Role of sodium-calcium exchange in activation of contraction in rat ventricle

1. The functional role of reverse Na(+)-Ca2+ exchange in the activation of contraction of rat ventricular myocytes has been studied. Mechanical activity of single cells, measured as unloaded cell shortening, was recorded simultaneously with membrane current and voltage using a single microelectrode voltage clamp and a video edge detection device. 2. The voltage dependence of contraction was studied by applying trains of depolarizations. At test potentials between +20 and +80 mV (under conditions where large outward currents were activated) a plateau on the shortening vs. voltage (S-V) relationship was observed. Significant cell shortening also occurred at test potentials between -70 and -40 mV; and these contractions were accompanied by large inward Na+ currents. We have investigated the ionic mechanisms for three components of the S-V relation in rat ventricle: (i) shortening which occurs between -70 and -40 mV and is thought to be dependent on the sodium current; (ii) phasic contractions in the voltage range -40 to +40 mV where the L-type Ca2+ current is present; (iii) the plateau of the S-V relation at strongly depolarized voltages where reverse Na(+)-Ca2+ exchange may occur. 3. Experiments in which two independent microelectrode impalements were made in a single myocyte showed that during activation of contraction at test potentials between -70 and -40 mV, and during very large depolarizations (+20 to +80 mV), there were significant deviations of the measured membrane potential from the applied voltages. Activation of cell shortening in these voltage ranges could be eliminated by electronic series resistance compensation, which significantly reduced these voltage errors. Consistent with these findings, when tetrodotoxin (TTX) and 4-aminopyridine (4-AP) were used to block inward Na+ and transient outward K+ currents, respectively, no significant voltage errors were present and a bell-shaped shortening-voltage (S-V) relationship was obtained. 4. When Na+ and K+ currents were blocked, depolarizations from holding potentials of either -80 or -50 mV demonstrated that the threshold for activation of contraction was about -30 mV, and that the voltage dependence of peak shortening was very similar to that of the L-type Ca2+ current (ICa,L). These contractions were suppressed completely by either Cd2+ or ryanodine, showing that activation of cell shortening was due to Ca2+ influx through L-type channels which induced release of Ca2+ from the sarcoplasmic reticulum (SR). No T-type calcium currents were observed.(ABSTRACT TRUNCATED AT 400 WORDS)