Single channel and whole-cell K-currents evoked by levcromakalim in smooth muscle cells from the rabbit portal vein

Abstract

1. Single channel and whole-cell current recordings were made from single smooth muscle cells isolated from the rabbit portal vein. 2. Application of 10 microM levcromakalim ((-)-Ckm) to single cells held with pipettes containing 1 mM GDP induced a K-current (IK(Ckm)) which occurred in addition to the current caused by GDP alone (IK(GDP)) and averaged 135 pA at -37 mV. We have investigated whether the same K channels underlie the GDP- and Ckm-induced K-currents. 3. If 1 mM GDP was in the pipette but Mg ions were omitted the effect of GDP was absent and IK(Ckm) averaged only 10 pA, suggesting that the action of (-)-Ckm was Mg-dependent. 4. Intracellular ATP was not observed to have much effect on IK(-Ckm). Loading of cells with 10 mM ATP from the recording pipette had no significant effect and flash photolysis of caged-ATP loaded into cells from the pipette, estimated to release about 1 mM free ATP, also had no effect on IK(-Ckm). 5. Bath-applied glibenclamide inhibited IK(-Ckm) with an IC50 of 200 nM, a value 8 times higher than that found for inhibition of IK(GDP). The delayed rectifier K-current (IK(DR)) was also inhibited by glibenclamide but at higher concentrations (IC50 100 microM). Bath-applied tetraethylammonium ions (TEA) inhibited IK(-Ckm) and IK(GDP) to the same extent (IC50 about 7 mM). 6. In inside-out patch recordings (- )-Ckm (10 microM) applied to the intracellular surface of the membrane potentiated the opening of K channels already stimulated by I mM GDP and all of the channel activity was abolished by 10 microM glibenclamide. The unitary conductance of the channels was 24lpS in a 60 mM: 130 mM K-gradient.7. We suggest that (-)-Ckm may hyperpolarize and relax smooth muscle cells by opening KNDP, a class of small conductance K channels that are related to the ATP-sensitive K channels seen in other tissues.