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Maintenance of trophectoderm in bovine vitrified embryos based on elucidation of regulatory mechanism of early embryonic differentiation

Research Project

Project/Area Number 21H02336
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 42010:Animal production science-related
Research InstitutionHokkaido University

Principal Investigator

Kawahara Manabu  北海道大学, 農学研究院, 准教授 (70468700)

Co-Investigator(Kenkyū-buntansha) 柳川 洋二郎  北海道大学, 獣医学研究院, 准教授 (20609656)
唄 花子  北海道大学, 農学研究院, 助教 (60775443)
Project Period (FY) 2021-04-01 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000)
Fiscal Year 2023: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2022: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2021: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Keywords初期胚発生 / マウス / ウシ / ガラス化保存 / 細胞分化 / 胚盤胞期胚 / Hippo経路 / Nf2 / Yap1 / 栄養膜細胞 / 初期胚 / 個体発生 / 動物生産
Outline of Research at the Start

胚盤胞期胚の胎子側および胎盤側(栄養外胚葉)への細胞分化機構を解明することは、生物学的にも動物生産の効率化のためにも不可欠である。しかし、最も解析が進んでいるマウス胚ですら、その全容は解析途上で、実験手段が限られるウシ胚では更に知見に乏しい。本研究では、ウシ胚盤胞期胚の栄養外胚葉細胞における分化維持遺伝子CDX2発現に必要なYAP1タンパク質の局在制御に及ぼす細胞膜/細胞骨格関連タンパク質NF2の役割をマウスおよびウシ胚を用いて明らかしつつ、得られた知見に基づいてウシガラス化胚の発生能力向上を目指す。

Outline of Final Research Achievements

This study showed that the localization of the scaffold protein NF2 plays an important role in regulating the subcellular localization of YAP1, which determines TE cell differentiation in mouse and bovine, and that immersion in hypertonic solution that is a routine part of embryo vitrification procedure alters YAP1 localization. These findings have developmental biological significance in clarifying part of the regulatory mechanism of differentiation of mammalian embryos, indicating that the maintenance of subcellular localization of YAP1 could be impaired by vitrification. These outcomes are expected to contribute to the elucidation of new molecular mechanisms in the regulation of mammalian embryo development and to the refinement of vitrification preservation techniques that contribute to the improvement and proliferation of bovine embryos.

Academic Significance and Societal Importance of the Research Achievements

細胞の極性に伴う細胞分化機構は古くから知られているものの、分子レベルでの理解は進んでいなかった。また正常に分化した初期胚の作製は、主要なウシの繁殖方法の一つである受精卵移植技術の根幹となる発生工学技術でもある。さらに、ウシ生産のために実質的に受精卵移植技術を活用していくためには受精卵の低温下での保存技術の進展が欠かせない。本研究で得られた知見をもとに、哺乳類胚の発生制御における新たな分子機構の解明が進むとともに、ウシ胚改良増殖に不可欠なガラス化保存技術の洗練化につながることが期待される。

Report

(3 results)
  • 2023 Final Research Report ( PDF )
  • 2022 Annual Research Report
  • 2021 Annual Research Report
  • Research Products

    (26 results)

All 2023 2022 2021

All Journal Article (14 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 14 results,  Open Access: 2 results) Presentation (11 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Synergistic effect of standardized extract of Asparagus officinalis stem and heat shock on progesterone synthesis with lipid droplets and mitochondrial function in bovine granulosa cells2023

    • Author(s)
      Ho Khoi Thieu, Balboula Ahmed Zaky, Homma Kohei, Takanari Jun, Bai Hanako, Kawahara Manabu, Thi Kim Nguyen Khang, Takahashi Masashi
    • Journal Title

      The Journal of Steroid Biochemistry and Molecular Biology

      Volume: 225 Pages: 106181-106181

    • DOI

      10.1016/j.jsbmb.2022.106181

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Identification of menaquinone-4 (vitamin K2) target genes in bovine endometrial epithelial cells in?vitro2023

    • Author(s)
      Bai Hanako, Kawahara Manabu, Takahashi Masashi
    • Journal Title

      Theriogenology

      Volume: 198 Pages: 183-193

    • DOI

      10.1016/j.theriogenology.2022.12.033

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Generation of viable calves derived from developmentally mature blastocysts produced by on-gel culture2022

    • Author(s)
      SAITO Shun、AKIZAWA Hiroki、FURUKAWA Eri、YANAGAWA Yojiro、BAI Hanako、TAKAHASHI Masashi、KAWAHARA Manabu
    • Journal Title

