Fisher R S, Van Driessche W
Department of Nephrology, Walter Reed Army Institute of Research, Washington, D.C. 20307-5100.
J Gen Physiol. 1991 Feb;97(2):219-43. doi: 10.1085/jgp.97.2.219.
We examined the development of K+ secretion after removing Cl- from the basolateral surface of isolated skins of Rana temporaria using noise analysis. K+ secretion was defined by the appearance of a Lorentzian component in the power density spectrum (PDS) when Ba2+ was present in the apical bath (0.5 mM). No Lorentzians were observed when tissues were bathed in control, NaCl Ringer solution. Replacement of basolateral Cl- by gluconate, nitrate, or SO4- (0-Clb) yielded Lorentzians with corner frequencies near 25 Hz, and plateau values (So) that were used to estimate the magnitude of K+ secretion through channels in the apical cell membranes of the principal cells. The response was reversible and reproducible. In contrast, removing apical Cl- did not alter the PDS. Reduction of basolateral Cl- to 11.5 mM induced Lorentzians, but with lower values of So. Inhibition of Na+ transport with amiloride or by omitting apical Na+ depressed K+ secretion but did not prevent its appearance in response to 0-Clb. Using microelectrodes, we observed depolarization of the intracellular voltage concomitant with increased resistance of the basolateral membrane after 0-Clb. Basolateral application of Ba2+ to depolarize cells also induced K+ secretion. Because apical conductance and channel density are unchanged after 0-Clb, we conclude that K+ secretion is "induced" simply by an increase of the electrical driving force for K+ exit across this membrane. Repolarization of the apical membrane after 0-Clb eliminated K+ secretion, while further depolarization increased the magnitude of the secretory current. The cell depolarization after 0-Clb is most likely caused directly by a decrease of the basolateral membrane K+ conductance. Ba2(+)-induced Lorentzians also were elicited by basolateral hypertonic solutions but with lower values of So, indicating that cell shrinkage per se could not entirely account for the response to 0-Clb and that the effects of 0-Clb may be partly related to a fall of intracellular Cl-.
我们使用噪声分析方法,研究了从林蛙离体皮肤的基底外侧表面去除氯离子后钾离子分泌的变化情况。当顶端浴液中存在钡离子(0.5 mM)时,通过功率密度谱(PDS)中洛伦兹分量的出现来定义钾离子分泌。当组织浸泡在对照的氯化钠林格溶液中时,未观察到洛伦兹分量。用葡萄糖酸盐、硝酸盐或硫酸根(0-Clb)替代基底外侧的氯离子,产生了拐角频率接近25 Hz的洛伦兹分量,以及用于估计通过主细胞顶端细胞膜通道的钾离子分泌量的平台值(So)。该反应是可逆且可重复的。相比之下,去除顶端的氯离子并未改变PDS。将基底外侧的氯离子浓度降低至11.5 mM会诱导洛伦兹分量的出现,但So值较低。用氨氯吡咪抑制钠离子转运或省略顶端的钠离子会抑制钾离子分泌,但并不能阻止其在响应0-Clb时出现。使用微电极,我们观察到在0-Clb处理后,细胞内电压去极化,同时基底外侧膜电阻增加。向基底外侧施加钡离子使细胞去极化也会诱导钾离子分泌。由于在0-Clb处理后顶端电导和通道密度未发生变化,我们得出结论,钾离子分泌仅仅是由钾离子跨膜流出的电驱动力增加所“诱导”的。0-Clb处理后顶端膜复极化消除了钾离子分泌,而进一步去极化则增加了分泌电流的幅度。0-Clb处理后细胞去极化很可能是直接由基底外侧膜钾离子电导降低引起的。基底外侧高渗溶液也能引发钡离子诱导的洛伦兹分量,但So值较低,这表明细胞本身收缩并不能完全解释对0-Clb的反应,且0-Clb的作用可能部分与细胞内氯离子浓度降低有关。