Evidence for modulation of progesterone secretion by calcium and protein kinase C activators in ovine chorionic cells.
The hypothesis that calcium-dependent mechanisms may be involved in regulating ovine placental steroidogenesis was investigated using chorionic cells isolated by enzymatic digestion. Treatment of the cells with the calmodulin antagonist trifluoperazine (TFP) or pimozide caused a dose-related inhibition of progesterone (P4) production by 80 percent (P less than 0.001) at 40 microM TFP and 56 per cent (P less than 0.001) at 10 microM pimozide. Moreover, the conversion of 25 hydroxycholesterol (25 OH Chol.) to P4 was impaired in the presence of these compounds. These experiments suggest the involvement of a calcium-calmodulin system in the regulation of ovine placental P4 synthesis. Interestingly, calcium ionophore A23187 caused a gradual decline in P4 secretion and completely blocked it at 1 microM (P less than 0.001) and remains absent even in the presence of 25 OH Chol. In contrast, EGTA increased P4 secretion (P less than 0.01). Further, in the presence of 3 mM EGTA the inhibitory effect of 1 microM A23187 was fully reversed. Taken together these results suggest that extracellular calcium could play a role of negative modulation of P4 secretion in these cells. The possible involvement of protein kinase C (PKC) was tested using tumor-promoting phorbol ester (PMA) or permeant diacylglycerols (OAG or DOG). These compounds were unable to modify basal P4 secretion but reduced 25 OH Chol stimulated secretion to basal level. The phorbol ester that was unable to activate PKC had no effect on the metabolism of 25 OH chol. Thus, PMA and diacylglycerol effects are probably mediated by PKC. These data support the hypothesis that PKC activation plays a role in the modulation of cholesterol side-chain cleavage activity in ovine chorionic cells. These results show that calcium-dependent processes are involved in both positive and negative control of P4 secretion by ovine placenta. Our results also suggest a role for calmodulin and PKC pathways in modulating this secretion.
de la Llosa-Hermier
M P
MP
C.N.R.S., Laboratoire de Biochimie des Hormones, Gifsur-Yvette, France.
Martal
J
J
Ricour
A
A
Hermier
C
C
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002823:Chorion / Q000187:drug effects / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D006888:Hydroxycholesterols / Q000494:pharmacology; D066298:In Vitro Techniques; D011247:Pregnancy; D011374:Progesterone / Q000096:biosynthesis / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D012756:Sheep; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D006888:Hydroxycholesterols; D000001:Calcimycin; D011374:Progesterone; C007997:25-hydroxycholesterol; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium