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. 2016 Jun 3;19(4):418–424. doi: 10.4103/1008-682X.181816

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Copyright: © The Author(s)(2017)

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.

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Expression of ClC-3 chloride channels in human spermatozoa. (a) and (b) the distribution of ClC-3 immunofluorescence (green) in spermatozoa from the normozoospermic and asthenozoospermic semen samples, respectively; scale bar = 10 μm. (c) the negative immunostaining control without the primary antibody in normal spermatozoa; the nuclei were stained with DAPI (blue); merge: merged images of the ClC-3 immunostaining and DAPI; scale bar = 10 μm. (d) ClC-3 immunofluorescence intensity in the neck and midpiece of spermatozoa (mean ± s.e., **P < 0.01, asthenozoospermia vs normozoospermia, n = 103 and 126, respectively). (e) ClC-3 expression analyzed by flow cytometry. The Y axes of histograms show cell numbers and the X axes show fluorescence intensity. Black peaks: control spermatozoa (without the primary antibody). Green and red peaks: spermatozoa from the asthenozoospermic (n = 26) and normozoospermic (n = 29) semen samples, respectively. Those spermatozoa with fluorescence intensity greater than the control cells were considered positive for ClC-3 expression. The expression levels of the ClC-3 proteins in spermatozoa from normozoospermic semen were higher than that in those from asthenozoospermic semen. DIC: differential interference contrast images; DAPI: 4’,6-diamidino-2-phenylindole.