Calcium currents in isolated canine airway smooth muscle cells

Abstract

Canine tracheal smooth muscle cells were enzymatically dissociated, and individual myocytes were voltage clamped through use of the whole cell, patch-clamp method. Cells dialyzed with solutions high in potassium and bathed in physiological saline demonstrated brief inward currents, followed by large outward currents that inactivated very slowly. When outward currents were blocked, a voltage-activated inward current was observed that activated with depolarizations to voltages positive to -45 mV, with an apparent reversal potential greater than 110 mV, and a peak current at 15 mV. This current was identified as a calcium current on the basis of 1) its presence under conditions in which calcium was the only permeant cation, 2) the lack of a blocking effect of 2 microM tetrodotoxin, and 3) block of the current by Mn2+, Cd2+, and CO2+. Increases in external calcium concentration from 2 to 20 mM resulted in an increase in current amplitude and a shift of voltage activation toward more positive potentials. The current displayed a rapid inactivation phase with a time constant of 16-52 ms, which was well fit by a single exponential. Steady-state inactivation of the calcium current was sigmoidal, with a voltage of half inactivation of -21 mV in 20 mM Ca2+. The principle component of the calcium current was further identified as a transient current on the basis of its rapid inactivation, current-voltage characteristics, and relative insensitivity to dihydropyridine calcium channel blocking agents.