ROMK (Kir1.1) forms apical K channels in the distal nephron and collecting duct that provide the K secretory pathways for 1) K recycling necessary for NaCI absorption by the thick ascending limb and 2) K secretion by connecting duct and principal cells. Therefore, defining the function and regulation of the ROMK channels is important for understanding renal K handling and body K homeostasis. Loss-of-function mutations in ROMK cause Bartter's syndrome, a hypotensive renal salt wasting disease, that is due to loss of salt reabsorbing capacity by the thick ascending limb. We have developed the only ROMK Bartter's mouse with sufficiently high survival (>30%) to permit physiological study - and is the only mouse model of Bartter's with such high survival. We propose to continue to study the pathophysiology of mouse ROMK Bartter's to enhance our understanding of the role of ROMK in renal K handling and to suggest potential modalities for therapy of human Bartter's syndrome. We will also develop two new ROMK transgenic mice that will provide models for assessing both the specific roles of ROMK isoforms and the regulated trafficking of ROMK isoforms in native kidney cells. One of these isoforms, ROMK1, is only expressed in late distal tubule and collecting duct and has been suggested to play an important role in the regulation of renal K secretion by dietary K intake. We propose to generate a mouse deficient only in ROMK1 by selective deletion of the ROMK1-specific exon using a Cre-LoxP strategy. We hypothesize that this mouse will not have a Bartter's phenotype, but instead, have disordered K handling with increases or decreases in dietary K intake. We will also generate a ROMK1 BAC-transgenic mouse tagged with FLAG and EGFP to define ROMK trafficking in kidney epithelial cells and to determine associated proteins in the ROMK regulatory complex. These mice will also provide a resource for others interested in ROMK function in non- renal cells (Gl tract, brain, etc).

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK054999-07
Application #
7230528
Study Section
Cellular and Molecular Biology of the Kidney Study Section (CMBK)
Program Officer
Ketchum, Christian J
Project Start
2001-03-01
Project End
2009-05-31
Budget Start
2007-06-01
Budget End
2008-05-31
Support Year
7
Fiscal Year
2007
Total Cost
$329,459
Indirect Cost
Name
Yale University
Department
Physiology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Dong, Ke; Yan, Qingshang; Lu, Ming et al. (2016) Romk1 Knockout Mice Do Not Produce Bartter Phenotype but Exhibit Impaired K Excretion. J Biol Chem 291:5259-69
Vucic, Esad; Alfadda, Tariq; MacGregor, Gordon G et al. (2015) Kir1.1 (ROMK) and Kv7.1 (KCNQ1/KvLQT1) are essential for normal gastric acid secretion: importance of functional Kir1.1. Pflugers Arch 467:1457-1468
Wade, James B; Fang, Liang; Coleman, Richard A et al. (2011) Differential regulation of ROMK (Kir1.1) in distal nephron segments by dietary potassium. Am J Physiol Renal Physiol 300:F1385-93
Geibel, John P; Hebert, Steven C (2009) The functions and roles of the extracellular Ca2+-sensing receptor along the gastrointestinal tract. Annu Rev Physiol 71:205-17
Yan, Qingshang; Yang, Xinbo; Cantone, Alessandra et al. (2008) Female ROMK null mice manifest more severe Bartter II phenotype on renal function and higher PGE2 production. Am J Physiol Regul Integr Comp Physiol 295:R997-R1004
Cantone, Alessandra; Yang, Xinbo; Yan, Qingshang et al. (2008) Mouse model of type II Bartter's syndrome. I. Upregulation of thiazide-sensitive Na-Cl cotransport activity. Am J Physiol Renal Physiol 294:F1366-72
Wagner, Carsten A; Loffing-Cueni, Dominique; Yan, Qingshang et al. (2008) Mouse model of type II Bartter's syndrome. II. Altered expression of renal sodium- and water-transporting proteins. Am J Physiol Renal Physiol 294:F1373-80
Lu, Ming; Leng, Qiang; Egan, Marie E et al. (2006) CFTR is required for PKA-regulated ATP sensitivity of Kir1.1 potassium channels in mouse kidney. J Clin Invest 116:797-807
Leng, Qiang; MacGregor, Gordon G; Dong, Ke et al. (2006) Subunit-subunit interactions are critical for proton sensitivity of ROMK: evidence in support of an intermolecular gating mechanism. Proc Natl Acad Sci U S A 103:1982-7
Bailey, M A; Cantone, A; Yan, Q et al. (2006) Maxi-K channels contribute to urinary potassium excretion in the ROMK-deficient mouse model of Type II Bartter's syndrome and in adaptation to a high-K diet. Kidney Int 70:51-9

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