Figure 11.

αS species capable of inducting intracellular αS aggregates can be passaged from cell lysates. (A) Astrocyte cultures with rAAV2/1-mediated overexpression of human wild-type αS treated with cell lysate from astrocyte cultures containing αS aggregates via overexpression of human wild-type αS and addition of human A53T- derived αS fibrils. At five days post-seeding, this resulted in pSer129 (green) aggregates in both the cell body and processes, which overlapped with SNL-4 (red). This was compared to astrocyte cultures with rAAV2/1-mediated overexpression of human wild-type αS and treated with cell lysates from astrocyte cultures treated with human A53T-derived αS fibrils (B), rAAV2/1-mediated overexpression of human wild-type αS (C), or untreated cultures (D). In the latter 3 conditions, at 5 days post-seeding there was uniformly distributed overexpression of αS in the absence of hyperphosphorylated αS aggregates (B,C,D). Cultures were counterstained with DAPI (blue). Bar scale = 100 μm; insets = 25 μm.