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. 2019 Nov 1;9:15864. doi: 10.1038/s41598-019-52436-7

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© The Author(s) 2019

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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DNAH6-mutant patients have abnormal, disorganized sperm axonemes. (a,b) Spermatozoa from normal control (NC) and DNAH6-mutant patients (P1 and P3) were double-labeled with antibodies against acetylated α-tubulin to identify axonemes and basal bodies (green), DNAH1 as a marker of the inner dynein arm (red), and SPAG6 as marker of CPC (red). Both α-tubulin and DNAH1 or SPAG6 were colocalized along the full length of axonemes in sperm flagellum from the NC (upper panel). In contrast, in MMAF patients (P1 and P3) (lower panels), DNAH1 or SPAG6 expression was markedly reduced in the axonemes compared to NC. (c,d) Spermatozoa from control and DNAH6-mutant patients (P1 and P3) was double-labeled with antibodies against acetylated α-tubulin to identify axonemes and basal bodies (green), RSPH1 as a marker of the radial spoke (red), and AKAP4 as a marker of the fibrous sheath (red). Scale bars = 5 μm.