Voltage-dependent inactivation of inward-rectifying single-channel currents in the guinea-pig heart cell membrane.

Inward currents through single K+ channels in isolated ventricular heart cells of the guinea-pig were recorded using the patch-clamp technique (Hamill, Marty, Neher, Sakmann & Sigworth, 1981). The voltage-dependent gating properties of the channels were examined in the potential range between 0 and -120 mV with 145 mM-KCl on the extracellular side of the membrane patch, i.e. with approximately symmetrical transmembrane K+ concentrations. When voltage pulses from 0 mV to negative test potentials were applied to patches containing several channels, more channels were open at the beginning of the pulses than in the steady state. Averages of many current responses showed inactivation of the mean current in response to the hyperpolarizing voltage pulses. The inactivation was stronger and faster at larger hyperpolarization. The lifetimes of the open and closed states of the channel and the probability of the open state p were estimated from records of the elementary currents at various constant potentials. As indicated by the inactivation of the averaged currents, the value of p was smaller at more negative potentials, approximately 0.15 at -50 mV and 0.02 at -110 mV. This caused a negative slope in the current-voltage relation of the time-averaged current at potentials more negative than -50 mV. The channel openings were grouped in complex bursts. At least three exponentials were needed to fit the frequency histogram of the lifetimes of all closed states (time constants at -50 mV: 1.1 ms, 16 ms and 3.2 s). The lifetimes of the individual openings were exponentially distributed (time constant: 70 ms). The kinetics of the channel were interpreted by two different models involving three states of a channel (closed-closed-open or closed-open-closed). The rate constants and their voltage dependence were estimated for both models. Both models describe the data equally well; the reason for this ambiguity is discussed. The channels are blocked by Cs+ or Ba2+. Cs+ (0.1 mM) caused frequent and short interruptions of the individual channel openings. Ba2+ (0.5 mM) also shortened the openings and in addition decreased the number of openings per burst. The results suggest that the inward-rectifying current IK1 in heart ventricular cells is partially inactivated by hyperpolarization. The inactivation could account for part of the time-dependent decrease in the whole-cell current previously ascribed to depletion of K+.