Fig. 6: Ccdc57 deficiency decouples the BB polarity from the AO.
Quantification results in (c, e, g) are presented as mean ± SD plus sample dots. P-values were from unpaired two-tailed student’s t-test. *** P < 0.001. Arrows in micrographs indicate BB polarities (a, b, d) or AOs (f). a–c BBs in Ccdc57 −/− E1 cells lacked the rotational polarity. Ependymal tissues were immunostained for Cep164 to visualize BBs and γ-tubulin (a) or Centriolin (b) to visualize BFs. Diagrams (tilted side views) illustrate the protein localizations on the BB. Two neighboring BB patches, cropped from micrographs in Supplementary Fig. 4a and c, are presented for each mouse. Mean vector lengths (mean ± SD plus sample dots) (c) were measured from 20 BB patches (cells) for each P35 or P50 mouse. Unpaired two-tailed student’s t test. γ-tubulin: P = 2.5 × 10−30 for +/+ vs −/− (Mild), P = 1.1 × 10−53 for +/+ vs −/− (Severe); Centriolin: P = 6.4 × 10−12 for +/+ vs −/− (Mild), P = 2.6 × 10−25 for +/+ vs −/− (Severe). Theoretically, BB patches with full or no rotational polarity should have a mean vector length of 1 or 0, respectively. Mild, mild hydrocephalus; Severe, severe hydrocephalus. d, e Representative transmission EM micrographs (d) and quantification results (e). Each mean vector length (e) was calculated from a full-size EM micrograph, from mice ranging from P21 to P60. 100 full-size EM micrographs from six mice for each genotype. P = 2.0 × 10−64. f, g Ccdc57 deficiency did not impact the rotational polarity of axonemes. Representative EM images (f) were from the ependymal tissues of littermates. Quantification results (g) are presented as distributions of the mean angle of axonemes. 9 full-size EM micrographs from 3 mice ranging from P21 to P60 were quantified for each genotype. Each micrograph contained at least 6 clear axonemal cross-sections. h, i Decoupled BB polarities from AOs in Ccdc57 −/− E1 cells. Representative serial sections of 70 nm thickness (h) are presented. Illustrations depict the AO and the BB polarity of the cilia. One experiment (h) was performed. Radar chart (i) shows included angles between the two directions, measured respectively from 7 wild-type and 6 Ccdc57 −/− cilia. Source data are provided as a Source Data file.
