Potential-dependent calcium inward current in a single isolated smooth muscle cell of the guinea-pig taenia caeci

Abstract

A single glass micropipette voltage-clamp technique was used to study a potential-dependent calcium inward current in isolated smooth muscle cells of the guinea-pig taenia caeci. Experiments were performed at 22-24 degrees C. With potassium as the main cation in the pipette solution, a transient inward current appeared in response to a depolarizing pulse, followed by an outward current. The replacement of potassium ions by caesium ions and TEA (tetraethyl-ammonium) in the pipette solution resulted in an effective suppression of potassium outward current permitting a study of the calcium current solely. The calcium inward current was blocked by 5 mM-cobalt and 5 X 10(-6) M-verapamil. Activation of the calcium current occurred at a membrane potential of between -35 and -25 mV. The calcium current was maximal in the potential range +10 to +20 mV and did not reverse even at +60 mV. Inactivation of the calcium current had a complex nature. It did not inactivate completely even during depolarizations lasting many seconds. During the first 400 ms the decay of the calcium current followed a time course described by two exponentials. The fast time constant of decay was in the range of 40 to 53 ms (n = 3) and the slow time constant was approximately 10-fold greater (at 0 mV). The fast time constant did not depend on the membrane potential while the slow time constant decreased with depolarization. Availability of the calcium current was estimated in double-pulse experiments. It had a U-shaped dependence on the conditioning potential; maximal inactivation was observed at potentials corresponding to the maximal calcium current. It was suggested that a component of inactivation was dependent on the calcium current which flowed. Calculations of calcium entry at various depolarizations showed that large amounts of calcium ions enter the cell. Also, it was suggested that calcium ions are effectively bound within the smooth muscle cell.