Spring K R, Paganelli C V
J Gen Physiol. 1972 Aug;60(2):181-201. doi: 10.1085/jgp.60.2.181.
Na transport and electrical properties of Necturus renal proximal tubules were analyzed, in vivo, by a voltage clamp method which utilizes an axial electrode in the tubule lumen for passage of current and simultaneous determination of net fluid (or Na) flux by the split droplet method. When the average spontaneous transepithelial potential difference of -8 mv (lumen negative) was reduced to zero by current passage, net Na flux doubled from a mean of 107 to 227 pmoles/cm(2) per sec. The relationship between flux and potential over the range -25 to +10 mv was nonlinear, with flux equilibrium at -15 mv and droplet expansion at more negative values. Calculated Na permeability at flux equilibrium was 7.0 x 10(-6) cm/sec. Voltage transients, similar to those caused by intraepithelial unstirred layers, were observed at the end of clamping periods. Tubular electrical resistance measured by brief square or triangle wave pulses (<100 msec) averaged 43 ohm cm(2). The epithelial current-voltage relationship was linear over the range -100 to +100 mv, but displayed marked hysteresis during low frequency (<0.04 Hz) triangle wave clamps. The low transepithelial resistance and large opposing unidirectional ion fluxes suggest that passive ionic movements occur across extracellular shunt pathways, while the voltage transients and current-voltage hysteresis are consistent with the development of a local osmotic gradient within epithelium.
采用电压钳法在体内分析了美西螈肾近端小管的钠转运和电特性,该方法利用小管腔内的轴向电极来通过电流,并同时用分滴法测定净液体(或钠)通量。当通过电流使平均-8 mV(管腔为负)的自发跨上皮电位差降至零时,净钠通量从平均每秒107皮摩尔/平方厘米增加一倍至227皮摩尔/平方厘米。在-25至+10 mV范围内,通量与电位之间的关系是非线性的,通量在-15 mV时达到平衡,在更负值时液滴扩张。通量平衡时计算出的钠渗透率为7.0×10⁻⁶厘米/秒。在钳制期结束时观察到电压瞬变,类似于由上皮内未搅动层引起的瞬变。通过短暂的方波或三角波脉冲(<100毫秒)测量的小管电阻平均为43欧姆·平方厘米。上皮电流-电压关系在-100至+100 mV范围内是线性的,但在低频(<0.04 Hz)三角波钳制期间表现出明显的滞后现象。低跨上皮电阻和大的反向单向离子通量表明被动离子运动通过细胞外旁路途径发生,但电压瞬变和电流-电压滞后与上皮内局部渗透梯度形成一致。