Delayed rectifier potassium channels in canine and porcine airway smooth muscle cells

Abstract

1. In order to define the ion channels underlying the inactivating, calcium-insensitive current in airway smooth muscle cells, unitary potassium currents were recorded from canine and porcine trachealis cells, and compared with macroscopic currents. On-cell and inside-out single-channel currents were compared with whole-cell recordings made in dialysed cells. 2. Depolarizing voltage steps evoked outward unitary currents. In addition to a large conductance, calcium-activated potassium channel (KCa), a lower conductance potassium channel was identified. This channel has a conductance of 12.7 pS (on-cell; 1 mM-K+ in the pipette). 3. The lower conductance channel (Kdr) was not sensitive to cytosolic Ca2+ concentration and unitary current openings occurred following a delay after the voltage step. The time course of activation of the current composed of averaged single-channel events was very similar to that of the whole-cell, delayed rectifier potassium current (IdK), recorded under conditions of low intracellular calcium (Kotlikoff, 1990). 4. Kdr channels also inactivated with kinetics similar to those of the macroscopic current. Averaged single-channel records revealed a current that inactivated with kinetics that could be described by two exponentials (tau 1 = 0.14 s, tau 2 = 1.1 s; at 5 mV). These values corresponded well with previously determined values for time-dependent inactivation of IdK. Inactivation of Kdr channels was markedly voltage dependent, and was well fitted by a Boltzmann equation with V50 = -53 mV; this was similar to measurements of the macroscopic current, although the V50 value was shifted to more positive potentials in whole-cell measurements. When only the inactivating component of the macroscopic current was considered, the voltage dependence of inactivation of the single-channel current and macroscopic current were quite similar. 5. Single-channel kinetics indicated that Kdr channels occupy one open and two closed states. The mean open time was 1.7 ms. Inactivation results in a prominent increase in the long closed time, with little effect on the mean open time or short closed time. 6. The Kdr channel was not blocked by tetraethylammonium (TEA; 1 mM), charybdotoxin (ChTX; 100 nM) or glibenclamide (20 microM), but was blocked by 4-aminopyridine (4-AP; 1 mM). Similarly, 4-AP blocked the inactivating component of the macroscopic current, but a non-inactivating current remained. KCa currents were blocked by TEA (0.5-1 mM) and charybdotoxin (40 nM), but were insensitive to to 4-AP (1 mM) and glibenclamide (20 microM).(ABSTRACT TRUNCATED AT 400 WORDS)