Inhibition of delayed rectifier K(+)-current by levcromakalim in single intestinal smooth muscle cells: effects of cations and dependence on K(+)-flux.
1. Whole-cell voltage-clamp recordings were made from single smooth muscle cells isolated from the longitudinal layer of the guinea-pig small intestine. 2. Levcromakalim ((-)Ckm) inhibited delayed rectifier K-current (IK(DR)) and induced a voltage-independent K-current (IK(-Ckm)). Both effects were inhibited similarly by glibenclamide. In some cells, however, IK(-Ckm) could be induced without any effect on IK(DR). 3. Ba2+ caused a voltage-dependent block of IK(-Ckm). The IC50 was 0.2 mM at -40 mV (6 cells), but at 0 mV 2 mM Ba2+ caused only a 26 +/- 7% inhibition (n = 5). Ba2+ had much less effect on IK(DR), 2 mM Ba2+ having no inhibitory effect on current elicited by depolarization to -30 mV (n = 6) or 0 mV (n = 5). 4. Low concentrations of Zn2+ blocked IK(-Ckm) while having little effect on IK(DR). Zn2+ (40 microM) caused a 77 +/- 1% reduction of IK(-Ckm) at -30 mV (n = 4) but IK(DR) was inhibited by only 10 +/- 3% at the same voltage (n = 4). 5. Inward current amplitudes were compared in 135 mM Rb+ and 135 mM K+ bath solutions. (-)Ckm-activated Rb(+)-current was only 4% of the K(+)-current, whereas delayed rectifier Rb(+)-current was larger than K(+)-current. 6. (-)Ckm did not inhibit IK(DR) if IK(-Ckm) was blocked. In the presence of 2 mM Ba2+ or 135 mM Rb+, (-)Ckm did not induce current nor did it inhibit the delayed rectifier. When [Rb+]o was 25 mM and [K+]J was 130 mM, (-)Ckm elicited outward current and inhibited outward delayed rectifier current (at voltages positive of the reversal potential) but it did not elicit inward current or inhibit inward delayed rectifier current (at voltages negative of the reversal potential).7. These experiments indicate that (-)Ckm-activated K channels are more sensitive to inhibition by Ba2+and Zn2+ and pass inward Rb+ current less well than delayed rectifier K channels. They also suggest that (-)Ckm does not modulate delayed rectifier K channels directly or via an intermediate protein but that the inhibitory effect of (-)Ckm on IK(DR) arises as a consequence of K+-flux through (-)Ckm activated K channels.
McHugh
D
D
Department of Pharmacology, University of Leeds.
Beech
D J
DJ
D000818:Animals; D001464:Barium / Q000494:pharmacology; D001578:Benzopyrans / Q000494:pharmacology*; D002412:Cations / Q000494:pharmacology*; D006224:Cricetinae; D019806:Cromakalim; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007421:Intestine, Small / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism*; D015221:Potassium Channels / Q000187:drug effects*; D011758:Pyrroles / Q000494:pharmacology*; D012413:Rubidium / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology
D001578:Benzopyrans; D002412:Cations; D015221:Potassium Channels; D011758:Pyrroles; D019806:Cromakalim; D001464:Barium; D015032:Zinc; D012413:Rubidium; D011188:Potassium