Fig. 3: Ablation of Centlein leads to acephalic spermatozoa.
From: The missing linker between SUN5 and PMFBP1 in sperm head-tail coupling apparatus
a Fewer sperm heads are present in Centlein−/− epididymis. The H&E staining of the caudal epididymis of Centlein+/+, Centlein+/−, and Centlein−/− mice are shown. The spermatozoa within the Centlein-deficient caudal epididymis appear to be stained less with hematoxylin compared with those in the Centlein+/+ and Centlein+/− caudal epididymis. Biologically independent mice for each genotype were examined in three separate experiments with similar results. b The sperm counts in the caudal epididymis of Centlein−/− mice was significantly reduced compared with Centlein+/+ and Centlein+/− mice (n = 6 independent experiments). Data are presented as mean ± SEM. A two-tailed Student’s t test was performed, ***P  < 0.001, ****P  0.0001. c The Centlein-null spermatozoa are headless. Single-sperm PNA (green) staining was performed using Centlein+/+, Centlein+/−, and Centlein−/− spermatozoa. Nuclei were stained with DAPI (blue). d Proportion of decapitated tails in Centlein+/+, Centlein+/−, and Centlein−/− caudal epididymis (n = 6 independent experiments). Data are presented as mean ± SEM. A two-tailed Student’s t test was performed, ****P < 0.0001. e Ultrastructure of Centlein+/+ and Centlein−/− spermatozoa from caudal epididymis showing that the Centlein-null spermatozoa had no sperm head and contained a residual droplet of cytoplasm at the top of the flagellum with misarranged mitochondria inside. Nu: nuclear, M (red): mitochondrion, AX (blue): axoneme. The red asterisk indicates the missing microtubule doublets of axoneme in Centlein-null spermatozoa. Biologically independent mice were examined in three separate experiments with similar results. f Ultrastructure of the midpiece, principal piece, and end piece of Centlein+/+ and Centlein−/− spermatozoa from caudal epididymis. M (red): mitochondrion, AX (blue): axoneme, Od (yellow): outer dense fibers. The red asterisk indicates the missing microtubule doublets of axoneme in Centlein-null spermatozoa. g–i Quantification ratio of abnormal axoneme in midpiece (g), principal piece (h), and the end piece (i) of Centlein+/+ and Centlein−/− spermatozoa (n = 3 independent experiments). Data are presented as mean ± SEM. A two-tailed Student’s t test was performed, **P < 0.01. Source data are provided as a Source Data file. Blue dots indicate Centlein+/+mice, green dots indicate Centlein+/− mice, and red dots indicate Centlein−/− mice.
