Biagi B, Kubota T, Sohtell M, Giebisch G
Am J Physiol. 1981 Mar;240(3):F200-10. doi: 10.1152/ajprenal.1981.240.3.F200.
Conventional microelectrodes were used to measure the basolateral membrane potential (VBL) in isolated perfused superficial proximal convoluted (sPCT) and superficial proximal straight (sPST) tubules of the rabbit kidney. Stable recordings for periods up to 2 h can be obtained. The mean +/- SE (n = number of cells) values of VBL were sPCT = -51.0 +/- 1.63 (24) and sPST = -47.0 +/- 0.97 (94) mV. Inhibitors of active transport, ouabain (10(-5) M) and low bath potassium (0.1 mM), caused a significant depolarization of VBL in sPST. In contrast, short-duration bath cooling (10 degrees C) had no significant effect. Removal of luminal glucose caused a larger hyperpolarization in sPCT (-13.9 +/- 1.77 (9) mV) than in sPST (-3.8 +/- 1.02 (5) mV). Removal of luminal glucose and alanine resulted in an even larger hyperpolarization of VBL in sPCT (-19.0 +/- 0.44 (6) mV). Perfusion of the lumen with a solution resembling late proximal tubular fluid in sPST resulted in hyperpolarization of VBL (-4.3 +/- 0.85 (4) mV). Reducing bath pH to 6.7 depolarized VBL (39.9 +/- 1.77 (13) mV). This effect can be associated with a decrease in the relative potassium permeability of the basolateral membrane. These results demonstrate the feasibility of using intracellular electrical measurements to determine both luminal and basolateral membrane characteristics in isolated proximal tubular segments.
采用传统微电极测量兔肾分离灌注的浅表近端曲管(sPCT)和浅表近端直小管(sPST)的基底外侧膜电位(VBL)。可获得长达2小时的稳定记录。VBL的平均±标准误(n =细胞数量)值为:sPCT = -51.0±1.63(24)mV,sPST = -47.0±0.97(94)mV。主动转运抑制剂哇巴因(10⁻⁵ M)和低浴钾(0.1 mM)可导致sPST中VBL显著去极化。相比之下,短时间浴温冷却(10℃)无显著影响。去除管腔葡萄糖导致sPCT中(-13.9±1.77(9)mV)的超极化幅度大于sPST中(-3.8±1.02(5)mV)。去除管腔葡萄糖和丙氨酸导致sPCT中VBL出现更大幅度的超极化(-19.0±0.44(6)mV)。用类似于sPST中晚期近端小管液的溶液灌注管腔导致VBL超极化(-4.3±0.85(4)mV)。将浴液pH降至6.7使VBL去极化(39.9±1.77(13)mV)。这种效应可能与基底外侧膜相对钾通透性降低有关。这些结果证明了使用细胞内电测量来确定分离的近端小管节段管腔和基底外侧膜特性的可行性。