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. 2020 Mar 24;10:5335. doi: 10.1038/s41598-020-62091-y

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© The Author(s) 2020

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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Normal testis and epididymal sperm localization and relocation after acrosomal reaction of IZUMO1 in Spaca6 KO context. (a) Immunofluorescence staining of IZUMO1 and DAPI staining of permeabilized testicular sections from WT and Spaca6 KO C57BL/6 mice. The labeling of IZUMO1 (green) was carried out using a rat monoclonal anti-mouse IZUMO1 primary antibody, recognized by a secondary anti-rat goat antibody coupled to Alexa Fluor 488. The nuclei of the testicular cells were labeled with DAPI at 10 μg/mL. IZUMO1 was similarly detected in round and elongated spermatids (RS and ES) from WT and Spaca6 KO testes. A control condition with an incubation of the tissue with the secondary antibody alone was performed to check the specificity of the antibodies and to evaluate the background noise (negative control). Sg: Spermatogonia, Sc: Spermatocyte, RS, Round spermatid, ER: Elongated Spermatid, SC: Sertoli Cell. (b) Immunofluorescence staining of IZUMO1 and DAPI and PSA labeling of epididymal sperm from WT and Spaca6 KO males before or after acrosome reaction. While IZUMO1 is essentially localized in acrosomal cap in acrosome-intact sperm, it is more distributed on the sperm head after acrosome reaction. The absence of SPACA6 has no effect on IZUMO1 sperm localization and relocation after acrosome reaction.