Fig. 6.

Loss of PINK1 decreases ciliary signaling in mouse striatum in vivo. (A and B) Confocal images of sections of dorsal striatum from 5-mo-old WT, LRRK2^RC, PINK1^KO, and LRRK2^RC/PINK1^KO mouse brains. (A) Cholinergic interneurons were labeled using anti-choline acetyltransferase (ChAT) antibody (green), primary cilia were labeled using anti-AC3 antibody (magenta, white arrow), and nuclei were stained with DAPI (blue). (B) Astrocytes were labeled using anti-glial fibrillary acidic protein antibody (green), primary cilia were labeled using anti-ADP ribosylation factor like GTPase 13B antibody (magenta, white arrow) and nuclei were labeled using DAPI (blue). (C) Quantitation of the percentage of ChAT⁺ neurons containing a cilium and (D) their cilia length. Similar analyses for astrocytes are shown in (E and F). (G) Confocal images to identify ChAT⁺ neurons and their cilia as in A (Left columns) coupled with RNAscope in situ hybridization to detect GDNF transcripts (Right columns), segregated by ciliation status in WT, LRRK2^RC, PINK1^KO, and LRRK2^RC/PINK1^KO mouse brains as indicated. (H) Quantitation of GDNF RNA dots per neuron or (I) segregated as a function of ciliation status. Error bars represent SEM from N = 3, 4 mouse brains, with >30 ChAT⁺ neurons and 25 astrocytes scored per brain. In bar charts, the black circle represents LRRK2^WT while red squares LRRK2^RC mice. Ordinary two-way ANOVA with Sidak’s multiple comparison test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (Scale bar, 10 µm.)