Figure 1.

Atorvastatin effects on membrane cholesterol distribution, P-Tyr and acrosome reaction in spermatozoa. Membrane cholesterol distribution, P-Tyr and acrosome integrity were analysed in spermatozoa obtained from 17 normocholesterolaemic men before, during and 3 months after the end of treatment as described in Methods. P-Tyr patterns were assessed before (t = 0 h) and after (t = 3 h) incubating spermatozoa in capacitating conditionsd and visualized by western blotting using a monoclonal anti-phosphotyrosine antibody. To calibrate the signal for the amount of sperm protein, the same membranes were reprobed using a monoclonal anti-alpha-tubulin antibody. Acrosome integrity was assessed before (5 min) and after 3 h of incubation in capacitating conditions (3 h), by epifluorescence microscopy (x400) using PSA-FITC as a probe. A represents mean ± SEM of the percentage of sperm cells showing a redistribution of cholesterol with filipin labelling absent in the sperm head. Cholesterol distribution was estimated by epifluorescence microscopy (x400) using filipin as a probe. B represents typical patterns of protein tyrosine phosphorylation (P-Tyr) and α-tubulin in human spermatozoa. C represents P-Tyr signal normalized to the tubulin signal and the ratios were related to the basal signal obtained before incubation. Data are represented as mean ± SEM in arbitrary units (a.u.). D represents a fluorescence micrograph showing a sperm cell with an intact acrosome membrane (A pattern: marked fluorescence in the acrosome region) and a sperm cell without an acrosome membrane (AR pattern: no fluorescence or marked fluorescence along the equatorial segment). E represents mean ± SEM of the percentage of AR spermatozoa. *indicates values significantly different from those measured before atorvastatin intake with p < 0.05.