      Journal of Reproduction and Development

      Volume: 68 Issue: 5 Pages: 330-334

    • DOI

      10.1262/jrd.2022-054

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Zona pellucida removal by acid Tyrode’s solution affects pre- and post-implantation development and gene expression in mouse embryos2022

    • Author(s)
      Fan Weihong、Huang Tengda、Wu Tian、Bai Hanako、Kawahara Manabu、Takahashi Masashi
    • Journal Title

      Biology of Reproduction

      Volume: -

    • DOI

      10.1093/biolre/ioac155

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Cattle production by intracytoplasmic sperm injection into oocytes vitrified after ovum pick-up2022

    • Author(s)
      Kagawa Shinjiro、Hiraizumi Shingo、Bai Hanako、Takahashi Masashi、Kawahara Manabu
    • Journal Title

      Theriogenology

      Volume: 185 Pages: 121-126

    • DOI

      10.1016/j.theriogenology.2022.03.022

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Environmental factors affecting the conception rates of nulliparous and primiparous dairy cattle2022

    • Author(s)
      Ukita Haruka、Yamazaki Takeshi、Yamaguchi Satoshi、Abe Hayato、Baba Toshimi、Bai Hanako、Takahashi Masashi、Kawahara Manabu
    • Journal Title

      Journal of Dairy Science

      Volume: -

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Requirement for expression of WW domain containing transcription regulator 1 in bovine trophectoderm development2021

    • Author(s)
      Saito Shun、Yamamura Shota、Kohri Nanami、Bai Hanako、Takahashi Masashi、Kawahara Manabu
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 555 Pages: 140-146

    • DOI

      10.1016/j.bbrc.2021.03.112

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Loop-mediated isothermal amplification (LAMP) and machine learning application for early pregnancy detection using bovine vaginal mucosal membrane2021

    • Author(s)
      Kunii Hiroki, Kubo Tomoaki, Asaoka Natsuki, Balboula Ahmed Z, Hamaguchi Yu, Shimasaki Tomoya, Bai Hanako, Kawahara Manabu, Kobayashi Hisato, Ogawa Hidehiko, Takahashi Masashi
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 569 Pages: 179-186

    • DOI

      10.1016/j.bbrc.2021.07.015

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Heat stress induces oxidative stress and activates the KEAP1-NFE2L2-ARE pathway in bovine endometrial epithelial cells2021

    • Author(s)
      Murata Hirona, Kunii Hiroki ,Kusama Kazuya, Sakurai Toshihiro, Bai Hanako、Kawahara Manabu、Takahashi Masashi
    • Journal Title

      Biology of Reproduction

      Volume: 105 Issue: 5 Pages: 1114-1125

    • DOI

      10.1093/biolre/ioab143

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mitochondrial maturation in the trophectoderm and inner cell mass regions of bovine blastocysts2021

    • Author(s)
      Hayashi Yoshihiro, Saito Shun, Bai Hanako, Takahashi Masashi, Kawahara Manabu
    • Journal Title

      Theriogenology

      Volume: 175 Pages: 69-76

    • DOI

      10.1016/j.theriogenology.2021.08.038

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The use of a two-step removal protocol and optimized culture conditions improve development and quality of zona free mouse embryos2021

    • Author(s)
      Fan Weihong, Homma Misato, Xu Renliang, Kunii Hiroki, Bai Hanako, Kawahara Manabu, Kawaguchi Toshikazu, Takahashi Masashi
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 577 Pages: 116-123

    • DOI

      10.1016/j.bbrc.2021.09.013

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A standardized extract of Asparagus officinalis stem improves HSP70-mediated redox balance and cell functions in bovine cumulus-granulosa cells2021

    • Author(s)
      Ho Khoi Thieu, Homma Kohei, Takanari Jun, Bai Hanako, Kawahara Manabu, Nguyen Khang Thi Kim, Takahashi Masashi
    • Journal Title

      Scientific Reports

      Volume: 11 Issue: 1 Pages: 18175-18175

    • DOI

      10.1038/s41598-021-97632-6

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Deciphering two rounds of cell lineage segregations during bovine preimplantation development2021

    • Author(s)
      Akizawa Hiroki, Saito Shun, Kohri Nanami, Furukawa Eri, Hayashi Yoshihiro, Bai Hanako, Nagano Masashi, Yanagawa Yojiro, Tsukahara Hayato, Takahashi Masashi, Kagawa Shinjiro, Kawahara‐Miki Ryouka, Kobayashi Hisato、Kono Tomohiro、Kawahara Manabu
    • Journal Title

      The FASEB Journal

      Volume: 35 Issue: 10

    • DOI

      10.1096/fj.202002762rr

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Cell-cycle dependent GATA2 subcellular localization in mouse 2-cell embryos2021

