Partial characterization of human spermatozoal phosphodiesterase and adenylate cyclase and the effect of steroids on their activities.
In sonicated human spermatozoa, phosphodiesterase hydrolyzed adenosine 3':5'-monophosphate (cAMP) at 20.80 +/- 3.23 nmoles/10(8)sperm/20 min at 37 degrees C (50 microM cAMP, initially). In human semen, about 60% of the phosphodiesterase activity was in the spermatozoa. Both polyacrylamide gel electrophoresis and DEAE-cellulose column chromatography indicated that there are at least 5 isozymes of phosphodiesterase. Various steroids were tested at a concentration of 2 micrograms/ml for their effects on phosphodiesterase activity in semen. None was found to have any significant effect. In sonicated human spermatozoa, adenylate cyclase synthesized cAMP at 0.02-2.11 nmoles/10(8)sperm/20 min at 37 degrees C (1 mM ATP, initially) depending on the availability of Mn2+ and caffeine in the assay mixture. Mn2+ was demonstrated to be a potent adenylate cyclase activator in human spermatozoa and its effect on human sperm adenylate cyclase was found to be dose-dependent. Cholera toxin, at a concentration of 20 micrograms/ml, had no effect on human sperm adenylate cyclase activity after it had been incubated with the intact spermatozoa for between 5 min and 5 h at 37 degrees C before the sperm were homogenized and the rate of cAMP formation assayed. In addition, human sperm adenylate cyclase decayed rapidly at 37 degrees C. Of various steroids tested at a concentration of 2 micrograms/ml for their effects on human sperm adenylate cyclase activity, only oestradiol-17 beta showed a significant effect, elevating the rate of cAMP formation by about 30%.
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000378:metabolism*; D000262:Adenylyl Cyclases / Q000378:metabolism*; D002110:Caffeine / Q000494:pharmacology; D002772:Cholera Toxin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004958:Estradiol / Q000494:pharmacology; D006801:Humans; D008297:Male; D008345:Manganese / Q000494:pharmacology; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology*; D013256:Steroids / Q000494:pharmacology*
D013256:Steroids; D002110:Caffeine; D008345:Manganese; D004958:Estradiol; D002772:Cholera Toxin; D000242:Cyclic AMP; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D000262:Adenylyl Cyclases