The effects of membrane potential, extracellular potassium, and tetrodotoxin on the intracellular sodium ion activity of sheep cardiac muscle.

The intracellular sodium ion activity was measured using liquid ion-exchange microelectrodes with rapid response times in sheep Purkinje fibers and ventricular muscle under voltage control. The mean sodium ion activity in quiescent Purkinje fibers was 8.5 mM at a holding potential of -80 mV. With maintained hyperpolarizing (-110 mV) or depolarizing (-40 and 0 mV) voltage steps, sodium ion activity increased or decreased, respectively. At 0 mV, the mean steady state value for the sodium ion activity was 3.8 mM. Following a voltage step to 0 mV, or back to -80 mV, the time course of the sodium ion activity change could be fitted by single exponentials, with similar half-times. Increasing the extracellular potassium ion concentration from 5.4 to 15 mM did not alter the steady state value of the sodium ion activity at clamped voltages of -80 or 0 mV, which suggests that the external potassium ion activating site of the Na-K pump was saturated. With the extracellular potassium concentration 0 mM (holding potential -80 mV), the sodium ion activity increased. When maintained depolarizing steps to 0 mV were applied, the sodium ion activity decreased by up to 20 mM. This large fall in sodium ion activity is assumed to represent partial reactivation of the Na-K pump due to potassium ion accumulation in clefts. We also studied the stimulation-dependent change in sodium ion activity. Trains of action potentials or short duration depolarizing voltage clamp steps caused a frequency dependent rise in sodium ion activity. The magnitude of the rise of sodium ion activity was not altered by lengthening the duration of each voltage clamp step, but was inhibited by tetrodotoxin or by holding the membrane potential at -50 mV between depolarizing steps. These results show that sodium ion activity is a complex function of membrane voltage, depolarization frequency, and time. The rise in sodium ion activity with stimulation appears to depend on sodium ion entry regulated by the sodium channel, and may be important in the modulation of intracellular calcium and tension through the Na+-Ca++ exchange mechanism.