    • Author(s)
      Komatsu Masaya, Tsukahara Hayato, Bai Hanako, Takahashi Masashi, Wakai Takuya, Kawahara Manabu
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 584 Pages: 1-6

    • DOI

      10.1016/j.bbrc.2021.10.077

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Presentation] ウシ子宮内膜上皮細胞における暑熱負荷培養時のNFE2L2の標的遺伝子の探索.2022

    • Author(s)
      唄 花子, 川原 学, 高橋 昌志.
    • Organizer
      日本繁殖生物学会 (東京都 東京大学)
    • Related Report
      2022 Annual Research Report
  • [Presentation] ウシ卵子成熟および発生における熱ショック非依存的なHSP70発現誘導と酸化ストレスに及ぼすEASの影響.2022

    • Author(s)
      楠野莉奈子, Ho Khoi, 本間康平, 高成準, 唄花子, 川原学, 高橋昌志.
    • Organizer
      日本畜産学会 (神奈川県 東京農業大学)
    • Related Report
      2022 Annual Research Report
  • [Presentation] ウシ初期胚発生の研究.2022

    • Author(s)
      川原 学
    • Organizer
      日本繁殖生物学会 (オンライン)
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] 内部細胞塊におけるウシYAP1の局在2022

    • Author(s)
      齋藤 隼, 加川 真二郎, 川原 学.
    • Organizer
      日本分子生物学会 (千葉県 幕張メッセ)
    • Related Report
      2022 Annual Research Report
  • [Presentation] Gene expression changes in trophoblast cells after the blastocyst stage in cattle.2022

    • Author(s)
      Akizawa H., Bai H., Takahashi M. and Kawahara M.
    • Organizer
      International Embryo Technology Society 48th Annual Conference
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ホルスタイン種初産牛の人工授精受胎率に及ぼす環境要因の解析.2021

    • Author(s)
      宇喜多遥, 山崎武志, 山口諭, 阿部隼人, 馬場俊見, 唄花子, 高橋昌志, 川原学.
    • Organizer
      日本畜産学会 (宮城県 東北大学)
    • Related Report
      2021 Annual Research Report
  • [Presentation] Effect of two-step combined zona pellucida removal on developmental competence of mouse embryos.2021

    • Author(s)
      Weihong Fan, Hanako Bai, Manabu Kawahara, Masashi Takahashi.
    • Organizer
      日本畜産学会 (宮城県 東北大学)
    • Related Report
      2021 Annual Research Report
  • [Presentation] Effect of EAS on post freezing viability of heat stressed bovine cumulus granulosa cells in relation with redox balance.2021

    • Author(s)
      Khoi Thieu Ho, Kohei Homma, Jun Takanari, Hanako Bai, Manabu Kawahara, Kim Khang Nguyen Thi, Masashi Takahashi.
    • Organizer
      日本畜産学会 (宮城県 東北大学)
    • Related Report
      2021 Annual Research Report
  • [Presentation] Effect of EAS on P4 production pathway of bovine CG cells in relation with HSP70.2021

    • Author(s)
      Khoi Thieu HO, Kohei HOMMA, Jun TAKANARI, Hanako BAI, Manabu KAWAHARA, Kim Khang NGUYEN THI, Masashi TAKAHASHI.
    • Organizer
      日本繁殖生物学会 (京都府 京都大学)
    • Related Report
      2021 Annual Research Report
  • [Presentation] 腟底部発現変動遺伝子を用いたRT-LAMP法および機械学習によるウシ簡易迅速妊娠判定モデルの検討.2021

    • Author(s)
      國井 宏樹,窪 友瑛,浅岡 那月,嶋崎 知哉,唄 花子,川原 学,高橋 昌志.
    • Organizer
      日本繁殖生物学会 (京都府 京都大学)
    • Related Report
      2021 Annual Research Report
  • [Presentation] ウシ子宮内膜上皮細胞における暑熱負荷培養時の酸化ストレス応答経路の検証.2021

    • Author(s)
      唄 花子,國井 宏樹,川原 学,高橋 昌志.
    • Organizer
      日本繁殖生物学会 (京都府 京都大学)
    • Related Report
      2021 Annual Research Report
  • [Patent(Industrial Property Rights)] ウシ胚の体外培養方法、ウシ胚、及びウシの生産方法2022

    • Inventor(s)
      川原 学、秋沢 宏紀、高橋 昌志、唄 花子
    • Industrial Property Rights Holder
      川原 学、秋沢 宏紀、高橋 昌志、唄 花子
    • Industrial Property Rights Type
      特許
    • Filing Date
      2022
    • Related Report
      2022 Annual Research Report

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Published: 2021-04-28   Modified: 2025-01-30  

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