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. 2018 Aug 6;8:11769. doi: 10.1038/s41598-018-30066-9

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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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Intracellular pH in human, bovine and ascidian spermatozoa exposed to metabolic enhancers. Intracellular pH (pH_i) in human (A), bovine (B) and ascidian (C) sperm divided in three groups treated with: (1) five metabolic enhancers (vitamins B6 and B12, 5 methyl THF, zinc and N-acetyl cysteine) (TRT group); (2) n-acetyl-cysteine (NAC group); and (3) medium alone (CNTRL group). Mean (±SE) values of pH_i in spermatozoa loaded with BCECF, arranged in TRT, NAC and CNTRL groups and then incubated for 0, 90 and 180 min. The fluorescence emission peaks (Fo) were measured at 535 nm after excitation at 440 and 490 nm. Statistically significant differences between groups for each evaluated time point are expressed as different capital (A vs. B; P < 0.01) letters.