎貝正太, 長尾慶和, 岡本宏明, 小林英司

    Organ Biology   10 ( 4 )   339 - 347   2003.12

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  • 譲渡犬の実験使用中止の動向とその対応に関するアンケート調査

    椎貝正太, 長尾慶和, 吉野浩之, 若井拓哉, 袴田陽二, 田中穂積, 小林英司

    Organ Biology   10 ( 4 )   321 - 329   2003.12

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  • 中国海南省の在来豚の現況と世界の実験用豚の趨勢

    田中穂積, 王希龍, 若井拓哉, 小林英司, 金川弘司

    北海道獣医師会雑誌   47 ( 12 )   440 - 443   2003.12

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  • 家畜豚の実験使用におけるE型肝炎ウイルス対策

    田中 穂積, 吉野 浩之, 若井 拓哉, 袴田 陽二, 椎貝 正太, 長尾 慶和, 岡本 宏明, 小林 英司

    Organ Biology   10 ( 4 )   339 - 347   2003.12

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  • ピギーバック方式を用いたブタ肝移植

    山内 仁, 吉野 浩之, 井上 成一朗, 清水 尚, 藤代 準, 若井 拓哉, 金子 隆志, 高橋 将文, 袴田 陽二, 田中 穂積, 小林 英司

    今日の移植   16 ( 6 )   635 - 636   2003.11

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  • Non-penetrating vascular closure staples (VCS) clipを用いたブタ部分小腸移植法

    吉野 浩之, 山内 仁, 井上 成一朗, 清水 尚, 藤代 準, 若井 拓哉, 金子 隆志, 高橋 将文, 袴田 陽二, 田中 穂積, 小林 英司

    今日の移植   16 ( 6 )   637 - 638   2003.11

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  • Activity of Maturation Promoting Factor (MPF) and distribution of mitochondria in murine in vivo and in vitro matured oocytes with different culture media from IVM.

    H Yoshida, N Aono, T Yoshida, T Wakai, H Sasada, E Sato

    FERTILITY AND STERILITY   80   S267 - S267   2003.9

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  • Activation of porcine reconstituted embryos by complex treatment

    FUCHINOUE Kohei, KAWAHARA Manabu, WAKAI Takuya, SASADA Hiroshi, KYONO Koichi, SATO Eimei

    Journal of mammalian ova research = 日本哺乳動物卵子学会誌   19 ( 2 )   S17   2002.4

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  • Combined activation of porcine oocytes injected with nuclei of fetal miniature pig cells and their subsequent development in vitro.

    K Fuchinoue, M Kawahara, T Wakai, H Sasada, K Kyono, E Sato

    BIOLOGY OF REPRODUCTION   66   238 - 239   2002

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Research Projects

  • Studies on motility control mechanism of mammalian sprematozoa using ATP imaging

    Grant number：23K11818  2023.04 - 2026.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    松浦 宏治, 森松 賢順, 若井 拓哉

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    Grant amount：\4810000 （ Direct expense: \3700000 、 Indirect expense：\1110000 ）

    １）蛍光イメージングにおける強度解析の結果から、ヒト精子内のATP量は鞭毛尾部側よりもミトコンドリアが存在している頸部の方が多いと示唆された。不動精子の場合、ATPが消費されずに産生し続ける場合があるために、蛍光観察が容易であった。運動精子は移動するために露光時間を十分に稼ぐことが困難であったことから、蛍光輝度分布と精子運動速度との関係を各動物種の精子で評価するための工夫が必要と考える。例えば、精子頭部の固定が挙げられる。阻害剤3-bromopyruvate (3BP)をヒト精液に添加し、5分後程度に観察した際には、頸部に存在する発光強度の減少が観察された。よって、本蛍光顕微観察系が本研究の目的であるATPイメージング実験に適用できると認識する。
    ２）酸素濃度を大気よりも増大した環境内にヒト精子を置いた場合、大気中の場合と比較して、ヒト精子内ATP産生量の増大が見られた。また、この場合、精子運動性、特に頭部振幅が上昇した。頸部に存在するミトコンドリアの活性が上昇し、ATP生成量が増加し、その結果頭部振幅が上昇したものと考える。
    ３）エクオリン・セレンテラジンの化学発光による細胞内カルシウム濃度評価を試みているが、試験紙上でのシグナルの分布が均一ではないために、化学発光ではなく、蛍光顕微観察中心の評価を考えている。一方、ルシフェリン・ルシフェラーゼを用いた化学発光によるATP発光を試験紙デバイスで評価する系は機能しているために、その原因を検討する。

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  • 着床前胚発生における微小核形成へのミトコンドリア機能の関与

