Takanashi H, Sawanobori T, Kamisaka K, Maezawa H, Hiraoka M
Third Department of Internal Medicine, Tokyo Medical and Dental University, Japan.
J Cell Physiol. 1994 Dec;161(3):537-43. doi: 10.1002/jcp.1041610317.
The patch-clamp technique of cell-attached and inside-out configurations was used to study the single potassium channels in isolated guinea pig hepatocytes. The single potassium channels in isolated guinea pig hepatocytes were recorded at different K+ concentrations. A linear single-channel current-voltage relationship was obtained at the voltage range of -80 to -20 mV with slope conductance of 70 +/- 6 pS (n = 10). Under symmetrical high K+ concentration of 148 mM in the cell-attached patch membrane, the I-V curve exhibited a mild inward rectification at potentials positive to + 20 mV. The values of reversal potential was +5 +/- 2 mV (n = 10). When the external potassium concentration ([K+]o) was decreased to 74 mM and 20 mM, the slope conductance was decreased to 48 +/- 2 pS (n = 4) and 24 +/- 3 pS (n = 3), respectively. The reversal potential was changed by 58 mV for a tenfold change in [K+]o, indicating that this channel was highly selective for K+. Open probabilities (Po) of the channel were 73-93% without apparent voltage dependence. The distributions of open time of the channels were fitted to two exponentials, while those of closed time were fitted to three exponentials, exhibiting no voltage dependence. The success rate of K+ channel activity to be recorded was 28% at room temperature, and there were no increases in the success rate nor in the channel opening probabilities at a temperature of 34-36 degrees C. Po in inside-out patches was not changed by application of 1 microM Ca2+ nor 1 mM Mg2+ to the internal side of patch membranes. It is concluded that a novel type of the K+ channels in guinea pig hepatocytes had different properties of slope conductance, channel kinetics, and sensitivity to [Ca2+]i, from those in other species.
采用细胞贴附式和内面向外式膜片钳技术研究分离的豚鼠肝细胞中的单个钾通道。在不同的K⁺浓度下记录分离的豚鼠肝细胞中的单个钾通道。在-80至-20 mV的电压范围内获得线性单通道电流-电压关系,斜率电导为70±6 pS(n = 10)。在细胞贴附式膜片的膜内对称高K⁺浓度为148 mM时,I-V曲线在正于+20 mV的电位处表现出轻度内向整流。反转电位值为+5±2 mV(n = 10)。当外部钾浓度([K⁺]o)降至74 mM和20 mM时,斜率电导分别降至48±2 pS(n = 4)和24±3 pS(n = 3)。[K⁺]o每变化10倍,反转电位变化58 mV,表明该通道对K⁺具有高度选择性。通道的开放概率(Po)为73 - 93%,无明显电压依赖性。通道开放时间分布拟合为两个指数函数,而关闭时间分布拟合为三个指数函数,均无电压依赖性。在室温下记录K⁺通道活性的成功率为28%,在34 - 36℃时成功率和通道开放概率均未增加。向膜片内侧施加1 μM Ca²⁺或1 mM Mg²⁺不会改变内面向外式膜片中的Po。结论是豚鼠肝细胞中的一种新型K⁺通道在斜率电导、通道动力学和对[Ca²⁺]i的敏感性方面与其他物种的不同。