Barish M E, Baud C
J Physiol. 1984 Jul;352:243-63. doi: 10.1113/jphysiol.1984.sp015289.
Membrane currents in the immature oocyte of the urodele amphibian Ambystoma were studied using the two-micro-electrode voltage-clamp technique. A current carried by H ions (IH) constituted the major portion of outward current activated by depolarizations from the resting voltage (about -60 mV). Net inward current was not observed at this developmental stage. The reversal potential for IH measured from tail currents obtained in two step voltage-clamp experiments shifted by 54 mV per unit change in external pH between pH 6.9 and 8.4. The reversal potential at pH 7.4 was not affected by changes in external K or Cl concentrations. A small change in reversal potential was observed with removal of external Na. The amplitude of IH was not affected by removal of external Ca (Mg or Sr substitution). Ca ionophore A23187 shifted the current-voltage relation towards negative voltages. Activation of IH did not appear to depend on Ca influx. The instantaneous current-voltage relation for IH, measured from tail currents at approximately equal internal and external H ion concentrations, was linear between -40 and +30 mV. The steady-state conductance-voltage relationship was sigmoidal with membrane voltage, and, at pH 7.4, was one-half maximum at about +15 mV (V1/2). The time courses of activation and deactivation were proportional to 1-exp (-t/tau). A plot of time constant (tau) against voltage was bell-shaped, with a maximum near V1/2. These results suggested that the activation of IH is voltage dependent. Increases in the external H ion concentration shifted the conductance-voltage and time constant-voltage relations in parallel towards positive voltages. The magnitude of these shifts showed a lower saturation near pH 9. Low concentrations of external Cd (10-300 microM) reduced current amplitude by shifting the current-voltage relation in the positive direction. Cd also reduced the limiting slope conductance. These effects were partially reversible.
采用双微电极电压钳技术,对有尾两栖动物美西螈未成熟卵母细胞的膜电流进行了研究。由H离子携带的电流(IH)构成了从静息电压(约-60 mV)去极化激活的外向电流的主要部分。在这个发育阶段未观察到净内向电流。在pH 6.9至8.4之间,通过两步电压钳实验获得的尾电流测量的IH反转电位随外部pH每单位变化54 mV而移动。pH 7.4时的反转电位不受外部K或Cl浓度变化的影响。去除外部Na时观察到反转电位有小的变化。去除外部Ca(Mg或Sr替代)不影响IH的幅度。钙离子载体A23187使电流-电压关系向负电压方向移动。IH的激活似乎不依赖于Ca内流。在内部和外部H离子浓度大致相等时,从尾电流测量的IH瞬时电流-电压关系在-40至+30 mV之间呈线性。稳态电导-电压关系呈S形,与膜电压有关,在pH 7.4时,在约+15 mV(V1/2)处为最大值的一半。激活和失活的时间进程与1-exp(-t/tau)成正比。时间常数(tau)与电压的关系图呈钟形,在V1/2附近有最大值。这些结果表明IH的激活是电压依赖性的。外部H离子浓度的增加使电导-电压和时间常数-电压关系平行地向正电压方向移动。这些移动的幅度在pH 9附近显示出较低的饱和度。低浓度的外部Cd(10 - 300 microM)通过使电流-电压关系向正方向移动来降低电流幅度。Cd还降低了极限斜率电导。这些效应部分是可逆的。