Ishibashi K, Rector F C, Berry C A
Department of Physiology and Medicine, University of California, San Francisco 94143.
Am J Physiol. 1990 Jun;258(6 Pt 2):F1569-78. doi: 10.1152/ajprenal.1990.258.6.F1569.
To examine the basolateral Cl transport mechanisms of proximal convoluted tubules (PCT), intracellular Cl activity (AiCl) was measured with double-barreled Cl-selective microelectrodes. When rabbit PCT were perfused in vitro with high Cl, low HCO3, and bathed with ultrafiltrate-like solutions, AiCl was 29.9 +/- 0.4 mM and basolateral membrane voltage (Vbl) was -47.7 +/- 0.4 mV (n = 247). Possible basolateral Cl transport mechanisms that we examined were as follows: Cl conductance, KCl cotransport, and Na-dependent Cl-HCO3 exchange. Cl conductance was negligible, since the voltage clamp of Vbl to 30 mV above and below the spontaneous Vbl did not change AiCl even in the absence of luminal Cl. KCl cotransport was suggested by 1) increasing bath K, increased AiCl, and 2) decreasing bath K decreased AiCl. KCl cotransport was Na independent and 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS), barium, and furosemide insensitive. Na-dependent Cl-HCO3 exchange was suggested by 1) bath HCO3 reduction increased AiCl, which was greatly inhibited by bath Na removal or bath SITS, and 2) bath Na removal increased AiCl, which was completely blocked by bath SITS. We conclude that 1) Cl conductance is negligibly small at the basolateral membrane and 2) SITS-insensitive KCl cotransport and SITS-sensitive Na-dependent Cl-HCO3 exchange are present at the basolateral membrane.
为研究近端肾小管(PCT)基底外侧的氯离子转运机制,使用双管氯离子选择性微电极测量细胞内氯离子活性(AiCl)。当用高氯、低碳酸氢根的溶液对兔近端肾小管进行体外灌注,并浸泡于类似超滤液的溶液中时,AiCl为29.9±0.4 mM,基底外侧膜电压(Vbl)为-47.7±0.4 mV(n = 247)。我们研究的可能的基底外侧氯离子转运机制如下:氯离子电导、氯化钾共转运以及钠依赖的氯-碳酸氢根交换。氯离子电导可忽略不计,因为即使在管腔无氯离子的情况下,将Vbl钳制在自发Vbl上下30 mV时,AiCl也未发生变化。氯化钾共转运的依据为:1)增加浴液中的钾离子,AiCl增加;2)降低浴液中的钾离子,AiCl降低。氯化钾共转运不依赖于钠,且对4-乙酰氨基-4'-异硫氰基芪-2,2'-二磺酸(SITS)、钡和呋塞米不敏感。钠依赖的氯-碳酸氢根交换的依据为:1)降低浴液中的碳酸氢根可增加AiCl,而去除浴液中的钠或加入浴液SITS可显著抑制这一增加;2)去除浴液中的钠可增加AiCl,而加入浴液SITS可完全阻断这一增加。我们得出以下结论:1)基底外侧膜的氯离子电导极小;2)基底外侧膜存在对SITS不敏感的氯化钾共转运和对SITS敏感的钠依赖的氯-碳酸氢根交换。