Kakei M, Noma A
J Physiol. 1984 Jul;352:265-84. doi: 10.1113/jphysiol.1984.sp015290.
The patch-clamp method was applied to single atrioventricular (a.v.) node cells of the rabbit heart to study the characteristics of the K+ channel. When the electrode contained 5.4 mM-K+, depolarizations of the cell-attached patch membrane induced outward single channel currents characterized by burst-like openings; the open-state probability increased from 0.005-0.01 at -40 mV to 0.07-0.1 at +20 mV of membrane potential. The reversal potentials of the current at K+ concentrations of 5.4, 20 and 130 mM in the electrode agreed with those given by the Nernst equation, indicating that this channel is selective for K+ ions. The slope conductance of the channel decreased beyond 60-90 mV positive to the reversal potential (inward-going rectification). The conductance near the reversal potential increased with increasing K+ concentrations on either side of the membrane: from 31-32 pS at 5.4 mM-K+ to 41-42 pS at 20 mM-K+ on the outside, and from 19 pS at 90 mM-K+ to 29.3 pS at 130 mM-K+ on the inside. Superfusion of the cell with 5.4 mM-CN-, glucose-free Tyrode markedly increased the number of channel openings in the cell-attached patch. In the inside-out patch, application of 1 mM-adenosine-5'-triphosphate (ATP) at the inner surface of the patch membrane blocked reversibly the channel activity, while 1 mM-adenosine-5'-diphosphate (ADP) failed to block it. The conductance and kinetics of the channel were not modified by increasing the Ca2+ concentration from 10(-8) M to 5 X 10(-6) M on the inner side of the membrane, while a further increase in Ca2+ to 10(-4) M decreased the open-state probability. The probability density for the open time fitted well with an exponential distribution (time constant of 5.4 ms at 60 mV positive to the resting potential), while that for the closed time was separated into a fast and a slow component (time constants of 4.0 and 132.0 ms, respectively). The time constant of the slow component decreased significantly with depolarization in some preparations. However, neither the time constant of the fast component of the closed-time histogram nor that of the open-time histogram was voltage-dependent.
采用膜片钳技术研究兔心脏单个房室(a.v.)结细胞的钾通道特性。当电极内含有5.4 mM - K⁺时,细胞贴附式膜片膜的去极化诱导出外向单通道电流,其特征为爆发式开放;开放概率从膜电位 - 40 mV时的0.005 - 0.01增加到 + 20 mV时的0.07 - 0.1。电极内K⁺浓度为5.4、20和130 mM时电流的反转电位与能斯特方程计算值相符,表明该通道对K⁺离子具有选择性。通道的斜率电导在超过反转电位正向60 - 90 mV时降低(内向整流)。反转电位附近的电导随膜两侧K⁺浓度增加而增大:外侧5.4 mM - K⁺时为31 - 32 pS,20 mM - K⁺时为41 - 42 pS;内侧90 mM - K⁺时为19 pS,130 mM - K⁺时为29.3 pS。用不含葡萄糖的5.4 mM - CN⁻ - Tyrode液灌流细胞,显著增加了细胞贴附式膜片中通道的开放次数。在膜内面向外的膜片中,在膜片内表面施加1 mM - 腺苷 - 5'-三磷酸(ATP)可可逆性阻断通道活性,而1 mM - 腺苷 - 5'-二磷酸(ADP)则无此作用。膜内侧Ca²⁺浓度从10⁻⁸ M增加到5×10⁻⁶ M时,通道的电导和动力学未改变,但Ca²⁺进一步增加到10⁻⁴ M时,开放概率降低。开放时间的概率密度很好地符合指数分布(相对于静息电位正向60 mV时的时间常数为5.4 ms),而关闭时间的概率密度分为快速和慢速成分(时间常数分别为4.0和132.0 ms)。在某些标本中,慢速成分的时间常数随去极化显著降低。然而,关闭时间直方图的快速成分时间常数和开放时间直方图的时间常数均与电压无关。