Blatz A L, Magleby K L
Biophys J. 1985 Jan;47(1):119-23. doi: 10.1016/S0006-3495(85)83884-4.
The patch-clamp technique was used to characterize channels that could contribute to the resting Cl-conductance in the surface membrane of cultured rat skeletal muscle. Two Cl- -selective channels, in addition to the Cl- -selective channel of large conductance described previously (Blatz and Magleby, 1983), were observed. One of these channels had fast kinetics and a conductance of 45 +/- 1.8 pS (SE) in symmetrical 100 mM KCl. The other had slow kinetics and a conductance of 61 +/- 2.4 pS. The channel with fast kinetics typically closed within 1 ms after opening and flickered between the open and shut states. The channel with slow kinetics typically closed within 10 ms after opening and displayed less flickering. Both channels were active in excised patches of membrane held at potentials similar to resting membrane potentials in intact cells, and both were open a greater percentage of time with depolarization. Under conditions of high ion concentrations, both channels exhibited nonideal selectivity for Cl- over K+ with the permeability ratio PK/PCl of 0.15-0.2. Additional experiments on the fast Cl- channel indicated that its activity decreased with lowered pHi and that SO2-4 and CH3SO-4 were ineffective charge carriers. These findings, plus the observation that the fast Cl- channel was also active in membrane patches on intact cells, suggest that the fast Cl- channel provides a molecular basis for at least some of the resting Cl- conductance. The extent to which the slow Cl- channel contributes is less clear as it was typically active only after excised patches of membrane had been exposed to high concentrations of KCl at the inner membrane surface.
采用膜片钳技术对可能参与培养的大鼠骨骼肌表面膜静息氯电导的通道进行了特性分析。除了先前描述的大电导氯选择性通道(Blatz和Magleby,1983)外,还观察到另外两个氯选择性通道。其中一个通道动力学较快,在对称的100 mM KCl中电导为45±1.8 pS(标准误)。另一个通道动力学较慢,电导为61±2.4 pS。动力学较快的通道通常在开放后1毫秒内关闭,并在开放和关闭状态之间闪烁。动力学较慢的通道通常在开放后10毫秒内关闭,闪烁较少。在与完整细胞静息膜电位相似的电位下,两个通道在切除的膜片中均有活性,并且随着去极化,它们开放的时间百分比更高。在高离子浓度条件下(条件下),两个通道对Cl-相对于K+均表现出非理想选择性,通透率PK/PCl为0.15 - 0.2。对快速氯通道的进一步实验表明,其活性随细胞内pH值降低而降低,并且SO42-和CH3SO4-是无效的电荷载体。这些发现,加上快速氯通道在完整细胞膜片上也有活性的观察结果,表明快速氯通道至少为部分静息氯电导提供了分子基础。慢速氯通道的贡献程度尚不清楚,因为它通常仅在切除的膜片内膜表面暴露于高浓度KCl后才具有活性。