1,25(OH)2D3 effects on motility and acrosin activity are VDR-mediated. Percoll-purified sperm washed spermatozoa were incubated in the unsupplemented Earle's medium for 30 min at 37°C and 5% CO2, in the absence (NC) or in the presence of increasing 1,25(OH)2D3 concentrations (0.01 nM, 0.1 nM, 1 nM and 10 nM) or with 0.1 nM 1,25(OH)2D3 combined with anti-VDR Ab (AbVDR). Other samples were incubated in capacitating medium (Cap). Sperm motility and acrosin activity were assessed as reported in Materials and methods. The sperm motility presented are representative examples of experiments that were performed at least five times with repetitive results while acrosin activity were performed at least seven times with repetitive results. Columns are mean ± S.D. Data are expressed as % for motility and as μIU acrosin/106 sperms for acrosin activity. *P < 0.05 versus control; ** P < 0.02, ***P < 0.01 versus control.