Shapiro M S, DeCoursey T E
Department of Physiology, Rush Presbyterian St. Luke's Medical Center, Chicago, Illinois 60612.
J Gen Physiol. 1991 Jun;97(6):1227-50. doi: 10.1085/jgp.97.6.1227.
Type l voltage-gated K+ channels in murine lymphocytes were studied under voltage clamp in cell-attached patches and in the whole-cell configuration. The kinetics of activation of whole-cell currents during depolarizing pulses could be fit by a single exponential after an initial delay. Deactivation upon repolarization of both macroscopic and microscopic currents was mono-exponential, except in Rb-Ringer or Cs-Ringer solution in which tail currents often displayed "hooks," wherein the current first increased or remained constant before decaying. In some cells type l currents were contaminated by a small component due to type n K+ channels, which deactivate approximately 10 times slower than type l channels. Both macroscopic and single channel currents could be dissected either kinetically or pharmacologically into these two K+ channel types. The ionic selectivity and conductance of type l channels were studied by varying the internal and external permeant ion. With 160 mM K+ in the cell, the relative permeability calculated from the reversal potential with the Goldman-Hodgkin-Katz equation was K+ (identical to 1.0) greater than Rb+ (0.76) greater than NH4+ = Cs+ (0.12) much greater than Na+ (less than 0.004). Measured 30 mV negative to the reversal potential, the relative conductance sequence was quite different: NH4+ (1.5) greater than K+ (identical to 1.0) greater than Rb+ (0.5) greater than Cs+ (0.06) much greater than Na+, Li+, TMA+ (unmeasurable). Single channel current rectification resembled that of the whole-cell instantaneous I-V relation. Anomalous mole-fraction dependence of the relative permeability PNH4/PK was observed in NH4(+)-K+ mixtures, indicating that the type l K+ channel is a multi-ion pore. Compared with other K+ channels, lymphocyte type l K+ channels are most similar to "g12" channels in myelinated nerve.
在细胞贴附式膜片钳和全细胞模式下,利用电压钳对小鼠淋巴细胞中的I型电压门控钾通道进行了研究。去极化脉冲期间全细胞电流的激活动力学在初始延迟后可用单指数函数拟合。宏观电流和微观电流复极化时的失活均为单指数形式,但在铷林格氏液或铯林格氏液中,尾电流常呈现“钩状”,即电流在衰减前先增大或保持恒定。在一些细胞中,I型电流被一小部分由于II型钾通道产生的电流污染,其失活速度比I型通道慢约10倍。宏观电流和单通道电流在动力学或药理学上都可分为这两种钾通道类型。通过改变细胞内、外通透离子研究了I型通道的离子选择性和电导。当细胞内钾离子浓度为160 mM时,根据戈德曼-霍奇金- Katz方程由反转电位计算出的相对通透性为:钾离子(等于1.0)大于铷离子(0.76)大于铵离子 = 铯离子(0.12)远大于钠离子(小于0.004)。在比反转电位负30 mV处测量时,相对电导顺序有很大不同:铵离子(1.5)大于钾离子(等于1.0)大于铷离子(0.5)大于铯离子(0.06)远大于钠离子、锂离子、四甲基铵离子(无法测量)。单通道电流整流类似于全细胞瞬时电流-电压关系。在铵离子-钾离子混合物中观察到相对通透性PNH4/PK的异常摩尔分数依赖性,表明I型钾通道是一种多离子孔道。与其他钾通道相比,淋巴细胞I型钾通道与有髓神经中的“g12”通道最为相似。