The analysis of the development and the function of cerebellar neural circuitry by using zebrafish

• Project/Area Number: 25440104
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Developmental biology
• Research Institution: Nagoya University
• Principal Investigator: Shimizu Takashi 名古屋大学, 生物機能開発利用研究センター, 准教授 (50324760)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 小脳 / 神経発生 / ゼブラフィッシュ / 恐怖条件付け学習 / 顆粒細胞 / 条件付け恐怖学習 / 神経 / 細胞分化 / 細胞分裂 / 細胞増殖

Outline of Final Research Achievements

The cerebellum plays an important role for motor learning and classical fear conditioning. To understand the genes that control the generation of cerebellar granule cells and their axogenesis, I investigated two zebrafish mutants. The cranio facial development protein 1 gene controlled granule cell defferentiation by switching proliferation state to differentiation state in the precursors. The component of beaement-membrane, type IV collagens, regulated the axogenesis. I established a method for the classical fear conditioning paradigm by using zebrafish larva. The 20 days zebrafish larvae acquired the conditioned fear responses. Inhibition of granule-cell transmission impaired the recovery from conditioned responses. I found conditioning-dependent changes in granule cell encoding. There were two types of conditioning-associated neurons in the cerebellum. These results indicate that granule cells control the recovery from conditioned fear responses in zebrafish.

