Fig. 4

Effects of cyclosporin A, a CypD inhibitor on sperm motility, phosphotyrosine proteins, mPTP opening, p-GSK3α(Ser21), MMP, and ATP production. A Effects of cyclosporin A (CsA) on sperm motility. Total and progressive motility of spermatozoa were significantly increased under capacitation condition. CsA significantly increased total and progressive motility of spermatozoa (n = 4). B Representative western blot images of the effects of CsA on phosphotyrosine (p-Tyr) proteins of spermatozoa. CsA markedly increased p-Tyr proteins of spermatozoa. At least three independent experiments were subjected to western blot analysis. C Representative images of calcein fluorescence in CsA-treated spermatozoa. CsA markedly increased calcein fluorescence intensity in the sperm midpiece. Bar, 5 μm. D Effect of CsA on mPTP opening. CsA significantly increased calcein fluorescence, indicative of decreased mPTP opening in spermatozoa under capacitation condition (n = 4). E Representative images of TMRE fluorescence in CsA-treated spermatozoa. TMRE fluorescence intensity was markedly increased in the sperm midpiece under capacitation and was further potentiated by CsA. Bar, 5 μm. F Effects of CsA on sperm mitochondrial membrane potential (MMP). MMP of spermatozoa was significantly increased under capacitation condition. CsA significantly increased the MMP of spermatozoa (n = 4). G Effect of CsA on p-GSK3α(Ser21). No change was observed in p-GSK3α(Ser21) level by CsA treatment in spermatozoa under capacitation condition. At least three independent experiments were subjected to western blot analysis. H Effect of CsA on ATP production in spermatozoa. CsA significantly increased ATP production in spermatozoa (n = 4). Data are presented as mean ± SD. Significant differences on one-way ANOVA (p < 0.05) are indicated by different letters. *p < 0.05 and ***p < 0.001 indicates significant difference from control by Student’s t-test. AU, arbitrary units