Fig. 5. Mitochondrial nitrative damage within hyperactive h-αS–loaded neurons.
(A to D) iR26-αS mice were injected intravagally with a solution containing Cre- and hM3Dfl-AAVs, treated with either saline (n = 5) or CNO (n = 5) during weeks 4 and 5, and sacrificed at 5 weeks after AAV injection. To detect nitrated NDUFB8 (A and B) or nitrated SOD2 (C and D), sections of the MO were processed for either 3-NT/NDUFB8 or 3-NT/SOD2 PLA. They were also labeled with anti–h-αS. Representative confocal images show fluorescent PLA dots within h-αS–immunoreactive DMnX neurons. Scale bars, 20 μm (A and C). Fluorescent intensity and number of PLA dots were measured within h-αS–positive DMnX neurons. Approximately 15 neurons per animal were analyzed, and for each animal, intensity values and PLA counts were averaged. Data were then calculated as percentage of the mean value in the saline group (B and D). Box and whisker plots show median, upper and lower quartiles, and maximum and minimum as whiskers. *P < 0.05 and **P < 0.01.