Frindt Gustavo, Palmer Lawrence G
Department of Physiology and Biophysics, Weill-Cornell Medical College, New York, New York.
Department of Physiology and Biophysics, Weill-Cornell Medical College, New York, New York
Am J Physiol Renal Physiol. 2015 Mar 15;308(6):F572-8. doi: 10.1152/ajprenal.00585.2014. Epub 2014 Dec 17.
The acute effects of aldosterone administration on epithelial Na channels (ENaC) in rat kidney were examined using electrophysiology and immunodetection. Animals received a single injection of aldosterone (20 μg/kg body wt), which reduced Na excretion over the next 3 h. Channel activity was assessed in principal cells of cortical collecting ducts as amiloride-sensitive whole cell clamp current (INa). INa averaged 100 pA/cell, 20-30% of that reported for the same preparation under conditions of chronic stimulation. INa was negligible in control animals that did not receive hormone. The acute physiological response correlated with changes in ENaC processing and trafficking. These effects included increases in the cleaved forms of α-ENaC and γ-ENaC, assessed by Western blot, and increases in the surface expression of β-ENaC and γ-ENaC measured after surface protein biotinylation. These changes were qualitatively and quantitatively similar to those of chronic stimulation. This suggests that altered trafficking to or from the apical membrane is an early response to the hormone and that later increases in channel activity require stimulation of channels residing at the surface.
采用电生理学和免疫检测方法,研究了醛固酮给药对大鼠肾脏上皮钠通道(ENaC)的急性影响。给动物单次注射醛固酮(20μg/kg体重),在接下来的3小时内钠排泄减少。在皮质集合管主细胞中,以氨氯地平敏感的全细胞钳电流(INa)评估通道活性。INa平均为100 pA/细胞,是相同制备物在慢性刺激条件下所报道值的20 - 30%。在未接受激素的对照动物中,INa可忽略不计。急性生理反应与ENaC加工和转运的变化相关。这些影响包括通过蛋白质免疫印迹法评估的α-ENaC和γ-ENaC裂解形式增加,以及表面蛋白生物素化后测量的β-ENaC和γ-ENaC表面表达增加。这些变化在性质和数量上与慢性刺激的变化相似。这表明向顶膜或从顶膜的转运改变是对该激素的早期反应,并且随后通道活性的增加需要刺激位于表面的通道。