Research Products

• [Journal Article] Gene expression profiling of granule cells and Purkinje cells in the zebrafish cerebellum. (2017)
  • Author(s): Takeuchi M, Yamaguchi S, Sakakibara Y, Hayashi T, Matsuda K, Hara Y, Tanegashima C, Shimizu T, Kuraku S, Hibi M.
  • Journal Title: Journal of Comparative Neurology Volume: 525 Issue: 7 Pages: 1558-1585
  • DOI: 10.1002/cne.24114
  • NAID: 120006369328
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Evolutionary mechanisms that generate morphology and neural-circuit diversity of the cerebellum. (2017)
  • Author(s): Hibi, M., Matsuda, K., Takeuchi, M., Shimizu, T., and Murakami, Y.
  • Journal Title: Dev. Growth Differ. Volume: in press
  • NAID: 120006406681
• [Journal Article] Responses of cerebellar Purkinje cells during fictive optomotor behavior in larval zebrafish. (2016)
  • Author(s): Scalise K, Shimizu T, Hibi M, Sawtell NB.
  • Journal Title: Journal of Neurophysiology Volume: 116 Issue: 5 Pages: 2067-2080
  • DOI: 10.1152/jn.00042.2016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] ype IV collagen controls the axogenesis of cereberr granule cells by regulating basement membrane integrity in zebrafish. (2015)
  • Author(s): Takeuchi, M., Yamaguchi, S., Yonemura, S., Kakiguchi, K., Sato, Y., Higashiyama, T., Shimizu, T., and, Hibi, M.
  • Journal Title: PLoS genet. Volume: 11 Issue: 10 Pages: e1005587-e1005587
  • DOI: 10.1371/journal.pgen.1005587
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Establishment of Gal4 transgenic zebrafish lines for analysis of development of cerebellar neural circuitry (2015)
  • Author(s): Takeuchi, M., Matsuda, K., Yamaguchi, S., Asakawa, K., Miyasaka, N., Lal, P., Yoshihara, Y., Koga, A., Kawakami, K., Shimizu, T., and Hibi, M.
  • Journal Title: Developmental Biology Volume: 397 Issue: 1 Pages: 1-17
  • DOI: 10.1016/j.ydbio.2014.09.030
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] ゼブラフィッシュ恐怖条件付け学習における小脳の役割 (2016)
  • Author(s): 清水貴史
  • Organizer: 日本発生生物学会 秋季シンポジウム
  • Place of Presentation: 三島
  • Year and Date: 2016-10-19
• [Presentation] Roles of zebrafish cerebellar neurons in classical fear conditioning. (2016)
  • Author(s): Matsuda, K., Yoshida, M., Kawakami, K., Hibi, M., Shimizu, T.
  • Organizer: 第22回小型魚類研究会
  • Place of Presentation: 岡崎
  • Year and Date: 2016-08-20
• [Presentation] Gene expression profiling of granule cells and Purkinje cells in zebrafish cerebellum. (2016)
  • Author(s): Takeuchi, M., Yamaguchi, S., Sakakibara, Y., Hayashi, T., Matsuda, K., Hara, Y., Hayashi, T., Kuraku, S., Shimizu, T., and Hibi, M.
  • Organizer: 第22回小型魚類研究会
  • Place of Presentation: 岡崎
  • Year and Date: 2016-08-20
• [Presentation] Roles of Ptf1a and Gsx2 in development of Purkinje cells and inferior olive nucleus neurons in zebrafish. (2016)
  • Author(s): Ito, T., Takeuchi, M., Sakagami, M., Shimizu, T., and Hibi, M.
  • Organizer: 第22回小型魚類研究会
  • Place of Presentation: 岡崎
  • Year and Date: 2016-08-20
• [Presentation] Reelin signaling controls the layer formation of cerebellum and optic tectum in zebrafish. (2016)
  • Author(s): Nimura, T., Di, Donato V., Hayashi, T., Takeuchi, M., Shimizu, T., Del Bene, F., and Hibi, M.
  • Organizer: 第22回小型魚類研究会
  • Place of Presentation: 岡崎
  • Year and Date: 2016-08-20
• [Presentation] Zebrafish cerebellar neural circuitry is involved in the classical fear conditioning. (2016)
  • Author(s): Matsuda, K., Yoshida, M., Kawakami, K., Hibi, M., Shimizu, T.
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2016-07-20
• [Presentation] Basement membrane stability is important for the axogenesis of cerebellar granule cells in zebrafish. (2015)
  • Author(s): Takeuchi, M., Yamaguchi, S., Hayashi, T., Sakakibara, Y., Matsuda, K., Hara, Y., Hayashi T., Kuraku, S., Shimizu, T., and Hibi, M.
  • Organizer: 第21回小型魚類研究会
  • Place of Presentation: 吹田
  • Year and Date: 2015-09-19
• [Presentation] Transcriptome analysis of zebrafish cerebellar Purkinje cells and mormyrid fish cerebellum. (2015)
  • Author(s): Sakakibara, Y.*, Matsuda, K.*, Hayashi, T., Yamaguchi, S., Takeuchi, M., Ito, T. , Hara, Y., Hayashi T., Kuraku, S., Shimizu, T., and Hibi, M.
  • Organizer: 第21回小型魚類研究会
  • Place of Presentation: 吹田
  • Year and Date: 2015-09-19
• [Presentation] Zebrafish cerebellum is involved in the classical fear conditioning. (2015)
  • Author(s): Matsuda, K., Yoshida, M., Asakawa, K., Nakai, J., Kawakami, K., Hibi, M., and Shimizu, T.
  • Organizer: 第21回小型魚類研究会
  • Place of Presentation: 吹田
  • Year and Date: 2015-09-19
• [Presentation] Roles of zebrafish cerebellar neural circuitry in classical fear conditioning. (2015)
  • Author(s): Matsuda, K., Yoshida, M., Asakawa, K., Ohkura, M., Nakai, J., Kawakami, K., Hibi, M., and Shimizu, T.
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-07-28
• [Presentation] nuclear protein Cfdp1 regulates differentiateon of granule cells in zebrafish cerebellum. (2015)
  • Author(s): Shimizu, T., Kani, S., Bae, Y.K., Sun, X. and Hibi, M.
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-07-28
• [Presentation] Transcriptome analysis of granule cells and Purkinje cells in zebrafish cerebellum. (2015)
  • Author(s): Takeuchi, M., Yamaguchi, S., Hayashi, T., Sakakibara, Y., Matsuda, K., Hara, Y.,Hayashi, T., Kuraku, S., Shimizu, T., and Hibi, M.
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-07-28
