Figure 3: Intranigral infusion of acrolein induced protein conjugation and α-synuclein aggregation in the nigrostriatal dopaminergic system of rat brain.

(A and C): For a dose-dependent effect, ARC (15–150 nmoles) was locally infused in the substantia nigra (SN) and rats were sacrificed 7 days after intranigral infusion of acrolein. (B and D): For a time-dependent effect, acrolein (150 nmoles) was locally infused in the SN and rats were sacrificed at the time indicated. Western blot assay was employed to detect FDP-lysine (A,B) and α-synuclein (C,D) in the SN. Each lane contained 25 μg proteins for all experiments. Graphs show statistical results from relative optical density of bands on the blots estimated by Image J. Values are the mean ± S.E.M. (n = 3/group). *p < 0.05 in the acrolein-infused SN compared with the intact SN by one-way analysis of variance (one-way ANOVA) and followed by the LSD test as a post-hoc analysis. (E) Representative confocal microscopic data showed co-localization of FDP-lysine and α-synuclein immunoreactivities in the infused SN of rat brain. Acrolein (150 nmoles) was locally infused in the SN and rats were sacrificed 48 hrs after intranigral infusion of acrolein. Scale bars: 10 μm. Results were repeated in independent experiments.