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. 2018 Jun 15;8:9202. doi: 10.1038/s41598-018-27379-0

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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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Bright field and fluorescence microscopy of cauda epididymal sperm from WT and Dcaf17^−/− adult mice. Representative images of Diff-Quick staining (bright field images, top panels) and MitoTracker staining (fluorescence images, bottom panels) of epididymal sperm spreads from WT (A,C) and Dcaf17^−/− (B,D) adult mice. The WT sperm (A,C) show typical hook-shaped head, patent midpiece and tail morphology. Whereas, the Dcaf17^−/− sperm (B,D) show variety of morphological defects in head shape, midpiece and tail. To categorize different sperm defects in Dcaf17 KO mice we analyzed total 150 sperms from 3 different mice. Major categories of sperm head defects were triangular (cupcake) (B1,D1,D3), oval (B5,D2,D4,D5) or amorphous (B2,B3) shaped with many times either bent head or coiled midpiece surrounding the head. Fluorescently labeled WT sperm (C) shows normal crescent shaped acrosome (green), uniformly distributed mitochondria (red) along the midpiece and highly condensed nucleus (blue). Dcaf17^−/− sperm (D1–5) show spatially dysmorphic sperm structure with abnormal acrosome (green), ectopic localization of mitochondria (red) and diffused chromatin structure (blue). Images C and D1–5 are merged fluorescence images of sperm stained for acrosome (green), midpiece (red) and nucleus (blue). Separate fluorescence images for each sperm acrosome, midpiece and nucleus staining are shown in supplementary figure S5. Magnification of bright images (A,B1–5) is 400X and magnification of fluorescence images (C,D1–5) is 1000X. Scale bar in the image A is 1 mm and in the image C is 10 µm.