Fig 1.

Lentiviral-mediated overexpression of α-syn. (A) Western blot analysis shows the overexpression of normal and mutated (A53T and A30P) human and rat α-syn in the SH-SY5Y human neuroblastoma cell line. All α-syn forms are expressed at similar levels for the same amount of viral particles. Protein (25 μg per lane) were loaded for the noninfected cells (NI) and cells transduced with lentiviral vectors encoding for cytoplasmic LacZ, rat α-syn, wild-type (HWT), and mutated forms of human α-syn. The 19-kDa α-syn bands (α-syn) were detected with a polyclonal rabbit Ab generated against the 101- to 124-aa sequence of human α-syn. This Ab recognizes both human and rat α-syn on Western blot. The amount of protein loaded was checked by reprobing the same membrane with an α-tubulin Ab (α-tub). (B–D) Lentiviral vectors encoding for wild-type and mutated human α-syn were stereotactically injected in the substantia nigra of rats. The nigral dopaminergic neurons were specifically labeled with a TH Ab (B). Detection with an α-syn polyclonal Ab revealed a significant overexpression of A30P α-syn (C) in the injected hemisphere. No α-syn staining was observed on the contralateral side. Double staining (D, yellow-orange color) shows a large proportion of TH-IR neurons overexpressing α-syn. (Scale bars = 200 μm.)