Abstract
Previous studies have shown that Adjudin (formerly AF2364) is a potential male contraceptive which exerts its effect by detaching spermatids from the seminiferous epithelium in rat testis (Cheng CY et al, 2001, Biol Reprod, 65:449-461). However, the effects of Adjudin on sperm fertilizing ability have not been evaluated. The aims of the present study were to assess the effects of Adjudin on sperm fertilizing ability at nontoxic dose level. Semen samples were obtained from 15 fertile men that were examined following WHO (1999) standard andrology criteria and washed in human fallopian tube fluid (HTF) medium through a discontinuous Percoll gradient (45%-90%). Washed sperm were pre-incubated in HTF medium with or without various concentrations of Adjudin at 10 nM-10 μM. Sperm capacitation and the acrosome reaction (AR) were assessed by using chlortetracyclin staining, and sperm fertilizing ability was evaluated by a sperm penetration of zona-free hamster egg assay. The result showed that sperm capacitation was promoted in the HTF for incubation up to 5 h which was increased in the percentage of ¡°B¡± pattern. However, sperm capacitation was significantly inhibited by AF2364 dose-dependently at concentrations of 10 nM-10μM. In selected experiments, sperm were incubated in HTF with 10 μM Adjudin for 5h, and then washed twice to remove Adjudin and incubated thereafter in Adjudin-free HTF for 1-3h. The percentage of ¡°B¡± pattern was comparable to that of control, indicating that the Adjudin-mediated inhibition of capacitation is reversible. Likewise, Adjudin significantly and dose-dependently inhibited the AR induced by progesterone (15 μM) or rhuZP3 (20 μg/ml). The ARs were inhibited completely by 10 μM Adjudin. However, Adjudin did not affect sperm motility at a concentration of 10 μM. Furthermore, the penetration of spermatozoa into zona-free hamster eggs in the presence of 1 μM Adjudin was significantly decreased, but it did not affect sperm motility as compared to the control. Summary: these results indicate that Adjudin is a potential male contraceptive by preventing sperm capacitation, AR and sperm-egg fusion, but not sperm motility (Supported by grants from the National 973 project 2006CB504002, and NIH NICHD U54 HD029990 Project 5).
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