|Budget Amount *help
¥3,400,000 (Direct Cost : ¥3,400,000)
Fiscal Year 2005 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 2004 : ¥2,600,000 (Direct Cost : ¥2,600,000)
We have been studying the mechanisms underlying cell death caused by alpha-synuclein using our inducible PC12 cell model of Parkinson's disease. We confirmed induction and suppression of alpha-synuclein in the absence and presence of doxycycline, respectively, with immunocytochemistry and western blotting experiments. We observed cytoplasmic inclusions stained for alpha-synuclein and ubiquitin in A53T mutant cell lines. In cell toxicity evaluation, we employed the trypan-blue exclusion assay and DAPI nuclear staining, finding increased apoptotic cell death after neuron-like differentiation and induction of A53T alpha-synuclein expression. Cell death we observed here was in an alpha-synuclein expression dependent manner.
We have proposed to elucidate the role for the NF-kappaB pathway in cell death caused by mutant alpha-synuclein. We found that cytoplasmic inclusions in A53T cell lines were positive for IkappaB immunostaining. We tried three NF-kappaB inhibitors including PDTC,DDTC, and SN5O, finding attenuation of A53T alpha-synuclein-caused cell death in the presence of the inhibitors.
Our findings suggest that the NF-kappaB pathway may play a role in mutant alpha-synuclein-caused cell death. In addition, the NF-kappaB pathway may be a potential target to treat neural cell death caused by alpha-synuclein.