Systolic [Ca(2+)](i) regulates diastolic levels in rat ventricular myocytes

KEY POINTS: For the heart to function as a pump, intracellular calcium concentration ([Ca(2+)](i)) must increase during systole to activate contraction and then fall, during diastole, to allow the myofilaments to relax and the heart to refill with blood. The present study investigates the control of diastolic [Ca(2+)](i) in rat ventricular myocytes. We show that diastolic [Ca(2+)](i) is increased by manoeuvres that decrease sarcoplasmic reticulum function. This is accompanied by a decrease of systolic [Ca(2+)](i) such that the time‐averaged [Ca(2+)](i) remains constant. We report that diastolic [Ca(2+)](i) is controlled by the balance between Ca(2+) entry and Ca(2+) efflux during systole. The results of the present study identify a novel mechanism by which changes of the amplitude of the systolic Ca transient control diastolic [Ca(2+)](i). ABSTRACT: The intracellular Ca concentration ([Ca(2+)](i)) must be sufficently low in diastole so that the ventricle is relaxed and can refill with blood. Interference with this will impair relaxation. The factors responsible for regulation of diastolic [Ca(2+)](i), in particular the relative roles of the sarcoplasmic reticulum (SR) and surface membrane, are unclear. We investigated the effects on diastolic [Ca(2+)](i) that result from the changes of Ca cycling known to occur in heart failure. Experiments were performed using Fluo‐3 in voltage clamped rat ventricular myocytes. Increasing stimulation frequency increased diastolic [Ca(2+)](i). This increase of [Ca(2+)](i) was larger when SR function was impaired either by making the ryanodine receptor leaky (with caffeine or ryanodine) or by decreasing sarco/endoplasmic reticulum Ca‐ATPase activity with thapsigargin. The increase of diastolic [Ca(2+)](i) produced by interfering with the SR was accompanied by a decrease of the amplitude of the systolic Ca transient, such that there was no change of time‐averaged [Ca(2+)](i). Time‐averaged [Ca(2+)](i) was increased by β‐adrenergic stimulation with isoprenaline and increased in a saturating manner with increased stimulation frequency; average [Ca(2+)](i) was a linear function of Ca entry per unit time. Diastolic and time‐averaged [Ca(2+)](i) were decreased by decreasing the L‐type Ca current (with 50 μm cadmium chloride). We conclude that diastolic [Ca(2+)](i) is controlled by the balance between Ca entry and efflux during systole. Furthermore, manoeuvres that decrease the amplitude of the Ca transient (without decreasing Ca influx) will therefore increase diastolic [Ca(2+)](i). This identifies a novel mechanism by which changes of the amplitude of the systolic Ca transient control diastolic [Ca(2+)](i).