Wang W H, Wang Y, Silbernagl S, Oberleithner H
Department of Physiology, University of Würzburg, Federal Republic of Germany.
J Membr Biol. 1988 Mar;101(3):259-65. doi: 10.1007/BF01872840.
Experiments were performed in intact proximal tubules of the doubly perfused kidney and in fused proximal tubule cells of Rana esculenta to evaluate the dependence of intracellular pH (pHi) on cell membrane potential applying pH-sensitive and conventional microelectrodes. In proximal tubules an increase of the K+ concentration in the peritubular perfusate from 3 to 15 mmol/liter decreased the peritubular cell membrane potential from -55 +/- 2 to -38 +/- 1 mV paralleled by an increase of pHi from 7.54 +/- 0.02 to 7.66 +/- 0.02. The stilbene derivative DIDS hyperpolarized the cell membrane potential from -57 +/- 2 to -71 +/- 4 mV and led to a significant increase of the K+-induced cell membrane depolarization, but prevented the K+-induced intracellular alkalinization. Fused proximal tubule cells were impaled by three microelectrodes simultaneously and cell voltage was clamped stepwise while pHi changes were monitored. Cell membrane hyperpolarization acidified the cell cytoplasm in a linear relationship. This voltage-induced intracellular acidification was reduced to about one-third when HCO-3 ions were omitted from the extracellular medium. We conclude that in proximal tubule cells pHi depends on cell voltage due to the rheogenicity of the HCO-3 transport system.
实验在双灌注肾的完整近端小管以及食用蛙的融合近端小管细胞中进行,以使用pH敏感微电极和传统微电极评估细胞内pH(pHi)对细胞膜电位的依赖性。在近端小管中,管周灌流液中K +浓度从3 mmol/L增加到15 mmol/L,使管周细胞膜电位从-55±2 mV降低至-38±1 mV,同时pHi从7.54±0.02增加到7.66±0.02。芪衍生物DIDS使细胞膜电位从-57±2 mV超极化至-71±4 mV,并导致K +诱导的细胞膜去极化显著增加,但阻止了K +诱导的细胞内碱化。同时用三个微电极刺入融合的近端小管细胞,并逐步钳制细胞电压,同时监测pHi变化。细胞膜超极化使细胞质酸化呈线性关系。当细胞外培养基中省略HCO - 3离子时,这种电压诱导的细胞内酸化降低到约三分之一。我们得出结论,在近端小管细胞中,由于HCO - 3转运系统的生电性,pHi取决于细胞电压。