Establishment of recombinant rabies virus-mediated retrograde trancing of neural circuits in zebrafish

• Project/Area Number: 15K14523
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Developmental biology
• Research Institution: Nagoya University
• Principal Investigator: HIBI MASAHIKO 名古屋大学, 生物機能開発利用研究センター, 教授 (40273627)
• Co-Investigator(Renkei-kenkyūsha): OSAKADA FUMITAKA 名古屋大学, 創薬科学研究科, 准教授 (60455334)
• Research Collaborator: DOHAKU RYUJI 名古屋大学, 理学研究科
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 逆行性トレーシング / 狂犬病ウイルス / 神経回路 / 神経解剖学 / 小脳 / 苔状線維 / 登上線維 / ゼブラフィッシュ / 逆光性トレーシング

Outline of Final Research Achievements

Rabies virus (RV) infects neurons through its glycoprotein (G). Recombinant RV that contains the GFP gene instead of the G gene and is pseudotyped with the retroviral envelope protein (EnvA) was developed for neural circuit tracing. The pseudotyped RV can infect neurons that express TVA (the EnvA receptor)-mCherry. If the G is provided in the same cells, the RV particles are generated and transmitted mono-synaptically to input neurons. We generated transgenic lines that express TVA-mCherry and G in Purkinje cells (PCs) or granule cells (GCs). When PCs were infected with the pseudotyped RV, GFP was detected in GCs and neurons in the inferior olive nuclei. When GCs were infected with the virus, GFP was detected in neurons in various brain regions that receive inputs from the visual, vestibular, lateral line sensory systems, or telencephalon. Our findings validated the RV-mediated retrograde tracing in zebrafish and identified input neurons for GCs in zebrafish.