    Grant number：23K08869  2023.04 - 2026.03

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    若井 拓哉, 杉村 智史

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    Grant amount：\4680000 （ Direct expense: \3600000 、 Indirect expense：\1080000 ）

    卵母細胞や初期胚の染色体異常は、母体の加齢に伴い増加することが広く知られている。本研究では、初期胚において染色体異数性の主要因となる微小核に着目し、その形成メカニズムの解明に取り組む。これまでの研究から、加齢雌マウスの初期胚ではミトコンドリアが染色体周囲に過剰に集積する現象が確認されている。そこで、局在が異常化したミトコンドリアの機能不全やそれに伴う活性酸素種(ROS)の増加が、染色体不安定性と微小核形成の一要因となる」と仮説を立てた。
    2023年度は、12～14か月齢のマウスから採取した加齢胚のミトコンドリア機能を解析した。ミトコンドリア膜電位をTMRM染色、また細胞内およびミトコンドリア内ROSレベルを、それぞれH2DCFDA染色とMitoSOX染色により評価した。加齢胚のミトコンドリアは、若齢胚と比較して凝集サイズの増加が観察されたが、TMRM染色による蛍光輝度に有意な差は認められなかった。一方、H2DCFDA染色による細胞内ROSレベルも加齢胚で有意な変化がみられなかったが、MitoSOX染色では凝集したミトコンドリアに顕著なROSの集積が観察された。これらの結果から、加齢に伴い顕著なミトコンドリア機能の低下はみられないものの、ミトコンドリアにおける局所的なROSの増加が起こる可能性が示唆された。
    また、着床前発生過程でのROSレベルの時空間的変化を解析するために、遺伝子コード型の蛍光プローブの使用を検討した。これまでに、酸化還元感受性GFP(roGFP)やHyper7などの異なる蛍光プローブの発現ベクターを構築した。今後はそれぞれのmRNAを合成し、初期胚における発現の観察とROSプローブとしての有用性を検討する予定である。

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  • 卵母細胞における小胞体-ミトコンドリア接触領域の生理機能解析

    Grant number：20K06627  2020.04 - 2023.03

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    若井 拓哉

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    Grant amount：\4290000 （ Direct expense: \3300000 、 Indirect expense：\990000 ）

    卵母細胞は、生涯で排卵まで至る数に限りがあり、また加齢に伴い品質低下することが広く知られている。本研究では、哺乳動物の卵母細胞の品質を制御すら細胞小器官（オルガネラ）として、ミトコンドリアと小胞体に着眼し、特に両オルガネラの接触領域（Mitochondria-associated ER membrane：MAM）の制御とその生理的な役割を明らかにすることを目的とした。
    令和3年度は、MAMの局在を高い解像度で解析するために、Split-GFPと呼ばれる分断された蛍光タンパク質を利用し、MAMの可視化を試みた。Split-GFPを発現させた受精卵では、MAMと思われる領域にGFP蛍光が確認された。また加齢卵では、ミトコンドリアの凝集に伴いMAMの凝集が観察され、両オルガネラのテザリング機能の破綻が示唆された。また本研究ではMAMの生理的機能として、ミトコンドリアと小胞体間のCa2+シグナルに着目している。正常卵を用いて、ミトコンドリアのmitochondrial calcium uniporter (MCU)およびその制御パートナーとして(Mitochondrial Calcium Uptake 1)のタンパク質発現をウエスタンブロッティング法により解析し、発現を確認した。
    また令和2年度の研究で、ミトコンドリアの動態を制御するDynamin rerated protein 1 (Drp1)を阻害した受精卵ではミトコンドリアが顕著に凝集し、胚発生停止することを明らかとしたが、発生停止要因の詳細を解明した。

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  • 不妊マウスから単離された新規原因遺伝子Adamtsl2による卵巣機能制御の解明

    Grant number：20K06470  2020.04 - 2023.03

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    辻 岳人, 若井 拓哉

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    Grant amount：\4290000 （ Direct expense: \3300000 、 Indirect expense：\990000 ）

