title: Macroautophagy and the proteasome are differently involved in the degradation of alpha-synuclein wild type and mutated A30P in an in vitro inducible model (PC12/TetOn).
abstract: Many data suggest that alpha synuclein (α-syn) aggregation is involved in Parkinson's disease (PD) neurotoxicity and is accelerated by the pathogenetic point mutation A30P. The triplication of α-syn gene has been linked to early-onset familial PD, suggesting that the cellular dosage of α-syn is an important modulator of its toxicity. To verify this point, we developed an inducible model of α-syn expression (both wild type [WT] and mutated A30P) in rat PC12/TetOn cells. At low expression level, both α-syn(WT) and (A30P) did not aggregate, were not toxic, and displayed a protective action against oxidative stress triggered by hydrogen peroxide (H(2)O(2)). By increasing α-syn expression, its antioxidant function was no longer detectable as for the A30P form, but again no aggregation and cell death were present both for the WT and the mutated protein. To clarify why α-syn did not accumulate at high expression level, we inhibited macroautophagy by 3-methyladenine (3-MA) and the proteasome by MG132. In presence of 3-MA, α-syn(WT) accumulated, A11 anti-oligomer antibody-positive aggregates were detectable, and cell toxicity was evident, while proteasome inhibition did not increase α-syn(WT) accumulation. Macroautophagy or proteasome inhibition slightly increased α-syn(A30P) toxicity, with no detectable aggregation. This model can provide useful details about α-syn function, aggregation, and degradation pathways.
authors: Department of Neuroscience, Mario Negri Institute for Pharmacological Research, Milan, Italy.
authors: S
authors: S
authors: Batelli
authors: E
authors: E
authors: Peverelli
authors: S
authors: S
authors: Rodilossi
authors: G
authors: G
authors: Forloni
authors: D
authors: D
authors: Albani
mesh_terms: D000818:Animals; D001343:Autophagy / Q000235:genetics*; D015153:Blotting, Western; D002470:Cell Survival / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D018384:Oxidative Stress / Q000235:genetics; D016716:PC12 Cells; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D051381:Rats; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein; D046988:Proteasome Endopeptidase Complex