| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| 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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