Saunders H H, Farley J M
Department of Pharmacology and Toxicology, University of Mississippi Medical Center, Jackson.
J Pharmacol Exp Ther. 1992 Mar;260(3):1038-44.
The pharmacological properties of large conductance Ca(++)-activated K+ channels and spontaneous transient outward currents (STOCs) in swine tracheal smooth muscle (TSM) were studied using enzymatically dissociated single cells and patch clamp techniques. Recording from inside-out patches showed that extracellular tetraethylammonium caused a concentration-dependent decrease in single channel current amplitude. Acetylcholine (ACh; 10(-8) M) induced repetitive outward current transients in cell-attached patches due to simultaneous opening of many large conductance Ca(++)-activated K+ channels. Whole-cell recordings revealed the presence of STOCs in TSM membrane that were inhibited by tetraethylammonium at concentrations similar to those which inhibited the large conductance Ca(++)-activated K+ channels. Caffeine (5 mM) stimulated and then inhibited STOCs, suggesting that they are induced by the release of calcium from internal stores. ACh (1 microM) initially increased STOC frequency followed by an inhibition of STOCs. The effects of ACh occurred in control solution (2 mM Ca++) or when calcium influx was eliminated (0 Ca++, 2 mM Mn++). ACh also induced an inward current at negative membrane potentials associated with an increase in conductance. We conclude that inhibition of STOCs (i.e., potassium channel activity) and induction of an inward cation current by ACh may give rise to the depolarization observed in the presence of ACh and be important in the development of ACh-induced contractions in TSM.
运用酶解分离的单细胞和膜片钳技术,研究了猪气管平滑肌(TSM)中大电导钙激活钾通道的药理学特性和自发性瞬时外向电流(STOCs)。从外向内膜片记录显示,细胞外四乙铵导致单通道电流幅度呈浓度依赖性降低。乙酰胆碱(ACh;10⁻⁸ M)在细胞贴附式膜片中诱导重复性外向电流瞬变,这是由于许多大电导钙激活钾通道同时开放所致。全细胞记录显示TSM膜中存在STOCs,其在与抑制大电导钙激活钾通道相似的浓度下被四乙铵抑制。咖啡因(5 mM)先刺激然后抑制STOCs,表明它们是由细胞内钙库释放钙所诱导。ACh(1 μM)最初增加STOC频率,随后抑制STOCs。ACh的这些作用在对照溶液(2 mM Ca²⁺)中或消除钙内流(0 Ca²⁺,2 mM Mn²⁺)时均会发生。ACh在负膜电位时还诱导与电导增加相关的内向电流。我们得出结论,ACh对STOCs(即钾通道活性)的抑制以及对内向阳离子电流的诱导可能导致在ACh存在时观察到的去极化,并在ACh诱导的TSM收缩的发生中起重要作用。