Abstract
The effect of tetracaine on the ionic current in enzymatically dissociated single guinea-pig ventricular cells was studied using a two micro-electrode voltage-clamp technique. The myocytes were pre-incubated with Cs+ and the experiments were performed at room temperature in order to reduce the contribution of the delayed outward current. Tetracaine decreased the maximum rate of rise of the action potential with a dissociation constant (KD) strongly dependent on the holding potential (0.77 microM at -80 mV, and 6.2 microM at -95 mV). Application of 20 microM-tetracaine resulted in about a 50% reduction of the inwardly rectifying K+ current, while ten times higher concentrations were required to suppress the delayed K+ current. The inactivation time course of the Ca2+ current could be fitted with two exponentials, with time constants tau f = 15 ms and tau s = 150 ms at around 0 mV. Tetracaine decreased the amplitude of the Ca2+ current and speeded its decay. This effect was found to be primarily due to a marked inhibition of the amplitude of the slowly inactivating component (apparent KD = 80 microM, nH = 2). The drug had little effect on the time constants of the two components of Ca2+ channel inactivation. When Sr2+ or Ba2+ were the charge carriers, inactivation of the Ca2+ channel was again fitted with a fast and a slow exponential. In addition, a maintained (or very slowly inactivating) component was present. Tetracaine not only suppressed the amplitudes of the slowly inactivating and the maintained components, but also decreased the time constant of the slowly inactivating component. The results are consistent with a direct effect of tetracaine on the high threshold Ca2+ channel and do not support indirect effects of the drug secondary to suppression of Ca2+ release from internal stores.