Robertson B E, Bonev A D, Nelson M T
Department of Pharmacology, University of Vermont, Colchester 05446, USA.
Am J Physiol. 1996 Aug;271(2 Pt 2):H696-705. doi: 10.1152/ajpheart.1996.271.2.H696.
Inward rectifier K+ channels have been implicated in the control of membrane potential and external K(+)-induced dilations of small coronary arteries. To identify and characterize inward rectifier K+ currents in coronary artery smooth muscle, whole cell K+ currents in smooth muscle cells enzymatically isolated from rat coronary (septal) arteries (diameters, 100-150 microns) were measured in the conventional and perforated configurations of the patch-clamp technique. Ba(2+)-sensitive, whole cell K+ current-voltage relationships exhibited inward rectification. Blockers of Ca(2+)-activated K+ channels (1 mM tetraethylammonium ion), ATP-sensitive K+ channels (10 microM glibenclamide), and voltage-dependent K+ channels (1 mM 4-aminopyridine) in smooth muscle did not affect inward rectifier K+ currents. The nonselective K+ channel inhibitor phencyclidine (100 microM) reduced inward rectifier K+ currents by approximately 50%. External Ba2+ reduced inward currents, with membrane potential hyperpolarization increasing inhibition. The half-inhibition constant for Ba2+ was 2.1 microM at -60 mV, decreasing e-fold for a 25-mV hyperpolarization. External Cs+ also blocked inward rectifier K+ currents, with the half-inhibition constant for Cs+ of 2.9 mM at -60 mV. External Ca2+ and Mg2+ reduced inward rectifier K+ currents. At -60 mV, Ca2+ and Mg2+ (1 mM) reduced inward currents by 33 and 21%, respectively. Inward rectification was not affected by dialysis of the cell's interior with a nominally Ca(2+)- and Mg(2+)-free solution. These findings indicate that inward rectifier K+ channels exist in coronary artery smooth muscle and that Ba2+ may be a useful probe for the functional role of inward rectifier K+ channels in coronary arteries.
内向整流钾通道与膜电位的调控以及外源性钾离子诱导的小冠状动脉扩张有关。为了鉴定和表征冠状动脉平滑肌中的内向整流钾电流,采用膜片钳技术的常规配置和穿孔配置,测量了从大鼠冠状动脉(间隔支)酶分离的平滑肌细胞(直径100 - 150微米)中的全细胞钾电流。钡离子敏感的全细胞钾电流 - 电压关系呈现内向整流特性。平滑肌中钙激活钾通道阻滞剂(1 mM四乙铵离子)、ATP敏感性钾通道阻滞剂(10 microM格列本脲)和电压依赖性钾通道阻滞剂(1 mM 4 - 氨基吡啶)均不影响内向整流钾电流。非选择性钾通道抑制剂苯环己哌啶(100 microM)使内向整流钾电流降低约50%。外源性钡离子降低内向电流,膜电位超极化时抑制作用增强。在 - 60 mV时,钡离子的半抑制常数为2.1 microM,膜电位超极化25 mV时抑制常数降低10倍。外源性铯离子也阻断内向整流钾电流,在 - 60 mV时铯离子的半抑制常数为2.9 mM。外源性钙离子和镁离子降低内向整流钾电流。在 - 60 mV时,钙离子和镁离子(1 mM)分别使内向电流降低33%和21%。用名义上无钙和无镁的溶液透析细胞内部,内向整流特性不受影响。这些发现表明冠状动脉平滑肌中存在内向整流钾通道,钡离子可能是研究内向整流钾通道在冠状动脉中功能作用的有用探针。