Abberant α-Synuclein Confers Toxicity to Neurons in Part through Inhibition of Chaperone-Mediated Autophagy
Figure 6
Over-expression of human ASYN (WT, mutants) is toxic to rat cortical neurons.
(A) Five days-old cortical cultures were transduced with adenoviruses (MOI 150) expressing human ASYN (WT, ΔDQ/WT, A53T or ΔDQ/A53T) or EGFP (control virus) as described in Materials and Methods. 96 hrs later cells were lysed with a nuclear-sparing buffer and intact nuclei were counted in a hemacytometer. (B) Cortical cultures were treated as in A with slight modifications. 72 hrs post-infection, 3MA (10 mM) was added and intact nuclei were counted 24 hrs later. All data are presented as mean±SE of independent experiments and within each experiment triplicate samples per condition were assessed. (*p<0.05, **p<0.01, one way ANOVA followed by the Student-Newman-Keuls' test, comparing between cultures expressing various forms of ASYN and control EGFP; #p<0.05 comparing between cultures transduced with A53T and ΔDQ/A53T ASYN or between A53T ASYN transduced neurons+/−3MA).