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. 2016 Aug 18;6:31816. doi: 10.1038/srep31816

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Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Testicular, epididymal and vas deferens spermatozoa immunostained with antibodies to centrin 20H5 (green, arrowheads), AK15 γ-tubulin (red, arrows) and DNA (blue). (A) In testicular sperm, both the proximal and distal centrioles at the implantation fossa are detected by centrin (green, arrowheads) with the majority of AK15 γ-tubulin observed in the cytoplasmic droplet attached to the upper sperm axoneme (A: red, arrow; inset: DIC) or surrounding the centriole pairs. (B) Caput epididymal spermatozoa demonstrate strong proximal and distal centriole detection with centrin 20H5 (green, arrowheads), while remaining γ-tubulin coalesces adjacent to the centriole pairs at the sperm axoneme’s basal body region (red, arrows). (C) In the upper corpus, a majority of the spermatozoa begin to show a reduction in centrin intensity at the implantation fossa (green, arrowheads), but not γ-tubulin (red, arrows). (D) In the middle corpus, spermatozoa can retain a strong γ-tubulin signal (upper sperm: red, arrow) and the centrin intensity in the distal centriole is frequently lost, as the proximal centriole continues to further diminish (green, arrowhead). Some spermatozoa lose both centrin and γ-tubulin detection at this stage (lower sperm: arrow). (E–G) In spermatozoa isolated from the lower corpus (E) cauda (F) and vas deferens (G) centrin no longer detects any centrioles (green) even though most sperm retain detectable γ-tubulin at the implantation fossa (red, arrows). All images are triple-labeled for centrin 20H5 (green), AK15 γ-tubulin (red) and DNA (blue). Insets: DIC: differential interference contrast. Bars: μm.