Tumor promoters as probes of protein kinase C in dog thyroid cell: inhibition of the primary effects of carbamylocholine and reproduction of some distal effects.
The acute effects of phorbol esters, used as probes of protein kinase C activation, were studied on dog thyroid slices incubated in vitro. The derivatives used were: tetradecanoylphorbol acetate (TPA), phorbol-12,13, didecanoate (PDD), phorbol-12,13-diacetate (PDA), and phorbol dibutyrate (PDBu) and as inactive controls, phorbol itself, phorbol-12, myristate and phorbol-13, acetate, in concentrations ranging from 5.10(-8) to 5.10(-6) mol/L. The active phorbol esters had no effect on basal cyclic AMP concentrations; they inhibited cyclic AMP accumulation induced by prostaglandin E1 but not that induced by thyrotropin (TSH) 1 mU/mL and forskolin 10 mumol/L. Phorbol esters like carbamylcholine acutely stimulated iodide organification and inhibited the stimulation of hormone secretion resulting from TSH, Cholera Toxin, forskolin, and Bu2-cyclic AMP action. These metabolic effects did not require the presence of extracellular Ca++, and could not be antagonized by Ca++ depletion or manganese addition. The active phorbol esters abolished the cyclic AMP independent increased PI turnover induced by TSH 10 mU/mL or carbamylcholine (Cchol) 10(-6) mol/L but did not affect the basal incorporation of 32P into phosphatidylinositol. They reduced the 45Ca efflux from preloaded slices below basal levels and blocked the increased 45Ca release induced by TSH and Cchol. They also inhibited the increase in cyclic GMP concentrations resulting from Cchol action but not the effect of the ionophore A23187 (10(-5) mol/L) nor the basal levels of cyclic GMP.(ABSTRACT TRUNCATED AT 250 WORDS)
D000527:Alprostadil / Q000494:pharmacology; D000818:Animals; D002217:Carbachol / Q000037:antagonists & inhibitors*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004075:Diglycerides / Q000494:pharmacology; D004285:Dogs; D010703:Phorbol Esters / Q000494:pharmacology*; D010716:Phosphatidylinositols / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013961:Thyroid Gland / Q000187:drug effects / Q000201:enzymology*; D013972:Thyrotropin / Q000494:pharmacology
D004075:Diglycerides; D010703:Phorbol Esters; D010716:Phosphatidylinositols; D005576:Colforsin; D002217:Carbachol; D013972:Thyrotropin; D000242:Cyclic AMP; D011493:Protein Kinase C; D000527:Alprostadil; D013755:Tetradecanoylphorbol Acetate