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. 2021 Aug 13;12:4926. doi: 10.1038/s41467-021-25227-w

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a The proportion of decapitated tails in Centlein^+/+, Centlein^+/−, and Centlein^−/− corpus and caput epididymis (n = 6 independent experiments). Data are presented as mean ± SEM. A two-tailed Student’s t test was performed, ****P < 0.0001. b PAS and hematoxylin staining were performed in Centlein^+/+ and Centlein^−/− mouse. The mature sperm head could still be detected at stages IX–X in Centlein-deficient testes. The arrows indicate the orientation of the sperm heads. L: lumen, BM: basement membrane, P (red): pachytene spermatocyte, D (purple): diplonema spermatocyte, rST (green): round spermatid, eST (black): elongating spermatid, M (orange): meiotic spermatocyte, spz (blue): spermatozoa. Biologically independent mice were examined in three separate experiments with similar results. c The proportion of different stages of tubule cross-sections in Centlein^+/+ and Centlein^−/− mice (n = 3 independent experiments). d Centlein-null spermatids have lost their orientation toward the basement membrane during spermiation in stage V–VIII seminiferous epithelia. The arrows indicate the orientation of the sperm heads. L: lumen, BM: basement membrane. e Quantification ratio of spermatids with abnormal orientation in Centlein^+/+ and Centlein^−/− testes (n = 3 independent experiments). Data are presented as mean ± SEM. A two-tailed Student’s t test was performed, **P < 0.01. f The coupling apparatus could not be tightly attached to the sperm head in Centlein-null mice. TEM analyses of the stepwise development of the coupling apparatus were performed in Centlein^+/+ and Centlein^−/− testes. The red asterisk indicates destroyed coupling apparatus. The yellow asterisk indicates the gap between the nuclear envelope and coupling apparatus. Nu: nuclear, Ac: acrosome, Bp (blue): basal plate, Cp (orange): capitulum, Sc (red): segmented column, Pc (white): proximal centriole, Dc (purple): distal centriole, An (yellow): annulus, Od (green): outer dense fibers. Biologically independent mice were examined in three separate experiments with similar results. 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.