Problem: Nitric oxide (NO) has been known to have multifunctional roles both in the male and female reproductive systems. We investigated the effects of sodium nitroprusside (SNP)-dependent NO release on sperm cell function and embryo development to elucidate the mechanisms of action of NO.
Method of study: Semen samples from 20 healthy men were processed by the swim-up method. Sperm motility, hyperactivation, and acrosome reaction were examined following incubation with various concentrations of SNP. The concentration of 10 nM to 1 mM was used for sperm motility and hyperactivation measurement and 1 microM to 1 mM for examining the effect on acrosome reaction. Embryo development to blastocyst stage was assessed using 100 nM to 1 mM of SNP added before transferring the mouse embryos into the culture medium. Finally, to understand the mechanism of action of NO, changes in embryo development were examined after zygotes were treated with various concentrations ranging up to 1 mM of 8-bromo-cGMP, an analog of cGMP.
Results: Both sperm motility and hyperactivation were significantly reduced at 100 microM and 1 mM concentrations of SNP after 6 hr of incubation. After 24 hr of incubation, they were greatly decreased with all, except the 10 nM concentration of SNP. The percentage of acrosomal-reacted spermatozoa was increased with the increasing concentration of SNP following incubation with 10 microM and 1 mM of SNP. Embryo development was arrested since the two-cell embryonic stage with all except the 100 nM concentration of SNP, and inhibited by 200 microM of SNP regardless of SNP treatment stage. However, embryo development was not influenced by 8-bromo-cGMP.
Conclusions: We concluded that SNP-inhibited sperm cell function and embryo development in a dose- and time-dependent manner, and the inhibitory effect on embryo development, may not be a stage-specific treatment mediated via a cGMP-independent pathway. This result suggests that NO may be enough to affect the fecundity potential in vivo.