2015 Fiscal Year Final Research Report
Establishment of cellular and rodent models for amyotrophic lateral sclerosis using recombinant viral vectors
Project/Area Number |
24500428
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Nerve anatomy/Neuropathology
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Research Institution | Tokyo Metropolitan Institute of Medical Science |
Principal Investigator |
WATABE Kazuhiko 公益財団法人東京都医学総合研究所, 運動・感覚システム研究分野, 副参事研究員 (30240477)
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Project Period (FY) |
2012-04-01 – 2016-03-31
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Keywords | 運動ニューロン / TDP-43 / FUS / 筋萎縮性側索硬化症 / 凝集体 / 組換えウイルス / プロテアソーム / オートファジー |
Outline of Final Research Achievements |
Formation of TDP-43- or FUS-positive cytoplasmic aggregates in neuronal and glial cells is one of the pathological hallmarks of amyotrophic lateral sclerosis (ALS). We have demonstrated that inhibition of protein degradation pathways enhanced adenovirus-induced neuronal cytoplasmic aggregate formation of TDP-43 and FUS in vitro and in vivo. We then produced recombinant adeno-associated virus type 9 (AAV9) and lentivirus vectors encoding wild type and mutant TDP-43 or FUS, and those encoding shRNAs for protein degradation machineries and demonstrated their long-term retrograde transduction of the foreign genes and formation of cytoplasmic aggregates in adult mouse facial and spinal motoneurons. We also performed time-lapse imaging analysis of neuronal and glial cells infected with adenoviruses encoding TDP-43 and FUS cDNAs under conditions of proteasome inhibition to examine cytoplasmic aggregate formation, cell death, and cell to cell spreading of these aggregates.
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Free Research Field |
神経科学・神経解剖学・神経病理学
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