Research Products

• [Int'l Joint Research] Fred Hutchinson Cancer Research Center(米国)
• [Journal Article] Multiple zebrafish atoh1 genes specify a diversity of neuronal types in the zebrafish cerebellum (2018)
  • Author(s): Kidwell Chelsea U.、Su Chen-Ying、Hibi Masahiko、Moens Cecilia B.
  • Journal Title: Developmental Biology Volume: 印刷中 Issue: 1 Pages: 44-56
  • DOI: 10.1016/j.ydbio.2018.03.004
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Roles of maternal wnt8a transcripts in axis formation in zebrafish (2018)
  • Author(s): Hino Hiromu、Nakanishi Akiko、Seki Ryoko、Aoki Tsubasa、Yamaha Etsuro、Kawahara Atsuo、Shimizu Takashi、Hibi Masahiko
  • Journal Title: Developmental Biology Volume: 434 Issue: 1 Pages: 96-107
  • DOI: 10.1016/j.ydbio.2017.11.016
  • NAID: 120006808763
  • Peer Reviewed
• [Journal Article] Granule cells control recovery from classical conditioned fear responses in the zebrafish cerebellum (2017)
  • Author(s): Matsuda, K., Yoshida, M., Kawakami, K., Hibi, M., and Shimizu. T.
  • Journal Title: Scientific reports Volume: 7 Issue: 1 Pages: 11865-11865
  • DOI: 10.1038/s41598-017-10794-0
  • Peer Reviewed / Open Access
• [Journal Article] Medaka and zebrafish contactin1 mutants as a model for understanding neural circuits for motor coordination (2017)
  • Author(s): Takeuchi Miki、Inoue Chikako、Goshima Akiko、Nagao Yusuke、Shimizu Koichi、Miyamoto Hiroki、Shimizu Takashi、Hashimoto Hisashi、Yonemura Shigenobu、Kawahara Atsuo、Hirata Yutaka、Yoshida Masayuki、Hibi Masahiko
  • Journal Title: Genes to Cells Volume: 22 Issue: 8 Pages: 723-741
  • DOI: 10.1111/gtc.12509
  • NAID: 120006367981
  • Peer Reviewed / Open Access
• [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, Murakami Y.
  • Journal Title: Development, Growth & Differentiation Volume: 印刷中 Issue: 4 Pages: 228-243
  • DOI: 10.1111/dgd.12349
  • NAID: 120006406681
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Oncogenic role of rab escort protein 1 through EGFR and STAT3 pathway. (2017)
  • Author(s): Yun UJ, Sung JY, Park SY, Ye SK, Shim J, Lee JS, Hibi M, Bae YK, Kim YN.
  • Journal Title: Cell Death & Disease Volume: 8 Issue: 2 Pages: e2621-e2621
  • DOI: 10.1038/cddis.2017.50
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] REP1 inhibits FOXO3-mediated apoptosis to promote cancer cell survival. (2017)
  • Author(s): Song KH, Woo SR, Chung JY, Lee HJ, Oh SJ, Hong SO, Shim J, Kim YN, Rho SB, Hong SM, Cho H, Hibi M, Bae DJ, Kim SY, Kim MG, Kim TW, Bae YK.
  • Journal Title: Cell Death & Disease Volume: 8 Issue: 1 Pages: e2536-e2536
  • DOI: 10.1038/cddis.2016.462
  • Peer Reviewed / Int'l Joint Research
• [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] Osteocrin, a secretory peptide from heart, contributes to cranial osteogenesis in zebrafish (2016)
  • Author(s): Chiba A, Watanabe-Takano H, Terai K, Fukui H, Miyazaki T, Uemura M, Hashimoto H, Hibi M, Fukuhara S, Mochizuki N
  • Journal Title: Development Volume: 144 Pages: 334-344
  • DOI: 10.1242/dev.143354
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [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] PRMT8 as a phospholipase regulates Purkinje cell dendritic arborization and motor coordination. (2015)
  • Author(s): Kim, J.D., Park, K.E., Ishida, J., Kako, K., Hamada, J., Kani, S., Takeuchi, M., Namiki, K., Fukui, H., Fukuhara, S., Hibi, M,. Kobayashi, M., Kanaho, Y., Kasuya, Y., Mochizuki, N., Fukamizu, A.
  • Journal Title: Sci. Adv. Volume: 1 Issue: 11
  • DOI: 10.1126/sciadv.1500615
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Roles of cerebellar neural circuits in classical fear conditioning. (2017)
  • Author(s): Hibi M, Matsuda K, Yoshida M, Kawakami K, Shimizu T.
  • Organizer: International Workshop on Zebrafish Neural Circuits and Behavior.
  • Int'l Joint Research
• [Presentation] Formation and function of cerebellar neural circuits in zebrafish. (2017)
  • Author(s): Hibi M, Takeuchi M, Matsuda K, Ito T, Nimura T, Hara Y, Kuraku S, Kawakami K, Yoshida M, Shimizu T
  • Organizer: 第23回小型魚類研究会
  • Invited
• [Presentation] Tracing of zebrafish cerebellar neural circuits with a rabies virus system. (2017)
  • Author(s): Dohaku R, Osakada F, Hibi M.
  • Organizer: 第23回小型魚類研究会
• [Presentation] Tracing of zebrafish cerebellar neural circuits with a pseudotyped recmbinant rabies virus system. (2017)
  • Author(s): Dohaku R, Osakada F, Hibi M.
  • Organizer: 第40回日本神経科学大会
• [Presentation] 真骨魚類から見た小脳の進化と発生 (2016)
  • Author(s): 日比正彦、竹内未紀、松田光司、原雄一郎、榊原考将、種子島千春、山口信悟、吉田将之、工樂樹洋、清水貴史
  • Organizer: 日本進化学会第18回大会
  • Place of Presentation: 東京
  • Year and Date: 2016-08-25
  • Invited
• [Presentation] Role of Contactin1 in neural network formation for motor coordination in medaka and zebrafish. (2016)
  • Author(s): Takeuchi M, Goshima A, Inoue C, Shimizu K, Kawahara A, Yoshida M, Hibi M.
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: 横浜、神奈川
  • Year and Date: 2016-07-20
• [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、神奈川
  • Year and Date: 2016-07-20
• [Presentation] 真骨魚類に見る小脳の進化と発生 (2016)
  • Author(s): 日比正彦、竹内未紀、山口信悟、松田光司、原雄一郎、榊原考将、工樂樹洋、吉田将之、清水貴史
  • Organizer: 第121回日本解剖学会総会・全国学術集会
  • Place of Presentation: 郡山
  • Year and Date: 2016-03-28
  • Invited
• [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, 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] 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, 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] 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, 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, 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