Physiologic gating properties of unitary cardiac L-type Ca2+ channels

Abstract

The contraction of adult mammalian ventricular cardiomyocytes is triggered by the influx of Ca2+ ions through sarcolemmal L-type Ca2+ channels (LCCs). However, the gating properties of unitary LCCs under physiologic conditions have remained elusive. Towards this end, we investigated the voltage-dependence of the gating kinetics of unitary LCCs, with a physiologic concentration of Ca2+ ions permeating the channel. Unitary LCC currents were recorded with 2mM external Ca2+ ions (in the absence of LCC agonists), using cell-attached patches on K-depolarized adult rat ventricular myocytes. The voltage-dependence of the peak probability of channel opening (Po vs. Vm) displayed a maximum value of 0.3, a midpoint of −12mV, and a slope factor of 8.5. The maximum value for Po of the unitary LCC was significantly higher than previously assumed, under physiologic conditions. We also found that the mean open dwell time of the unitary LCC increased twofold with depolarization, ranging from 0.53±0.02ms at −30mV to 1.08±0.03ms at 0mV. The increase in mean LCC open time with depolarization counterbalanced the decrease in the single LCC current amplitude; the latter due to the decrease in driving force for Ca2+ ion entry. Thus, the average amount of Ca2+ ions entering through an individual LCC opening (∼300–400 ions) remained relatively constant over this range of potentials. These novel results establish the voltage-dependence of unitary LCC gating kinetics using a physiologic Ca2+ ion concentration. Moreover, they provide insight into local Ca2+-induced Ca2+ release and a more accurate basis for mathematical modeling of excitation–contraction coupling in cardiac myocytes.