Kimitsuki T, Mitsuiye T, Noma A
Department of Physiology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Pflugers Arch. 1990 Jul;416(5):493-500. doi: 10.1007/BF00382681.
Single Na+ channel currents were recorded from guinea-pig ventricular cells in cell-attached patches. The ensemble average current (I) of multi-channel recordings was used to calculate the variance (sigma 2) of current fluctuations around the mean in individual current recordings. The relationship between sigma 2/I and I was linear and allowed estimation of the number of functional channels in the patch of membrane. The unitary amplitude of channel current obtained from the relation sigma 2/I-I was in agreement with that obtained directly by measuring the original records. The number of channels determined at different depolarizing pulses was almost constant in a given patch. The value was nearly equal to that of the maximum current, measured at high depolarizing potentials when most channels are open, divided by the unitary current. The open probability of the channels at the peak time of mean current was calculated based on the estimated number of channels. It increased with increasing depolarization and saturated at about 0.6 at test potentials above -20 mV. The inactivation time-course of the mean current was fitted by a sum of two exponentials. The current amplitude extrapolated to time zero was much larger than the current which could be generated by all channels. This indicates that the inactivation of the Na+ channel develops with delay after the onset of depolarization. The finding is in agreement with a model in which the inactivation rate is accelerated with activation of the Na+ channel.
在细胞贴附式膜片钳记录豚鼠心室肌细胞的单个钠通道电流。多通道记录的总体平均电流(I)用于计算单个电流记录中围绕平均值的电流波动方差(σ2)。σ2/I与I之间的关系呈线性,可据此估计膜片上功能通道的数量。由σ2/I-I关系得出的通道电流单位幅值与直接测量原始记录所得结果一致。在给定的膜片中,不同去极化脉冲下测定的通道数量几乎恒定。该值几乎等于在高去极化电位(此时大多数通道开放)下测得的最大电流除以单位电流。根据估计的通道数量计算平均电流峰值时刻通道的开放概率。其随去极化增强而增加,在测试电位高于-20 mV时约为0.6达到饱和。平均电流的失活时间进程用两个指数之和拟合。外推至时间零点的电流幅值远大于所有通道可能产生的电流。这表明钠通道的失活在去极化开始后延迟出现。该发现与钠通道激活加速失活速率的模型一致。