Alpha-synucleinopathy in dopaminergic neurons after exposure to 6OHDA, MPP⁺ or rotenone.

(A). Upper panel: Representative picture of aggregated α-syn (in green) inside TH-expressing neurons (in red) (white arrows) after double-immunofluorescence microscopy, obtained with an antibody anti-aggregated α-syn (scale bar 100 μm). Lower panel: Representative picture of phase contrast microscopy reveals intracellular Lewy body-like structures (scale bar 100 μm). (B, C). Representative picture of α-syn, obtained with an antibody anti-α-syn and the area of α-syn staining overlapping with TH staining, corrected by the number of TH neurons, after an injury with 6OHDA (48 h), MPP+ (48 h) or rotenone (24 h). A value at 1 is equivalent to 7,1 +/- 0,38 neurons. (D). Representative pictures of LC3 staining (red) in TH expressing neurons (green) and the segmentation resulting from data processing (white). Arrow: TH neurons; star: non TH neuron showing LC3 staining. Data processing removed background signal to consider only dot structures in TH expressing neurons. (E). LC3 staining in dopaminergic neurons in presence of the toxins. A value at 1 is equivalent to 1.7 +/- 0,2 neurons. All values were expressed as mean +/- SEM; n = 6/group; *, p<0,05 with One-way ANOVA followed by Dunnett’s test.