Cyclic GMP-dependent protein kinase activation mediates inhibition of catecholamines secretion and Ca2+ influx in bovine chromaffin cells.
The effects of the membrane-permeable cGMP analogue, 8-bromoguanosine 3':5'-cyclic monophosphate on acetylcholine-evoked catecholamine secretion and cytosolic calcium increases were studied in chromaffin cells from the bovine adrenal gland. Preincubation with 100 microM 8-bromoguanosine 3':5'-cyclic monophosphate during 10 and 30 min decreased the acetylcholine-evoked catecholamine release by 16 +/- 3% and 27 /+- 5%, respectively. The cytosolic calcium increases triggered by acetylcholine and 30 mM KCl were also inhibited by 30 min of preincubation with this compound by 27 +/- 4 and 34 /+- 12%, respectively. Changes in membrane potential induced by acetylcholine and KCl were not affected by preincubation with 8-bromoguanosine 3':5'-cyclic monophosphate. The cyclic GMP-dependent protein kinase inhibitor N-[2-(methylamino)ethyl]-5-isoquinoline sulfonamide dihydrochloride-at l micron abolished the inhibitory effect of 8-bromoguanosine 3':5'-cyclic monophosphate on acetylcholine-evoked calcium increase. By contrast, a potent and selective inhibitor against cyclic AMP-dependent protein kinase, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide did not block the 8-bromoguanosine 3':5'-cyclic monophosphate effect. Additionally, 8-bromoguanosine 3':5'-cyclic monophosphate stimulated histone F2b phosphorylation by a partial purified cGMP-dependent protein kinase from chromaffin cells. The extent of histone phosphorylation was reduced by N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride and 8-(4-chlorophenylthio)-guanosine 3':5'-cyclic monophosphorothioate, Rp-isomer, a specific inhibitor against cyclic GMP-dependent protein kinase, whereas it was not modified by N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinoline sulfonamide. The results suggest that the inhibitory effects of 8-bromoguanosine 3':5' cyclic monophosphate on chromaffin cells are mediated through the activation of cGMP-dependent protein kinase.(ABSTRACT TRUNCATED AT 250 WORDS)
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002395:Catecholamines / Q000378:metabolism*; D002417:Cattle; D002838:Chromaffin System / Q000187:drug effects / Q000378:metabolism*; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D003864:Depression, Chemical; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects
D002395:Catecholamines; C016276:8-bromocyclic GMP; D003994:Bucladesine; D017869:Cyclic GMP-Dependent Protein Kinases; D006152:Cyclic GMP; D000109:Acetylcholine; D002118:Calcium