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. 2021 Sep 28;9:733653. doi: 10.3389/fcell.2021.733653

FIGURE 8.

FIGURE 8

The sACspecific inhibitor TDI-10229 impairs [Ca2+]i responses to KCl in ejaculated human sperm. (A–D) Representative traces of [Ca2+]i responses to 50 mM KCl at (A,B) pH 6.8 and (C,D) pH 8.0 in ejaculated human sperm in non-capacitating conditions in (A,C) control and (B,D) after pre-incubation with TDI-10229. (E) Linear fit of the averaged normalized fluorescence in the first 20 s after KCl addition at pH 6.8 and pH 8.0. (F) Graph of slope values of the best-fit regression line for each condition. Mean values at pH 6.8 were as follows: control, 1.80 ± 0.82 amplitude units * s–1 (26 samples, 1836 sperm); and TDI-10229 incubation, 0.426 ± 0.276 amplitude units *s–1 (5 samples, 2279 sperm). Mean values at pH 8.0 were as follows: control, 1.73 ± 0.594 amplitude units * s–1 (26 samples, 3036 sperm); and TDI-10229 incubation, 0.491 ± 0.305 amplitude units *s–1 (5 samples, 2279 sperm). (G) Graph of the percentage of responsive sperm. Mean values at pH 6.8 were as follows: control, 66.94% ± 24.30 (26 samples, 1836 sperm); and TDI-10229 pre-incubation, 16.18% ± 10.10 (5 samples, 2279 sperm). Mean values at pH 8.0 were as follows: control, 77.04% ± 18.67 (26 samples, 3036 sperm); and TDI-10229 pre-incubation, 35.43% ± 15.77 (5 samples, 2279 sperm). *P < 0.050, **P < 0.010, and ***P < 0.001 by independent t-test. Error bars indicate standard deviation. Iono = 5 μM Ionomycin + 2 mM Ca2+.