Cellular mechanisms mediating the stimulation of ovine endometrial secretion of prostaglandin F2α in response to oxytocin: role of phospholipase C and diacylglycerol

Abstract
The first objective was to describe and evaluate the relationship between the ability of oxytocin to stimulate the activity of phospholipase (PL) C and its ability to stimulate the release of prostaglandin (PG) F in ovine endometrial tissue. Caruncular endometrial tissue was collected from ovariectomized ewes after completion of an 11-day steroid replacement protocol. In experiment 1, explants were incubated either in the presence (10−6 m) or absence of oxytocin for 0, 1, 3, 10, 30 or 100 min to examine the time-course for activation of PLC and release of PGF in response to oxytocin. An increase in the activity of PLC was detected at 3 min while an increase in the release of PGF was not detected until 10 min (P−6 m) to compare their abilities to activate PLC and release PGF. Oxytocin and three receptor angonists stimulated the activity of PLC and the release of PGF (P−9 to 10−6 m to establish dose–response curves for the activation of PLC and release of PGF. For both hormones, significant increases (P2α were observed at 10−8 m while increases in PLC activity were not detected until 10−7 m was used. In experiment 4, explants were pretreated with either U-73122 (an inhibitor of PLC activity) or U-73343 (an inactive analogue of U-73122). Explants were then treated with control medium, oxytocin or AlF4. Both oxytocin and AlF4 stimulated the activity of PLC and the release of PGF (P2α (P0·1). Based on the results from these experiments, the role of PLC in mediating the stimulatory effect of oxytocin on the release of PGF remains unclear. The second objective was to evaluate the role of diacylglycerol (DAG) in mediating the stimulatory effect of oxytocin on endometrial secretion of PGF. In experiment 5, explants were incubated in vitro with varying doses of two DAG analogues. Both analogues stimulated the release of PGF at 10−6 m (P2α. The synthetic DAGs were less effective in stimulating the release of PGF at the doses tested. In experiment 7, explants were preincubated with R59022 or LiCl. R59022 enhanced both the basal and oxytocin-stimulated released of PGF (P=0·07). LiCl promoted an increase in the accumulation of inositol trisphosphate (P0·5). These data indicate that DAG stimulates release of PGF from ovine endometrial tissue and may mediate the stimulatory effect of oxytocin on release of PGF. Journal of Endocrinology (1994) 141, 481–490

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