Noma A, Shibasaki T
J Physiol. 1985 Jun;363:463-80. doi: 10.1113/jphysiol.1985.sp015722.
The question whether activation of the ATP-regulated K channel is responsible for macroscopic anoxia-induced outward currents was examined in ventricular cells isolated enzymatically from guinea-pig heart. Gigaseal patch-clamp electrodes were used for a whole-cell voltage clamp. Membrane currents were compared in the same cell while the cell interior was dialysed by perfusing the electrode with different solutions. When the cell was dialysed with various ATP-deficient (less than or equal to 2 mM) internal solutions, the Ca current decreased in a dose-dependent manner to less than 10% of control at 0.5 mM-ATP. A slight (ca. 25%) decrease of the slope conductance for hyperpolarizing current was observed. When a delayed rectification on depolarization followed by a marked outward current tail on repolarization was present under control conditions, this time-dependent outward current was also depressed. An increase in a time-independent outward current was observed accompanied by marked current fluctuations. The outward current showed a reversal potential near the K equilibrium potential, inward rectification, and no relaxation on voltage jumps. The power density spectrum of the current fluctuations showed a pattern similar to the spectrum calculated from the single-channel currents of ATP-regulated K channels. The amplitude of the single-channel current, estimated from the fluctuations, was almost equal to that of the single-channel current. The total number of channels within one cell was estimated as 2000-3000. It is concluded that the ATP-regulated K channels are responsible for the increase in the outward current and the shortening of the action potential duration under various anoxic conditions.
采用酶解法从豚鼠心脏分离出心室细胞,研究了ATP调节的钾通道激活是否是宏观缺氧诱导外向电流的原因。使用千兆封接膜片钳电极进行全细胞电压钳制。在同一细胞中,通过用不同溶液灌注电极来透析细胞内部,比较膜电流。当用各种ATP缺乏(小于或等于2 mM)的内部溶液透析细胞时,Ca电流以剂量依赖的方式降低,在0.5 mM-ATP时降至对照的10%以下。观察到超极化电流的斜率电导略有(约25%)下降。在对照条件下,当去极化时出现延迟整流,复极化时出现明显的外向电流尾时,这种时间依赖性外向电流也受到抑制。观察到非时间依赖性外向电流增加,并伴有明显的电流波动。外向电流在钾平衡电位附近显示出反转电位,内向整流,电压阶跃时无松弛。电流波动的功率密度谱显示出与从ATP调节的钾通道单通道电流计算出的谱相似的模式。根据波动估计的单通道电流幅度几乎与单通道电流幅度相等。估计一个细胞内通道总数为2000-3000。结论是,ATP调节的钾通道负责在各种缺氧条件下外向电流的增加和动作电位持续时间的缩短。