Fig. 8. Administration of LRRK2 kinase inhibitor ameliorates neuropathology and motor behavioral deficits in a mouse model of synucleinopathy.

Non-transgenic and α-syn-transgenic mice were injected with vehicle or LRRK2 kinase inhibitor HG-10-102-01 (10 mg/kg) five times weekly for 4 weeks. (A) Representative images from immunohistochemistry analysis for astrogliosis (GFAP) and microgliosis (IBA-1) in the neocortex, striatum, and hippocampus of injected mice. Data are means ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001 (One-way ANOVA with Tukey’s multiple comparison post-hoc test). n = 6 per group. Scale bar, 25 μm. (B to J) Quantitative gene expression analysis from brains of injected mice. Expression of Tnfα (B), Il-6 (C), Il-1β (D), Il-10 (E), Cxcl1 (F), Ccl2 (G), Ccl4 (H), Ccl5 (I), and Egr2 (J) was determined by quantitative PCR. Data are means ± SEM. *P < 0.05, **P < 0.01 and ***P < 0.001 (One-way ANOVA and Tukey’s multiple comparison post-hoc test) n = 4 per group. (K to P) Behavioral analysis of injected mice. (K and L) Motor learning and function were determined by rotarod. The rotarod time (K) and distance (L) were analyzed. *P < 0.05 (One-way ANOVA with Tukey’s multiple comparison post-hoc test). n = 5 per group. (M) Neuromuscular impairment was determined by wire hang test. Data are means ± SEM. n = 5 per group. (N to P) Total activity (N), ambulatory time (O), and total distance (P) were analyzed by open field test. Data are means ± SEM. *P < 0.05 (One-way ANOVA with Tukey’s multiple comparison post-hoc test). n = 5 per group. (Q) Western blot analysis of whole mice brain lysates. The blot was probed with phospho-S54 NFATc2, total NFATc2, and β-Actin. NFATc2 band intensity was determined by densitometric quantification and normalized to β-Actin. ***P < 0.001 (One-way ANOVA and Tukey’s multiple comparison post-hoc test). n = 5 per group.