Regulation of the corpus luteum by protein kinase C. I. Phosphorylation activity and steroidogenic action in large and small ovine luteal cells.
The activity and steroidogenic action of protein kinase C were evaluated in small and large steroidogenic ovine luteal cells. Protein kinase C activity (per mg protein) was threefold greater in large than in small luteal cells, whereas protein kinase A activity was similar in the two cell types. Phorbol 12-myristate 13-acetate (PMA) activated protein kinase C in luteal cells as demonstrated by membrane association of 91% of available protein kinase C within 15 min of PMA treatment. Longer treatments with PMA produced cells with low protein kinase C activity (protein kinase C-deficient cells) but did not affect cellular viability or protein kinase A activity. Activation of protein kinase C caused an acute, dose-dependent inhibition of progesterone production in unstimulated large and luteinizing hormone (LH)-stimulated small luteal cells. This inhibition by PMA appeared to be specific for protein kinase C since it was greatly attenuated in protein kinase C-deficient cells and since an inactive phorbol ester, 4 alpha-phorbol, had no effect on luteal progesterone production. The inhibitory locus of protein kinase C action in small luteal cells appeared to be distal to the adenylate cyclase enzyme because progesterone production was inhibited similarly in cells stimulated with LH, forskolin, or dibutyryl cyclic adenosine 3',5'-monophosphate. Cholesterol side-chain cleavage activity, as measured by metabolism of 25-hydroxycholesterol, was inhibited by PMA in large, but not in small, luteal cells. These data indicate that activation of protein kinase C specifically inhibits progesterone production in both large and small ovine luteal cells, although the intracellular mechanisms invoked appear to differ in the two cell types.
Department of Physiology, Colorado State University, Fort Collins 80523.
M C
MC
Wiltbank
J J
JJ
Knickerbocker
G D
GD
Niswender
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003338:Corpus Luteum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D066298:In Vitro Techniques; D010743:Phospholipids / Q000494:pharmacology; D010766:Phosphorylation; D011374:Progesterone / Q000096:biosynthesis*; D011493:Protein Kinase C / Q000502:physiology*; D011863:Radioimmunoassay; D012756:Sheep; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D010743:Phospholipids; D011374:Progesterone; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium