ABSTRACT
The properties of the outward Ca(2+)-dependent K+ current (KCa) were investigated in single smooth muscle cells (SMCs) isolated from epididymal part of the rat vas deferens (RVD) using amphotericin B perforated patch-clamp technique. The complex kinetic of the net outward current elicited by positive voltage steps from -80 mV to +40 mV suggested the presence of several components of this current. KCa current was separated from the net outward current by removal of Ca2+ from the external solution. KCa was characterized by slow kinetics of current activation and decay. Mycotoxin paxilline, the selective blocker of the large conductance KCa channels, inhibited KCa current in a dose-dependent manner. At the concentration of 70 nM paxilline evoked 50% inhibition of KCa and at 1 mkM complete suppression of KCa current was achieved. The blocking effect of low concentrations of a nonselective KCa channels inhibitor tetraethylammonium (TEA) was compared to that of paxilline. The external application of 0.3 mM TEA inhibited KCa current similarly to 1 mkM of paxilline. Finally, we studied the effect of paxilline on the resting membrane potential ofRVD SMCs. Paxilline (1 mkM) did not affect the membrane potential of SMCs with the resting potential in the range of -60 to -40 mV. However, at potentials more positive than -40 mV application of paxilline significantly (up to 15 mV) depolarized the membrane of SMCs. These results suggest that the large conductance KCa channels in RVD SMCs do not contribute to the resting membrane potential but could serve as a hyperpolarizing mechanism at the significant membrane depolarizations.