Possible existence of transient TTX-sensitive chloride current in the membrane of isolated cardiomyocytes

Cardiomyocytes enzymatically isolated from rat and guinea pig ventricular tissue were investigated under conditions of intracellular perfusion and voltage clamp at 18-20 degrees C. Perfusion with 135 mmol/l Tris(HF), pH 7.2 was used to eliminate outward potassium currents. The dependence of inward current (elicited by depolarizing pulses from a holding potential level of--120 mV) on low external TTX concentrations (from 10(-13) to 10(-10) mol/l) was studied. Similar TTX concentrations increased the amplitude of the inward current and changed its kinetics in a large number of cells tested. The effect was fully reversible. The effect could be evaluated in a net form by digital subtraction of the current obtained after the application of a low external TTX concentration from the initial current in a TTX-free solution. The TTX concentration dependence of the difference current could be fitted by one-to-one binding curve with Kd = (1.0 +/= 0.4) x 10(-12) mol/l. TTX-induced current changes were absent in low sodium or chloride-free external solutions. The outward current (a block of which by TTX produced the inward current changes observed) showed a reversal potential consistent with the chloride nature of such a current. The existence of a transient TTX-sensitive Na-dependent potential gated chloride current in the membrane of isolated cardiomyocytes is postulated.