FIGURE 1.

Impacts of synthetic electrophiles on the generation of mitochondrial ROS, motility, and vitality of human spermatozoa. Left panels depict flow cytometry analyses, revealing the dose-dependent impact of electrophiles on the generation of mitochondrial ROS by human spermatozoa. Mitochondrial ROS in live cells was measured with MSR in conjunction with SYTOX Green following a 2-h exposure. Pos, positive control incubations involving exposure to arachidonic acid (5 μm) for 5 min at 37 °C. Right panels demonstrate the impact of the same electrophiles on sperm motility (filled bars) and vitality as assessed by SYTOX Green (open bars). The electrophiles examined comprised benzoquinone (A), iodoacetamide (B), NEM (C), ethyl vinyl ketone (D), and acrylamide (E). All of the electrophiles tested except acrylamide demonstrated a capacity to stimulate mitochondrial ROS generation and suppress motility in live cells. Data were analyzed by ANOVA, and values are presented as means ± S.E. (error bars); ***, p < 0.001; **, p < 0.01 for differences with negative control by Fisher's PLSD. All analyses were replicated on three independent semen samples except benzoquinone data, which were replicated on nine individual samples.