Regulation of force and intracellular calcium transients by cyclic AMP generated by forskolin, MDL 17,043 and isoprenaline, and its modulation by muscarinic receptor agents: a novel mechanism for accentuated antagonism.

The relation of changes in intracellular calcium transients and force of isometric contractions in response to an elevation or reduction of cyclic AMP levels was investigated in isolated dog ventricular trabeculae and rabbit papillary muscles, in which multiple superficial cells have been microinjected with the calcium sensitive bioluminescent protein aequorin. Forskolin, MDL 17,043 and isoprenaline elevated the tissue cyclic AMP level, increased consistently the peak aequorin signals and force, and abbreviated the duration of both signals in a concentration-dependent manner. When the effect of isoprenaline was compared with that of alteration of extracellular calcium concentration [( Ca2+]0), the increase in force by isoprenaline was associated with higher peak aequorin signals than that by alteration of [Ca2+]0 for a given increase in force, indicating the decrease in calcium sensitivity of myofibrils by cyclic AMP generated by beta-adrenoceptor stimulation. Carbachol, which did not affect significantly the basal force and cyclic AMP levels, lowered the cyclic AMP levels elevated previously by forskolin, MDL 17,043 or isoprenaline in the isolated dog ventricular trabeculae. It antagonized the increase in peak aequorin signals and force caused by these agents in a concentration-dependent manner. When carbachol had been administered prior to isoprenaline and the concentration-response curve for isoprenaline was determined in the presence of carbachol, the relation of force peak aequorin signals was not modified by carbachol in the rabbit papillary muscle. Carbachol, when administered during induction of the positive inotropic action by forskolin, MDL 17,043 and isoprenaline, decreased the force more than peak aequorin signals in a concentration-dependent manner in the dog ventricular trabeculae. Therefore, the relation of force to peak aequorin signals was shifted downwards during the carbachol-induced inhibition, indicating a further decrease of calcium sensitivity of myofibrils by carbachol. This effect of carbachol appears to be specific to the cyclic AMP-mediated positive inotropic action, since the alpha-adrenoceptor-mediated (cyclic AMP-independent) action was unaffected by carbachol. This mechanism may play an important role for "accentuated antagonism" in the mammalian ventricular myocardium.