Modulation of excitation–contraction coupling by isoproterenol in cardiomyocytes with controlled SR Ca2+ load and Ca2+ current trigger

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Abstract
Cardiac Ca2+ transients are enhanced by cAMP‐dependent protein kinase (PKA). However, PKA‐dependent modulation of ryanodine receptor (RyR) function in intact cells is difficult to measure, because PKA simultaneously increases Ca2+ current (ICa), SR Ca2+ uptake and SR Ca2+ loading (which independently increase SR Ca2+ release). We measured ICa and SR Ca2+ release ± 1 μm isoproterenol (ISO; isoprenaline) in voltage‐clamped ventricular myocytes of rabbits and transgenic mice (expressing only non‐phosphorylatable phospholamban). This mouse model helps control for any effect of ISO‐enhanced SR uptake on observed release, but the two species produced essentially identical results. SR Ca2+ load and ICa were adjusted by conditioning. We thus evaluated PKA effects on SR Ca2+ release at constant SR Ca2+ load and ICa trigger (with constant unitary ICa). The amount of SR Ca2+ release increased as a function of either ICa or SR Ca2+ load, but ISO did not alter the relationships (measured as gain or fractional release). This was true over a wide range of SR Ca2+ load and ICa. However, the maximal rate of SR Ca2+ release was ∼50% faster with ISO (at most loads and ICa levels). We conclude that the isolated effect of PKA on SR Ca2+ release is an increase in maximal rate of release and faster turn‐off of release (such that integrated SR Ca2+ release is unchanged). The increased amount of SR Ca2+ release normally seen with ISO depends primarily on increased ICa trigger and SR Ca2+ load, whereas faster release kinetics may be the main result of RyR phosphorylation.