A large conductance [Ca(2+)](i)-independent K(+) channel expressed in HaCaT keratinocytes

Patch-clamp recordings were carried out in the inside-out configuration in human keratinocytes of the cell line HaCaT. Patch pipettes were filled with 150 mM KCl, 1 mM CaCl(2) and 10 mM HEPES. In symmetrical KCl solutions, single channel currents from a large conductance channel (about 170 pS) were measured. Replacement of 120 mM KCl by K-aspartate had only a minor influence on the single channel conductance and on the reversal potential. In intracellular solution in which K(+) has been replaced by Na(+) or NMDG(+), the reversal potential shifted to > + 40 mV indicating K(+) as the main charge carrier. The channels were neither dependent on intracellular Ca(2+) (between 0.8 nM and 10 micro M), ATP (at 0 and 1 mM) nor Mg(2+) (at 0 and 0.5 mM). The mean current showed an outward rectification that can be mainly attributed to the voltage dependence of the open probability. The channels displayed bursting kinetics with a mean open time of about 2 ms and closed times of about 0.2, 2 and 20 ms. The mean open probability was usually low (0.05) but increased occasionally (0.6) mainly due to a lower probability of long closings. We conclude that these K(+) channels contribute to the resting potential of human keratinocytes which may control the Ca(2+) influx and thereby their proliferation and differentiation.