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

2017 Fiscal Year Final Research Report

• 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
• 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

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.

Free Research Field

発生生物学