Fig. 6

VPS35 overexpression fails to protect against nigrostriatal dopaminergic neurodegeneration induced by human WT-α-Syn in adult rats. A-B) Representative photomicrographs of (A) human VPS35 (V5) and (B) human α-synuclein (Syn211) immunostaining in the substantia nigra (SNpc) and striatum of adult rats at 14 weeks after co-injection of AAV2/6 vectors expressing WT-α-Syn together with VPS35 (WT or D620N) or empty control (MCS). Non-injected (left) and injected (right) hemispheres are indicated. C) Fluorescent immunostaining indicating marked co-localization of human VPS35 (WT or D620N) and human WT-α-Syn with TH-positive dopaminergic neurons of the injected rat substantia nigra at 14 weeks. D) Representative photomicrographs of anti-TH immunostaining in the rat substantia nigra (SNpc) at 14 weeks after the co-injection of AAV2/6 vectors expressing combinations of human VPS35 (WT or D620N), human WT-αSyn, or empty control (MCS). E) Unbiased stereological quantification of TH-positive dopaminergic and total Nissl-positive neurons in the substantia nigra at 14 weeks post-injection of AAV vectors. Data are expressed as percent cell loss relative to non-injected nigra with bars indicating the mean ± SEM (n = 7–8 animals/group). *P < 0.05 or **P < 0.01 by one-way ANOVA with Tukey’s multiple comparison test, as indicated. n.s., non-significant. F) Representative photomicrographs of immunostaining for TH-positive nerve terminals in the striatum at 14 weeks after AAV vector delivery in injected and non-injected hemispheres. G) Quantitation of optical density of TH-positive immunostaining in the striatum. Data are expressed as percent loss of TH-positive fibers relative to the non-injected side, with bars indicating the mean ± SEM (n = 7–8 animals/group). n.s., non-significant by one-way ANOVA with Tukey’s multiple comparison test