Ca2+ flux and beating in leaky heart cells.

Previous work has shown that beating heart muscle cells with leaky sarcolemmae take up Ca2+ from the medium at a rate of 5.4 nmol/min/mg of protein while beating at a rate of 44 b.p.m. In the present work, we have used fragments of myocardium (MF), composed of such cells, to measure Ca2+ effux velocity and to compare influx and efflux rates to contraction frequency. The MF were estimated to be three cells thick, five cells wide, and three cells long, on the average. With MF suspended in fresh Pi-buffered medium containing 8.7 mumol/liter total Ca2+, the initial velocity of Ca2+ uptake (Vi) was much greater than the initial velocity of efflux (Vo). Vi, but not Vo, covaried with beating as a function of temperature and also showed ATP dependence. Thus, uptake, but not efflux, is a controlled process coupled to beating under these conditions. When cells were preloaded with Ca2+ and resuspended in Ca2+-depleted medium (total Ca2+ about 1 mumol/liter), approximating the steady state condition, Vi was reduced while Vo increased proportionally. These data suggest that contraction-activating Ca2+ is derived from extracellular sources during the pre-steady state conditions used here. Derivation from intracellular sites could occur in the steady state. The pre-steady state results conflict with mechanical behavior studies by us and others and, with Ca2+ flux in isolated sarcoplasmic reticulum (SR). The steady state results suggest that this conflict may be due to differences in Ca2+ loading and [Ca2+]i/[Ca2+]o.