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. 2020 Jun 16;103(2):343–356. doi: 10.1093/biolre/ioaa093

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© The Author(s) 2020. Published by Oxford University Press on behalf of Society for the Study of Reproduction.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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(A) Dose–response curve for the ouabain inhibition of NKA from rat sperm. NKA activity was measured under saturated conditions of Na⁺, K⁺, and Mg²⁺ and at the indicated ouabain concentrations. Curves represent the best fit of the experimental data and indicate the presence of two enzyme components, one highly sensitive (NKAα4) and the other one resistant (NKAα1) to ouabain. Each value is the mean, and error bars represent the SEM of three experiments performed in triplicate. (B) Measurement of sperm total motility using computer-assisted sperm analysis (CASA) on rat epididymal spermatozoa. Cells in Whittens medium were treated for the indicated times with no ouabain (control), 1 nM ouabain (NKAα4 inhibition), or 1 μM ouabain (NKAα4 plus NKAα1 inhibition). Sperm motility was measured under non-capacitating conditions. Values are mean ± SEM [66, 77].