Vaginal fluid adenosine 3',5'-cyclic monophosphate (cAMP) in the rat: interaction with sperm cAMP-dependent protein...

Cyclic AMP (adenosine 3',5'-cyclic monophosphate) concentrations were determined in rat vaginal fluids throughout the estrous cycle. Radioimmunoassay results demonstrated that estrus and early metestrus vaginal fluids had significantly (p less than 0.01) elevated cAMP concentrations compared to proestrus, late metestrus, and early and late diestrus. Ovariectomy reduced RIA-detectable cAMP in vaginal fluid. When cauda sperm were preincubated for 5 min with vaginal fluids from each stage of the estrous cycle, results demonstrated that only estrus- and early metestrus-stage vaginal fluids caused a decrease in [32P]-8N3cAMP (8-azido photoaffinity analogue of cAMP) photolabeling of sperm cAMP-dependent protein kinase regulatory subunits RI and RII. To examine if this reduction in [32P]-8N3cAMP photoincorporation by sperm RI and RII could be due to endogenous cAMP, vaginal fluids were boiled, trypsinized, and/or incubated with EGTA or phosphodiesterase. Only phosphodiesterase-treated vaginal fluids restored sperm regulatory subunit photoincorporation of [32P]-8N3cAMP. It is suggested that cAMP is present in rat vaginal fluids during the estrous cycle in a concentration sufficient to bind the regulatory subunits of rat sperm cAMP-dependent protein kinase.