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. 2021 Feb 22;22(4):2187. doi: 10.3390/ijms22042187

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Localization of SUN5 and immunostaining in human spermatozoa from control and mutated subjects. (A) Localization of the SUN5 protein at the base of the sperm head, where it functions to anchor the sperm head to the flagellum. (B,C) Sperm cells from mutated subjects P0166 and P0177 (B) and P0504 (C) in addition to those from fertile controls were stained with anti-SUN5 (1:200, green) and anti-acetylated tubulin (1:2000, red) antibodies. DNA was counterstained with 4′,6-diamidino-2-phenylindole (DAPI), showing sperm nuclear DNA (blue). The SUN5 immunostaining (green) marked a punctiform signal at the head–tail junction. The co-staining with alpha-tubulin, specific for the flagella, confirm the localization. In contrast, the SUN5 signal is absent from P0166, P0177, and P0504, confirming the deleterious effect of the mutation. KASH: Klarsicht, ANC-1, Syne Homology; LINC: linker of nucleoskeleton and cytoskeleton.