Fig. 5: Defective ciliation of ependyma in trip6−/− mice.
Deconvolved immunofluorescence microscopy images of the ependyma of three trip6−/− and two control trip6+/− mice (all at P4) depict, in high spatial resolution, adhesions (labeled by an anti-β-catenin antibody), cilia (anti-acetylated α-tubulin labeling), and basal bodies (anti-γ-tubulin labeling). An animated 3-D reconstruction of the images is presented in Supplementary Movies 3, 4, and 5. There are no obvious adhesion defects in the ependyma, neither in the LV (a cf. b, c, f) nor in the 3rd ventricle (d cf. e), as demonstrated by comparison of control (a, d) with trip6−/− (b, c, e, f) specimens (β-CATENIN). In contrast, in the LV, defective ciliogenesis in trip6−/− mice of mild (b) or severe (c) phenotype (degree of cilia absence) is demonstrated by ependyma areas presenting with a thin layer of acetylated α-tubulin labeling (Ac.TUB, arrows). For comparison, normal cilia bundles are indicated by arrowheads, in control (a) and trip6−/− (b, c) specimens. (see also Fig. 7dcf. e, for higher resolution images of cilia). Regular formation and ventricular localization of basal bodies, in trip6−/− ependymal cells (e, f) of 3rd and LVs, is visualized by the punctate γ-tubulin staining pattern. The brackets indicate clusters of basal bodies within single ependymal cells (γ-TUB; d–f). Scale bars: 10 μm.
