Development of a single neural circuit visualization method using a novel neurotracer virus and a tissue transparency technique.
| Project/Area Number |
16K15059
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Integrative animal science
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
TANIGUCHI TAKAHIDE 東京農工大学, (連合)農学研究科(研究院), 准教授 (70282803)
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| Project Period (FY) |
2016-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | コロナウイルス / 神経回路 / コネクトーム解析 / イメージング / 発光タンパク質 / ウイルス / 蛍光タンパク質 / ニューロトレーシング |
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| Outline of Final Research Achievements |
Porcine hemagglutinating encephalomyelitis virus (PHEV) 67N strain efficiently infects rodent nerve cells. The virus strain infects adjacent neurons through synapses, and thus I conducted basic research to develop the 67 strain as a novel, safe neurotracer virus. We determined the whole genome sequence of 67N strain and found 20 amino acid substitutions in the S protein, which is important for the virus to infect neurons. Anti-N monoclonal antibodies (MAb) were produced using E. coli-expressed recombinant N protein as the antigen. I could detect PHEV-infected nerve cells and trace neuronal connection by the indirect fluorescent antibody method using these MAbs. And a highly sensitive and specific ELISA method for the detection of anti-PHEV antibodies was developed using recombinant N protein and extracted virus protein from PHEV infected cells as an antigen.
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| Academic Significance and Societal Importance of the Research Achievements |
PHEVはコロナウイルスとしては特異な神経細胞指向性を示すウイルスである。特に67N株は齧歯類の神経細胞に感染し、シナプスを介して隣接する神経細胞に感染が進み、病原性も低いことから安全性の高い新規ニューロトレーサーウイルスとして有望である。一方、最近インフルエンザ様呼吸器感染症を引き起こすPHEVが豚から分離され、遺伝子変異による組織指向性の急変の可能性が示唆された。本研究で得られた、67N及びOSN204株のゲノム塩基配列解析データやモノクローナル抗体、新規ELISA法はPHEVのニューロトレーサーとして開発のみならず、遺伝子変異による宿主や組織指向性の変化の機構解明に役立つものと思われる。
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Report
(5 results)
Research Products
(2 results)
