Figure 5. Ttbk2 is critical for primary cilia stability on neurons.

(A–C) Representative images of cilia on indicated cell types throughout parts of the cerebellum and medulla. Sections from a mouse expressing an ARL13B-mCherry transgene (green) (Bangs et al., 2015) immunostained for γ-Tubulin to label centrosomes (white), and various cell specific markers such as Calbindin to label Purkinje cells (A), NeuN to label granule neurons (B), and FoxP2 to label neurons within the inferior olivary nucleus (C). Insets show boxed areas. Scale bar = 50 μm, 10 μm for insets. (D) Representative images illustrating cilia loss in the cerebellum 20 days after TMX treatment. Sections were immunostained for ARL13B to label cilia (red) and γ-Tubulin to label centrosomes (green). For quantification purposes images were taken at the nexus between the molecular layer (ML) and granule layer (GL) with the r PCL in the middle of the imaging field where there is an abundance of cilia. Scale bar = 50 μm. (E) Quantification of cilia loss after TMX treatment (n = 36 images counted, three animals, p<0.0001 student’s unpaired t-test, error bars indicate SEM).

Figure 5—figure supplement 1. Cilia are lost throughout the brain of Ttbk2^c.mut animals.

(A,B) Sagittal sections of Ttbk2^c.mut brains stained for AC3 to label cilia (red) and DAPI for nuclei. Neurons in the hippocampus (A) and the cortex (B) have lost cilia 3 months after tamoxifen injections in Ttbk2^c.mut animals. Scale bar = 20 μm.