Molecular mechanisms of annual killifish diapause.

• Project/Area Number: 15K07074
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Developmental biology
• Research Institution: The University of Tokyo
• Principal Investigator: KUROKAWA DAISUKE 東京大学, 大学院理学系研究科(理学部), 助教 (40342779)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 一年魚 / 発生休止 / 休眠 / 細胞 / 発生 / 発生進化

Outline of Final Research Achievements

The diapause of vertebrate was analyzed using african annual killifish, Nothobranchius korthausae.
In order to investigate the function of genes involved in the diapause that expression fluctuates before and after the developmental arrest, mutants were created with CRISPR / Cas9 and the phenotype was observed.
In addition, in order to visualize the cell division cycle associated with developmental arrest, a Fluorescent Ubiquitnation-based cell cycle indicator (Fucci) transgenic line was established, and changes in cell cycle associated with the developmental arrest were observed.

Academic Significance and Societal Importance of the Research Achievements

これまで研究に用いられてこなかったアフリカ産の小型魚類Nothobranchius korthausaeを用いて、ほとんど解析が進んでいない脊椎動物の発生休止現象の解明に先鞭をつけた。また、本研究を通じて、これまで研究に用いられてこなかったN. korthausaeを、メダカやゼブラフィッシュのように簡単に実験室で飼育維持する方法や、蛍光タンパク質遺伝子を導入するトランスジェニック技術や、CRISPR/Cas9等のゲノム編集技術も確立し、新しい実験モデル生物を提案できた。

Research Products

• [Journal Article] Spatially restricted dental regeneration drives pufferfish beak development (2017)
  • Author(s): Thiery Alexandre P.、Shono Takanori、Kurokawa Daisuke、Britz Ralf、Johanson Zerina、Fraser Gareth J.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 114 Issue: 22
  • DOI: 10.1073/pnas.1702909114
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Nervous system disruption and swimming abnormality in early-hatched pufferfish (Takifugu niphobles) larvae caused by pyrene is independent of aryl hydrocarbon receptors. (2017)
  • Author(s): Itoyama Tatsuya, Kawara Moe, Fukui Makiko, Sugahara Yuki, Kurokawa Daisuke, Kawaguchi Masahumi, Kitamura Sin-Ichi, Nakayama Kei, Murakami Yasunori.
  • Journal Title: Marine Pollution Bulletin Volume: 印刷中 Issue: 2 Pages: 30174-1
  • DOI: 10.1016/j.marpolbul.2017.02.058
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Embryonic development of the grass pufferfish ( Takifugu niphobles ): From egg to larvae (2017)
  • Author(s): Gallego V.、Yoshida M.、Kurokawa D.、Asturiano J.F.、Fraser G.J.
  • Journal Title: Theriogenology Volume: 90 Pages: 191-196
  • DOI: 10.1016/j.theriogenology.2016.12.005
  • Peer Reviewed / Int'l Joint Research
• [Presentation] 一年魚の初期発生:: Molecular Mechanisms of Deep Cell Reaggregation during Early Developent of Annual Killifish Nothobranchius korthausae. (2017)
  • Author(s): 黒川大輔、吉田真明 小倉淳、相沢慎一、赤坂甲治
  • Organizer: 日本動物学会88会大会(富山)