O'Neil R G, Sansom S C
J Membr Biol. 1984;82(3):281-95. doi: 10.1007/BF01871637.
Microelectrode techniques were applied to the rabbit isolated perfused cortical collecting duct to provide an initial quantitation and characterization of the cell membrane and tight junction conductances. Initial studies demonstrated that the fractional resistance (ratio of the resistance of the apical cell membrane to the sum of the resistances of the apical and basolateral membranes) was usually independent of the point along the tubule of microelectrode impalement--implicating little cell-to-cell coupling--supporting the application of quantitative techniques to the cortical collecting duct. It was demonstrated that in the presence of amiloride, either reduction in the luminal pH or the addition of barium to the perfusate selectively reduced the apical membrane potassium conductance. From the changes in Gte and fractional resistance upon reducing the luminal pH or addition of barium to the perfusate, the transepithelial, apical membrane, basolateral membrane and tight junction conductances were estimated to be 9.3, 6.7, 8.1 and 6.0 mS cm-2, respectively. Ninety to ninety-five percent of the apical membrane conductance reflected the barium-sensitive potassium conductance in the presence of amiloride with an estimated potassium permeability of 1.1 X 10(-4) cm sec-1. Reduction in the perfusate pH to 4.0 caused a 70% decrease in the apical membrane potassium conductance, implying a blocking site with an acidic group having a pKa near 4.4. It is concluded that both the transcellular and paracellular pathways of the cortical collecting tubule have high ionic conductances, and that the apical membrane conductance primarily reflects a high potassium conductance. Furthermore, both reduction in the perfusate pH and addition of barium to the perfusate selectively block the apical potassium channels, although the site of inhibition likely differs since the two ions display markedly different voltage-dependent blocks of the channel.
将微电极技术应用于兔离体灌注皮质集合管,以初步定量和表征细胞膜及紧密连接的电导。初步研究表明,分数电阻(顶端细胞膜电阻与顶端和基底外侧膜电阻之和的比值)通常与微电极刺入肾小管的位置无关——这意味着细胞间耦合较少——支持将定量技术应用于皮质集合管。结果表明,在存在氨氯吡咪的情况下,管腔pH值降低或向灌注液中添加钡会选择性地降低顶端膜钾电导。根据降低管腔pH值或向灌注液中添加钡时Gte和分数电阻的变化,估计跨上皮、顶端膜、基底外侧膜和紧密连接的电导分别为9.3、6.7、8.1和6.0 mS cm-2。在存在氨氯吡咪的情况下,90%至95%的顶端膜电导反映了对钡敏感的钾电导,估计钾通透性为1.1×10(-4) cm sec-1。将灌注液pH值降至4.0会导致顶端膜钾电导降低70%,这意味着存在一个pKa接近4.4的带酸性基团的阻断位点。结论是,皮质集合小管的跨细胞和细胞旁途径都具有高离子电导,并且顶端膜电导主要反映高钾电导。此外,灌注液pH值降低和向灌注液中添加钡都会选择性地阻断顶端钾通道,尽管抑制位点可能不同,因为这两种离子对通道的电压依赖性阻断明显不同。