• [Presentation] Evolution and development of cerebellar neural circuitry in teleosts. (2015)
  • Author(s): Hibi, M., Takeuchi, M., Yamaguchi, S., Matsuda, K., Hayashi, T., Hara, Y., Hayashi, T., Sakakibara, Y., Yoshida, M., Kuraku, S., and Shimizu, T.
  • Organizer: 第48回日本発生生物学会大会
  • Place of Presentation: つくば
  • Year and Date: 2015-06-02
• [Presentation] Integrity of basement membrane is important for axogenesis of cerebellar granule cells. (2015)
  • Author(s): Takeuchi, M., Yamaguchi, S., Hayashi, T., Yonemura, S., Hara, Y., Hayashi T., Kuraku, S., Shimizu, T., and Hibi, M.
  • Organizer: 第48回日本発生生物学会大会
  • Place of Presentation: つくば
  • Year and Date: 2015-06-02
• [Presentation] A nuclear protein Cfdp1 regulates differentiateon of granule cells in zebrafish cerebellum. (2015)
  • Author(s): Shimizu, T., Kani, S., Bae, Y.K., Sun, X., and Hibi, M.
  • Organizer: 第48回日本発生生物学会大会
  • Place of Presentation: つくば
  • Year and Date: 2015-06-02
• [Presentation] Visualizing development and morphology of cerebellar granule axons in zebrafish. (2014)
  • Author(s): Yamaguchi, S., Takeuchi, M., Hayashi, T., Asakawa, K., Hara, Y., Hayashi, T, Kuraku, H., Kawakami, K., Shimizu, T., and Hibi, M.
  • Organizer: 第20回小型魚類研究会
  • Place of Presentation: 東京
  • Year and Date: 2014-09-20 – 2014-09-21
• [Presentation] Roles of zebrafish cerebellar neural circuitry in the classical fear conditioning. (2014)
  • Author(s): Matsuda, K., Yoshida, M., Takeuchi, M., Asakawa, K., Ohkura, M., Miyasaka, N., Yoshihara, Y., Nakai, J., Kawakami, K., Hibi, M., and Shimizu, T.
  • Organizer: 第20回小型魚類研究会
  • Place of Presentation: 東京
  • Year and Date: 2014-09-20 – 2014-09-21
• [Presentation] Precise assembly of the basement membrane is important for the axogenesis of cerebellar granule cells. (2014)
  • Author(s): Takeuchi, Yamaguchi, S., Yonemura, S., Asakawa, K., Kawakami, K., Takada, S., Shimizu, T., and Hibi, M.
  • Organizer: 第20回小型魚類研究会
  • Place of Presentation: 東京
  • Year and Date: 2014-09-20 – 2014-09-21
• [Presentation] Basement membrane structure is important for axogenesis of cerebellar granule cells. (2014)
  • Author(s): Takeuchi, M., Yonemura, S., Asakawa, K., Kawakami, K., Takada, S., Kuraku, S., Hayashi, T., Yamaguchi, S., Shimizu, T., and Hibi, M.
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] Control of axon guidance of cerebellar granule cells by the basement membrane. (2014)
  • Author(s): Takeuchi, M., Yonemura, S., Asakawa, K., Kawakami, K., Takada, S., Kuraku, S., Hayashi, T., Yamaguchi, S.*, Shimizu, T., and Hibi, M.
  • Organizer: 11th International Conference on Zebrafish Development and Genetics.
  • Place of Presentation: Wisconsin, USA
  • Year and Date: 2014-06-24 – 2014-06-28
• [Presentation] issection of cerebellar neural circuitry with zebrafish Gal4 gene/enhancer trap lines. (2014)
  • Author(s): Matsuda, K., Takeuchi, M., Asakawa, K., Ohkura, M., Miyasaka, N., Yoshihara, Y., Hayashi, T., Kuraku, S., Kawakami, K., Hibi, M., and Shimizu, T.
  • Organizer: 第47回日本発生生物学会大会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-27 – 2014-05-30
• [Presentation] Precise axogenesis of the cerebellar granule cells requires interaction between their axons and the basement membrane. (2014)
  • Author(s): Takeuchi, M., Yonemura, S., Asakwa, K., Kawakami, K., Takada, S., Shimizu, T., and Hibi, M.
  • Organizer: 第47回日本発生生物学会大会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-27 – 2014-05-30
• [Presentation] Moving microtubule-mediated dorsal determination. (2013)
  • Author(s): Hino, H., Seki, R., Shimizu, T., Pelegri, F., and Hibi, M.
  • Organizer: 第19回小型魚類研究会
  • Place of Presentation: 仙台
• [Presentation] Functional analysis of zebrafish cerebellar neural circuitry in classical conditioning. (2013)
  • Author(s): Matsuda, K., Takeuchi, M., Asakawa, K., Yoshida, M., Ohkura, M., Miyasaka, N., Yoshihara, Y., Nakai, J., Kawakami, K., Hibi, M., and Shimizu, T.
  • Organizer: 第19回小型魚類研究会
  • Place of Presentation: 仙台
• [Presentation] Basement membrane-dependent control of axogenesis of cerebellar granule cells. (2013)
  • Author(s): Takeuchi, M., Yonemura, S., Asakawa, K., Kawakami, K., Takada, S., Shimizu, T., and Hibi, M.
  • Organizer: 第19回小型魚類研究会
  • Place of Presentation: 仙台
• [Presentation] Development of cerebellar granule cell axons in zebrafish. (2013)
  • Author(s): Yamaguchi, S., Takeuchi, M., Asakawa, K., Kawakami, K., Shimizu, T., and Hibi, M.
  • Organizer: 第46回日本発生生物学会大会
  • Place of Presentation: 松江
• [Presentation] Involvement of microtubule-dependent transport of Wnt signaling components in zebrafish dorsal determination. (2013)
  • Author(s): Hino, H., Seki, R., Shimizu, T., Long, T.D., Sampath, K., and Hibi, M.
  • Organizer: 第46回日本発生生物学会大会
  • Place of Presentation: 松江
• [Presentation] Basement membrane integrity is required for axogenesis of cerebellar granule cells. (2013)
  • Author(s): Takeuchi, M., Yonemura, S., Asakawa, K., Kawakami, K., Takada, S., Shimizu, T., and Hibi, M.
  • Organizer: 第46回日本発生生物学会大会
  • Place of Presentation: 松江
• [Remarks] 名古屋大学生物機能開発利用研究センター動物器官機能学ホームページ
  • URL: http://bbc.agr.nagoya-u.ac.jp/~junkei/
• [Remarks] IV型コラーゲンは基底膜の整合性を保つことで小脳顆粒細胞の軸索形成を制御している
  • URL: http://www.bio.nagoya-u.ac.jp/paper/2016-26/08.html
• [Remarks] 名古屋大学生物機能開発利用研究センター動物器官機能研究分野
  • URL: http://bbc.agr.nagoya-u.ac.jp/~junkei/index.html
• [Remarks] 名古屋大学生物機能開発利用研究センター動物器官機能研究分野
  • URL: http://bbc.agr.nagoya-u.ac.jp/~junkei/index.html