Dependence of cyclic-AMP induced relaxation on Ca2+ and calmodulin in skinned smooth muscle of guinea pig Taenia coli.

Abstract

cAMP (10(-6) - 10(-4) M) produced a dose-dependent relaxation of Ca2+-induced contraction in the guinea-pig taenia coli skinned with 1% Triton X-100. At 0.53 microM Ca2+ and 0.05 microM calmodulin (CaM), cAMP (10(-4) M) produced a maximal relaxation of 75% (pH 6.7; 25 degrees C). Increasing Ca2+ (0.8 microM) or CaM (0.37 microM) reduced cAMP-induced relaxation to 25 and 5% respectively. At high CaM (5 microM), cAMP-induced relaxation could be completely inhibited by as low as 0.25 microM Ca2+. Furthermore, small increases in Ca2+ or CaM could effectively reverse the cAMP-induced relaxation in the continuous presence of cAMP. These results demonstrate that small modulations in the Ca2+-calmodulin activity have a strong effect on the ability of cAMP to produce a direct relaxing effect on the contractile proteins in skinned fiber. It is suggested that the effects of cAMP on the cellular mechanisms that lower cytoplasmic free Ca2+ concentration may act as the important determinants of the extent of the direct inhibitory effect of cAMP on the contractile elements. These two mechanisms may act in concert in this fashion to effect cAMP-induced relaxation in smooth muscle during beta-adrenergic stimulation.