EFFECTS OF 8-BROMOGUANOSINE 3'/5'-CYCLIC MONOPHOSPHATE ON PHENYLEPHRINE-INDUCED PHOSPHATIDYLINOSITOL HYDROLYSIS AND CONTRACTION IN RAT CAUDAL ARTERY

1 The effects of 8-bromoguanosine 3':5'-cyclic monophosphate (8-bromo-cyclic GMP) on phenylephrine-induced contractions and phosphatidylinositol (PI) hydrolysis were investigated in rat isolated caudal artery. The effects of the nucleotide were compared to those of felodipine, a dihydropyridine Ca2+ channel antagonist and ryanodine, a putative depletor of intracellular Ca2+ stores. The purpose of this investigation was to examine the regulatory effects of cyclic GMP on receptor-mediated signal transduction in vascular smooth muscle. 2 Phenylephrine induced a concentration-dependent increase in PI hydrolysis that reached a maximum at 10 mu M phenylephrine. Pre-incubation with felodipine (10 nM) significantly reduced PI turnover, but did not affect basal hydrolysis. Similarly, removal of extracellular Ca2+ (2 mM ethylene glycol-bis(beta-amino-ethyl ether) N, N, N', N'-tetraacetic acid (EGTA)) blocked phenylephrine-induced PI hydrolysis, but did not affect basal turnover. In contrast, 8-bromo-cyclic GMP (10 mu M) did not affect phenylephrine induced PI hydrolysis, nor did it affect basal turnover. 3 Phenylephrine induced concentration-dependent contractions that were inhibited by each of 8-bromo-cyclic GMP (10 mu M), felodipine (1 nM and 10 nM) and ryanodine (3 mu M and 10 mu M). In addition, removal of Ca2+ from the physiological salt solution (2 mM EGTA) completely abolished contractions elicited by phenylephrine. 4 Phenylephrine-induced contractions were not further affected by felodipine and 8-bromo-cyclic GMP applied concomitantly than by equivalent concentrations of felodipine alone. However, ryanodine and 8-bromo-cyclic GMP applied together significantly inhibited: phenylephrine-induced contractions in comparison to ryanodine alone. 5 These results suggest that phosphoIipase C-activated PI hydrolysis in the rat caudal artery is dependent on extracellular Ca2+, mediated, in part, through dihldropyridine-sensitive Ca2+ channels. Inhibition of contraction by felodipine may be brought about through indirect inhibition of IP3 production and subsequent attenuation of intracellular Ca2+ release. 8-Bromo-cyclic GMP does not inhibit PI hydrolysis; it may regulate vascular smooth muscle contraction by inhibition of Ca2+ release from IP3-mediated intracellular stores, but it is unlikely that 8-bromo-cyclic GMP affects ryanodine-sensitive stores.