Palmer L G, Antonian L, Frindt G
Department of Physiology and Biophysics, Cornell University Medical College, New York 10021.
J Gen Physiol. 1993 Jul;102(1):43-57. doi: 10.1085/jgp.102.1.43.
Activities of Na channels and Na pumps were studied in the rat cortical collecting tubule (CCT) during manipulation of the animals' mineralocorticoid status in vivo using a low-Na diet, diuretics, or administration of exogenous aldosterone. Tubules were isolated and split open to expose the luminal membrane surface. Using the whole-cell patch-clamp technique, activities of the apical Na channels and the basolateral Na pumps were measured in principal cells as the currents inhibited by amiloride (10 microM) and ouabain (1 mM), respectively. Na channel current (INa) was not measurable in CCTs from control animals on a normal diet. INa was approximately 200 pA/cell in CCTs from animals on a low-Na diet or infused with aldosterone using osmotic minipumps. Currents attributable to the Na pump (Ipump) were similar in control animals and animals on a low-Na diet. Maximal currents were approximately 35 pA/cell in both groups, and decreased with hyperpolarization of the cell membrane. In contrast, administration of exogenous aldosterone increased Ipump fourfold. Coinfusion of aldosterone and amiloride in vivo through the minipumps did not affect the induction of INa but reduced the induction of Ipump by 80%. We conclude that the induction of channel activity in this tissue is a direct action of aldosterone, whereas the induction of pump activity may be a consequence of the increased Na traffic through the epithelial cells.
在体内通过低钠饮食、利尿剂或给予外源性醛固酮来改变动物的盐皮质激素状态时,对大鼠皮质集合管(CCT)中的钠通道和钠泵活性进行了研究。分离出肾小管并将其切开以暴露管腔膜表面。使用全细胞膜片钳技术,分别测量主细胞顶端钠通道和基底外侧钠泵的活性,即分别测量被氨氯吡脒(10微摩尔)和哇巴因(1毫摩尔)抑制的电流。正常饮食的对照动物的CCT中无法测量钠通道电流(INa)。低钠饮食动物或使用渗透微型泵注入醛固酮的动物的CCT中,INa约为200皮安/细胞。对照动物和低钠饮食动物中归因于钠泵的电流(Ipump)相似。两组的最大电流均约为35皮安/细胞,并随着细胞膜的超极化而降低。相比之下,给予外源性醛固酮使Ipump增加了四倍。通过微型泵在体内同时注入醛固酮和氨氯吡脒并不影响INa的诱导,但使Ipump的诱导降低了80%。我们得出结论,该组织中通道活性的诱导是醛固酮的直接作用,而泵活性的诱导可能是钠通过上皮细胞流量增加的结果。