Interactions of protein kinase C with receptor- and non-receptor-mediated cyclic AMP generation in swine granulosa cells.
Interactions between signal transducing systems may be important in the integrated control of cellular processes in basal and hormonally regulated cells. The swine granulosa cell provides a model to study the interactions between the cAMP and calcium-lipid-dependent signaling pathways. To this end, porcine granulosa cells were incubated in monolayer culture for 1-4 days in the presence of FSH (200 ng/ml), forskolin (85 microM), or cholera toxin (3 micrograms/ml) with or without an activator of protein kinase C, the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) (30 ng/ml). TPA had little effect on basal cAMP generation (1-4 days) or on follicle-stimulating hormone (FSH)-stimulated cAMP formation during the first 24 h. Phorbol ester did inhibit cAMP formation on day 2 (by approximately 25%), on day 3 (by approximately 70%) and on day 4 (by greater than 80%). Forskolin-mediated cAMP generation was inhibited (33-56%) on days 1-4, respectively. TPA suppressed dose-dependent FSH (3-300 ng/ml)-stimulated cAMP production on day 2, virtually abolished FSH-provoked cAMP formation on day 4 and inhibited dose-dependent forskolin-stimulated cAMP production on both days. TPA had no effect on the half-maximally effective dose, ED50, of FSH-stimulated cAMP production but did decrease the ED50 of forskolin and the maximal stimulatory effect of FSH and forskolin on days 2 and 4. Similar effects were observed with the synthetic diacylglycerols DOG (1,2-dioctanoylglycerol) and OAG (1-oleoyl-2-acetylglycerol). The TPA effect was limited to the mammalian adenylate cyclase as it had no effect on bacterially derived adenylate cyclase from Bordetella pertussis.(ABSTRACT TRUNCATED AT 250 WORDS)
Wheeler
M B
MB
Department of Internal Medicine, University of Virginia School of Medicine, Charlottesville 22908.
Veldhuis
J D
JD
D000818:Animals; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis*; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000378:metabolism; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000502:physiology*; D015398:Signal Transduction; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004075:Diglycerides; D011956:Receptors, Cell Surface; D005576:Colforsin; D005640:Follicle Stimulating Hormone; D002772:Cholera Toxin; D000242:Cyclic AMP; D011493:Protein Kinase C; D019204:GTP-Binding Proteins; D013755:Tetradecanoylphorbol Acetate