Figure 1

αS seeded aggregate formation in rAAV2/1-mediated αS expression of mixed neuronal-glial cultures. Double immunofluorescence with SNL-4 (red) and pSer129 (green): (A) In untreated cultures, αS demonstrated predominantly a punctate staining profile reflecting a presynaptic distribution, as previously demonstrated [39, 40] and (B) rAAV2/1-mediated overexpression of αS increases both the presynaptic and nuclear staining intensity for αS. In these cultures, pSer129 demonstrates a neuritic staining profile that does not appear to be due to αS. (C) Addition of fibrils alone resulted in pSer129 staining of processes also seen in blank controls, but not overlapping with SNL-4. (D) Both rAAV2/1-mediated overexpression of αS and the addition of fibrils resulted in pSer129 immunoreactive inclusions in both the cell body and processes that overlaps with SNL-4. In cultures derived from SNCA null mice, both (E) untreated and (F) addition of fibrils resulted in a neuritic staining profile with pSer129, in the absence of SNL-4 staining. The addition of fibrils did not increase the amount of pSer129 neuritic staining. Cultures were counter stained with DAPI (blue). Bar scale = 100 μm; insets = 25 μm.