1. Effects of mineralocorticoid and K+ concentration on K+ secretion and ROMK channel expression in a mouse cortical collecting duct cell line.
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Fodstad, Heidi, Gonzalez-Rodriguez, Elena, Bron, Sylvian, Gaeggeler, Hanspeter, Guisan, Barbara, Rossier, Bernard C., and Horisberger, Jean-Daniel
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CELLULAR mechanics ,MESSENGER RNA ,CHEMICAL reactions ,MINERALOCORTICOIDS ,AMILORIDE ,ALDOSTERONE - Abstract
The cortical collecting duct (CCD) plays a key role in regulated K
+ secretion, which is mediated mainly through renal outer medullary K+ (ROMK) channels located in the apical membrane. However, the mechanisms of the regulation of urinary K+ excretion with regard to K+ balance are not well known. We took advantage of a recently established mouse CCD cell line (mCCDc11 ) to investigate the regulation of K+ secretion by mineralocorticoid and K+ concentration. We show that this cell line expresses ROMK mRNA and a barium-sensitive K+ conductance in its apical membrane. As this conductance is sensitive to tertiapin-Q, with an apparent affinity of 6 nM, and to intracelIular acidification, it is probably mediated by ROMK. Overnight exposure to 100 nM aldosterone did not significantly change the K+ conductance, while it increased the amiloride-sensitive Na+ transport. Overnight exposure to a high K+ (7 mM) concentration produced a small but significant increase in the apical membrane barium-sensitive K+ conductance. The mRNA levels of all ROMK isoforms measured by qRT-PCR were not changed by altering the basolateral K+ concentration but were decreased by 15-45% upon treatment with aldosterone (0.3 or 300 nM for 1 and 3 h). The paradoxical response of ROMK expression to aldosterone could possibly work as a preventative mechanism to avoid excessive K+ loss which would otherwise result from the increased electrogenic Na+ transport and associated depolarization of the apical membrane in the CCD. In conclusion, mCCDc11 cells demonstrate a significant K+ secretion, probably mediated by ROMK, which is not stimulated by aldosterone but increased by overnight exposure to a high K+ concentration. [ABSTRACT FROM AUTHOR]- Published
- 2009
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