Abstract
Progestin stimulation of sperm hypermotility remains poorly understood despite having been described in numerous vertebrate species. We show here that progestin stimulation of sperm hypermotility in a teleost, the Atlantic croaker (Micropogonias undulatus) is associated with activation of an olfactory G protein (Golf). Furthermore, we provide evidence that this progestin action is mediated by membrane progestin receptor-alpha (mPRalpha). Golf was identified in croaker sperm membranes and was specifically activated after treatment with the progestin 17,20beta,21-trihydroxy-4-pregnen-3-one (20beta-S). Treatment of sperm membranes with 20beta-S caused an increase in cAMP production, which was blocked by pretreatment with cholera toxin and two membrane adenylyl cyclase inhibitors: 2',5'-dideoxyadenosine and SQ22536. Moreover, preincubation of croaker sperm with 2',5'-dideoxyadenosine and SQ22536 resulted in a significant inhibition of 20beta-S-stimulated hypermotility. Binding of [3H]20beta-S to sperm membranes was decreased after pretreatment with GTPgammaS but not pertussis toxin, suggesting the receptor is coupled to a pertussis toxin-insensitive G protein. Golf and mPRalpha were coexpressed on the sperm midpiece and flagella and were coimmunoprecipitated from sperm membranes. Finally, expression of mPRalpha protein on sperm increased after in vivo treatment with LHRH and was associated with increased induction of sperm motility by 20beta-S. These results suggest that 20beta-S activates mPRalpha in croaker sperm, which in turn activates Golf and membrane adenylyl cyclase to stimulate sperm hypermotility. Taken together these findings provide a plausible mechanism by which progestins stimulate sperm hypermotility in croaker and provide the first evidence of hormonal activation of Golf in any species.