High carbachol increases the electrically induced [Ca2+]i transient in the single isolated ventricular myocyte of...

In order to investigate the mechanisms responsible for the inotropic effects of muscarinic acetylcholine receptor stimulation by high concentrations of muscarinic receptor agonists, we studied the effects of carbachol at 30-300 microM on the electrically induced [Ca2+]i transient of rat isolated ventricular myocytes. Carbachol at this dose range increased the amplitude and duration of the electrically induced [Ca2+]i transient time and dose dependently. It also increased the resting fluorescence ratio and time to 80% decline of amplitude from the peak. At 100-300 microM the increase in [Ca2+]i transient was followed by a cluster of Ca2+ oscillations in 50-83% of the cells studied. The effects were blocked by atropine, but not pertussis toxin. Depletion of Ca2+ from sarcoplasmic reticulum by ryanodine, which itself reduced the amplitude of the [Ca2+]i transient and increase resting fluorescence, abolished the effect of carbachol on the [Ca2+]i transient without affecting its effect on resting fluorescence ratio. The caffeine-induced [Ca2+]i transient was unaffected by prior addition of carbachol in a Ca2+ free and low Na+ solution. Inhibition of Ca2+ by the L-type Ca2+ channel blocker, verapamil, which itself reduced the amplitude of the [Ca2+]i transient without affecting the resting fluorescence ratio, attenuated the augmentation of the amplitude of the [Ca2+]i transient elicited by carbachol. Ni2+, a non-specific Ca2+ channel blocker and an inhibitor of Na(+)-Ca2+ exchange, abolished the effects of carbachol on both [Ca2+]i transient and resting fluorescence ratio. Low external Na+, which increased the resting fluorescence ratio due to its inhibitory effect on Na(+)-Ca2+ exchange, also abolished the effects of carbachol. The results indicate that the inotropic effect of muscarinic acetylcholine receptor stimulation by high concentrations of a muscarinic receptor agonist may be due to an increase in the electrically induced [Ca2+]i transient in ventricular myocytes via a process which is not pertussis toxin sensitive. The increase in the electrically induced [Ca2+]i transient may result from increases in Na2(+)-Ca2+ exchange and influx of Ca2+ via voltage-gated Ca2+ channels, and mobilization of Ca2+ from the intracellular store. The mobilization of Ca2+ from the intracellular store is a secondary event. The study has provided for the first time that muscarinic acetylcholine receptor stimulation by high concentrations of carbachol increases Ca2+ influx via the Ca2+ channel and mobilization of Ca2+ from its intracellular store. The study has also demonstrated for the first time the occurrence of Ca2+ oscillations induced by high concentrations of carbachol.