    正常マウス卵巣におけるADAMTSL2タンパク局在を明らかにするため、４領域の異なる組換えタンパクを抗原とするウサギポリクローナル抗体を作製し、Adamtsl2遺伝子変異をもつミュータントマウスと正常マウスの卵巣およびその他組織における免疫染色を実施した。それぞれの組織において一定の陽性反応は得られるものの特異性が十分と判断できるシグナルを得ることかができなかった。さらに、in situハイブリダイゼーションによる卵巣におけるAdamtsl2遺伝の発現様式についても解析を行なった。結果として、シグナル強度は弱いものの特定の細胞に限定した発現様式を確認した。また、Adamtsl2遺伝子変異による卵巣、子宮、下垂体への影響を解析するため、RNA-seqによるミュータントマウスと正常マウスでの網羅的な遺伝子発現量の比較をおこなった。その結果、ミュータントマウスにおいてステロイドホルモン合成経路の酵素や複数のホルモンの遺伝子発現が際立って低下していることが明らかになった。一方、これまでに相互作用因子として報告されている細胞外マトリックスの遺伝子発現には差がなかった。さらに、Adamtsl2遺伝子変異によるタンパク局在への影響を解析するため、前年度作製した全長のAdamtsl2遺伝子発現ベクターおよび遺伝子変異から予想される短縮型のAdamtsl2遺伝子発現ベクターを培養細胞へ導入してタンパク局在への変化を比較した。その結果、正常な全長のADAMTSL2タンパクは細胞外での局在が確認されたが、短縮型では細胞内にのみ局在が確認され、異常タンパクとして小胞体へ取り込まれていることが予想された。

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  • Creation of mammalian oocytes with high fertility by spatiotemporal regulation of the function of mitochondria

    Grant number：18H02328  2018.04 - 2022.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    FUNAHASHI Hiroaki

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    Grant amount：\17290000 （ Direct expense: \13300000 、 Indirect expense：\3990000 ）

    The aim of this study was to generate oocytes with high developmental competence by regulating the mitochondrial (mt) DNA copy number in the ooplasm. Porcine ooplasmic mtDNA copy number did not change during in vitro maturation (IVM) and was lower in oocytes from small than medium follicles. Drp1 (mt mitogen) was reduced during early development, and its inhibition impaired the mt distribution in oocytes and early developmental competence. Overexpression of PGC1alpha (mt synthesis-related genes) resulted in increased reactive oxygen species and mtDNA copy number reduction in oocytes, and reduced IVM rate and early developmental competence. The autophagic potential of oocytes varied greatly during IVM and early development, and differed significantly by follicle diameter of origin. Attempts to introduce photosensitizers and peptide/protein molecular tools into oocytes have not resulted in significant improvement.

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  • Photo-triggered translation using caged aminoacyl-tRNA in early embryos

    Grant number：17K19202  2017.06 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Research (Exploratory)

    Ohtsuki Takashi

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    Grant amount：\6500000 （ Direct expense: \5000000 、 Indirect expense：\1500000 ）

    Light is one of the most easily manipulated external factors for spatiotemporally controlling biological events, such as protein biosynthesis. We synthesized a (7-diethylaminocoumarin-4-yl)methoxycarbonyl (DEACM)-caged aminoacyl-tRNAs (aa-tRNAs) for photocontrolling protein biosynthesis. Upon irradiation at 400-430 nm, DEACM-aa-tRNA was rapidly converted into active aa-tRNA. Translation was photoinduced when DEACM-aa-tRNAs carrying a CUA anticodon were used together with mRNAs harboring a UAG amber codon. Protein synthesis was phototriggered not only in an in vitro translation system, but also in mammalian cells. We tried to apply this strategy to photo-trigger synthesis of a fluorescent protein and PGC-1alpha in a mouse embryonic cell.

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  • Organelle dynamics during meiotic maturation of mouse oocytes

    Grant number：17K17905  2017.04 - 2020.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Young Scientists (B)

    Wakai Takuya

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    Grant amount：\4420000 （ Direct expense: \3400000 、 Indirect expense：\1020000 ）

    Ovarian ageing is characterized by quantitative and qualitative alteration of the ovarian oocyte. Mitochondria and other organelles could be the main factors determining oocyte quality adversely affected by ageing. In the present study, we focus on the organelle dynamics during oocyte maturation and fertilization. Mitochondria and endoplasmic reticulum (ER) underwent a spatial redistribution during meiotic maturation. We found organelle dynamics are co-ordinately controlled during meiotic maturation. In addition, we demonstrate that mitochondria take up Ca2+ from the ER, and increase the ATP levels of oocytes. Our results suggest organelle dynamics could underpin the successful meiotic maturation and fertilization.

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  • Cryopreservation of ovarian tissue

    Grant number：16K11089  2016.04 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Motohashi Hideyuki

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    Grant amount：\4680000 （ Direct expense: \3600000 、 Indirect expense：\1080000 ）

    The cancer treatments in young women such as anti-cancer drugs and radiation can affect fertility by several biologic systems. The purpose of this study was to develop a novel cryopreservation method for ovarian tissue. In this study, an improved vitrification for ovarian tissue resulted in survival and subsequent development in vivo and oocyte maturation in vitro after cryopreservation.

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