Fig. 8

VPS35 overexpression does not attenuate human WT-α-Syn-dependent pathology in the rat substantia nigra. A-P) Immunohistochemical staining of rat substantia nigra pars compacta (SNpc) at 14 weeks following intranigral co-injection of AAV vectors expressing human WT-α-Syn with human VPS35 (WT or D620N) or empty control (MCS). Immunoreactivity for (A-D) pS129-α-Syn, (E-H) astrocytes (GFAP-positive), (I-L) total microglia (Iba1-positive), and (M-P) activated microglia (CD68-positive), in the injected SNpc are shown. The non-injected SNpc from the α-Syn/MCS control group is shown for comparison. Q-U) Quantitation of each pathological marker using HALO analysis software. SNpc sections with pS129-α-Syn, GFAP, Iba1 (total or activated) and CD68 immunostaining were quantified as the percent area occupied by each marker in the ipsilateral (injected) or contralateral (non-injected) hemispheres. Activated Iba1-positive microglia were identified from total microglia based on morphological criteria using Halo (T). Bars represent mean ± SEM (n = 5–8 animals/group). *P < 0.05 or **P < 0.01 by one-way ANOVA with Tukey’s multiple comparison’s test, as indicated