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Development of a knock-in method and functional analysis of Kenyon cell subtypes in the mushroom bodies of the honey bee.

Research Project

Project/Area Number 20K15839
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 44050:Animal physiological chemistry, physiology and behavioral biology-related
Research InstitutionThe University of Tokyo

Principal Investigator

Kohno Hiroki  東京大学, 大学院理学系研究科(理学部), 助教 (60846773)

Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2022: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2021: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Keywordsセイヨウミツバチ / 社会性行動 / ゲノム編集 / キノコ体 / ケニヨン細胞 / ノックイン
Outline of Research at the Start

ミツバチの社会性行動を制御する神経基盤として、脳高次中枢キノコ体に着目した研究が多く行われてきた。ミツバチのキノコ体を構成するケニヨン細胞にはサブタイプが存在し、異なる行動の制御に関わることが示唆されている。本研究では、ミツバチにおいてゲノム編集によるノックイン法を実現することで、各サブタイプ選択的な遺伝子発現操作を行い、各サブタイプの投射パターン、網羅的な遺伝子発現プロファイル、そして行動制御における機能を同定する。ケニヨン細胞サブタイプはハチ目昆虫の行動の複雑化に伴って増加したことが示唆されており、本研究により行動進化に寄与する神経機構の理解に繋がることが期待される。

Outline of Final Research Achievements

The mushroom bodies, a higher center of the insect brain, have been studied as a brain region related to social behaviors in the honey bee. The honey bee mushroom bodies are composed of several Kenyon cell subtypes, each of which has distinct gene expression profiles. However, the function of each subtype in behavioral regulation remains to be elucidated. This study tried to establish a knock-in method using CRISPR/Cas9 to realize cell type-specific gene manipulation. I also analyzed functions of the gene which is selectively expressed in a certain subtype by producing its knockout mutants, and identified projection patterns and comprehensive gene expression profiles for each Kenyon cell subtype to reveal some of the functional and physiological characteristics of each subtype.

Academic Significance and Societal Importance of the Research Achievements

ミツバチは社会性行動研究のモデル生物として研究されてきたが、その行動特性から遺伝子操作を行うことが長年難しく、いまだ細胞種特異的な遺伝子発現制御がなされた例は無い。本研究は予備的ながら初めてノックイン法の有効性を示した。加えて、継代によるノックアウト働き蜂の作出に初めて成功しており、ミツバチの行動遺伝学に大きく貢献する成果と考える。また、行動進化と関連することが示唆されているキノコ体の生理特性の一端を解明したことで、今後社会性段階の異なるハチ目昆虫との比較解析により、社会性と関連する脳基盤の発見に繋がる可能性がある。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • Research Products

    (11 results)

All 2023 2022 2021 2020 Other

All Journal Article (4 results) (of which Peer Reviewed: 3 results,  Open Access: 3 results) Presentation (6 results) (of which Int'l Joint Research: 2 results) Remarks (1 results)

  • [Journal Article] Evolutionary dynamics of mushroom body Kenyon cell types in hymenopteran brains from multi-functional type to functionally specialized types.2023

    • Author(s)
      Takayoshi Kuwabara, Hiroki Kohno, Masatsugu Hatakeyama, Takeo Kubo
    • Journal Title

      Science Advances

      Volume: Add4210 Issue: 18

    • DOI

      10.1126/sciadv.add4201

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Mblk-1/E93, an ecdysone related-transcription factor, targets synaptic plasticity-related genes in the honey bee mushroom bodies2022

    • Author(s)
      Matsumura Yasuhiro、To Taiko Kim、Kunieda Takekazu、Kohno Hiroki、Kakutani Tetsuji、Kubo Takeo
    • Journal Title

      Scientific Reports

      Volume: 12 Issue: 1 Pages: 21367-21367

    • DOI

      10.1038/s41598-022-23329-z

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Developmental stage-specific distribution and phosphorylation of Mblk-1, a transcription factor involved in ecdysteroid-signaling in the honey bee brain2020

    • Author(s)
      Kumagai Hitomi、Kunieda Takekazu、Nakamura Korefumi、Matsumura Yasuhiro、Namiki Manami、Kohno Hiroki、Kubo Takeo
    • Journal Title

      Scientific Reports

      Volume: 10 Issue: 1 Pages: 8735-8735

    • DOI

      10.1038/s41598-020-65327-z

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] ミツバチ脳における脱皮ホルモンシグナル関連転写因子Mblk-1の発現解析2020

    • Author(s)
      松村泰宏、河野大輝、久保健雄
    • Journal Title

      昆虫と自然

      Volume: 56 Pages: 30-33

    • NAID

      40022469603

    • Related Report
      2020 Research-status Report
  • [Presentation] Identification of the pupal stages at which three Kenyon cell subtypes are generated in the mushroom bodies of the honey bee2022

    • Author(s)
      Hiroki Kohno, Takeo Kubo
    • Organizer
      日本発生生物学会第55回大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Functional analyses of mKast by producing knocked out worker honey bees using CRISPR/Cas92022

    • Author(s)
      Hiroki Kohno, Takeo Kubo
    • Organizer
      IUSSI2022 19th Congress of the International Union for the Study of Social Insects
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Analysis and exploration of Kenyon cells increased with behavioral evolution in hymenopteran insects2022

    • Author(s)
      Takayoshi Kuwabara, Hiroki Kohno, Masatsugu Hatakeyama, Takeo Kubo
    • Organizer
      IUSSI2022 19th Congress of the International Union for the Study of Social Insects
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ミツバチのダンスコミュニケーションに関わる神経細胞種の探索 ~ミツバチとマルハナバチの脳高次中枢の比較scRNA-seq解析~2022

    • Author(s)
      桑原嵩佳、河野大輝、久保健雄
    • Organizer
      シングルセルゲノミクス研究会2022
    • Related Report
      2022 Annual Research Report
  • [Presentation] 行動を司る脳進化の謎へ切り込む ~シンプルな脳を持つハチ目昆虫の種間比較解析より~2022

    • Author(s)
      桑原嵩佳、河野大輝、畠山正統、久保健雄
    • Organizer
      第7回ユニーク会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Functional analyses of mKast, which is preferentially expressed in middle-type Kenyon cells in mushroom bodies of the honey bee, by producing knocked-out workers2021

    • Author(s)
      Hiroki Kohno, Takeo Kubo
    • Organizer
      日本比較生理生化学会 第43回札幌オンライン大会
    • Related Report
      2021 Research-status Report
  • [Remarks] ハチ目における脳神経細胞の進化動態の解明 ――多機能型から機能特化型へ――

    • URL

      https://www.s.u-tokyo.ac.jp/ja/press/2023/8416/

    • Related Report
      2022 Annual Research Report

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Published: 2020-04-28   Modified: 2024-01-30  

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