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. 2018 Dec 3;8:17531. doi: 10.1038/s41598-018-35719-3

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© The Author(s) 2018

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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Effect of the lack of CRISP1 and CRISP4 on in vitro sperm fertilizing ability: (A–D) Capacitated sperm from WT or DKO Group 1 (DKO1) or Group 2 (DKO2) males were co-incubated with COC for 3 h (A,B), ZP-intact eggs for 3 h (C) or ZP-free eggs for 1 h (D). At the end of all incubations, fertilization was evaluated by the presence of decondensing sperm heads within the egg cytoplasm. In (B) WT* corresponds to WT capacitated sperm treated to reach the low viability rates of Group 2 DKO sperm and DKO2* represents capacitated sperm from Group 2 DKO mice subjected to a swim up procedure to select viable sperm. Data are mean ± SEM, n = 4. Different letters indicate significant differences between groups (a vs b, p < 0.001). (E) Hoescht-stained capacitated sperm were co-incubated with COC for 15 min and the number of sperm within the cumulus matrix was determined. Data are mean ± SEM, n = 4, *p < 0.05. (F) Capacitated sperm were co-incubated with ZP-intact eggs for 30 min and the number of sperm bound to the ZP was evaluated. Data are mean ± SEM, n = 4, *p < 0.05.