Differential regulation of ROMK (Kir1.1) in distal nephron segments by dietary potassium

James B. Wade, Liang Fang, Richard A. Coleman, Jie Liu, P. Richard Grimm, Tong Wang, Paul A. Welling

Research output: Contribution to journalArticle

Abstract

ROMK channels are well-known to play a central role in renal K secretion, but the absence of highly specific and avid-ROMK antibodies has presented significant roadblocks toward mapping the extent of expression along the entire distal nephron and determining whether surface density of these channels is regulated in response to physiological stimuli. Here, we prepared new ROMK antibodies verified to be highly specific, using ROMK knockout mice as a control. Characterization with segmental markers revealed a more extensive pattern of ROMK expression along the entire distal nephron than previously thought, localizing to distal convoluted tubule regions, DCT1 and DCT2; the connecting tubule (CNT); and cortical collecting duct (CD). ROMK was diffusely distributed in intracellular compartments and at the apical membrane of each tubular region. Apical labeling was significantly increased by high-K diet in DCT2, CNT1, CNT2, and CD (P < 0.05) but not in DCT1. Consistent with the large increase in apical ROMK, dramatically increased mature glycosylation was observed following dietary potassium augmentation. We conclude 1) our new antibody provides a unique tool to characterize ROMK channel localization and expression and 2) high-K diet causes a large increase in apical expression of ROMK in DCT2, CNT, and CD but not in DCT1, indicating that different regulatory mechanisms are involved in K diet-regulated ROMK channel functions in the distal nephron.

Original languageEnglish (US)
Pages (from-to)F1385-F1393
JournalAmerican Journal of Physiology - Renal Physiology
Volume300
Issue number6
DOIs
StatePublished - Jun 1 2011
Externally publishedYes

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Dietary Potassium
Nephrons
Diet
Antibodies
Glycosylation
Knockout Mice
Kidney
Membranes

Keywords

  • ROMK antibody
  • ROMK localization

ASJC Scopus subject areas

  • Physiology
  • Urology

Cite this

Differential regulation of ROMK (Kir1.1) in distal nephron segments by dietary potassium. / Wade, James B.; Fang, Liang; Coleman, Richard A.; Liu, Jie; Richard Grimm, P.; Wang, Tong; Welling, Paul A.

In: American Journal of Physiology - Renal Physiology, Vol. 300, No. 6, 01.06.2011, p. F1385-F1393.

Research output: Contribution to journalArticle

Wade, James B. ; Fang, Liang ; Coleman, Richard A. ; Liu, Jie ; Richard Grimm, P. ; Wang, Tong ; Welling, Paul A. / Differential regulation of ROMK (Kir1.1) in distal nephron segments by dietary potassium. In: American Journal of Physiology - Renal Physiology. 2011 ; Vol. 300, No. 6. pp. F1385-F1393.
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