The kidney maintains the balance of potassium (K) in the body by excreting in the urine about the same amount of K that is ingested in the diet. Under most conditions it does this by secreting K into the urine in the distal nephron and collecting duct. The mechanism involves, at least in part, the movement of K from the cells into the urine through ion channels in the apical membrane. The connecting tubule (CNT), a segment joining the distal convoluted tubule and the cortical collecting duct (CCD) is an important site for K secretion. We have identified two K channels in the apical membrane of the rat CNT: a low conductance (SK, ROMK) channel and a high conductance (BK, maxi-K) channel. Here we propose to examine the regulation of these channels in the CNT. In our initial findings the abundance of SK channels was not increased in the CNT as it was in the CCD. We will test whether the set-point for regulation is shifted in the CNT. Alternatively, we will examine whether the difference is related to the ROMK isoform which is expressed. We will also examine the regulation of the SK channels by peptide hormones including ADH and glucagon. The BK channels were observed primarily in the intercalated cells of the CNT. We will ask whether the low open probability of these channels under resting conditions can be increased sufficiently for them to contribute to K secretion. Stimuli will include hormone stimulation and increased shear stress on the membrane. We will also ask whether K uptake by the intercalated cells through the Na/K-ATPase is adequate to support a significant K secretion. Finally, we will quantify K secretion in the distal nephron under various conditions and ask whether it can be accounted for by the measured electrophysiological properties of the cells. A second set of aims will focus on the process of permeation through SK/ROMK channels. We will test the hypothesis that this K movement involves transport through the """"""""cytoplasmic pore"""""""", a part of the channel protein which protrudes into the cytoplasm and is unique to the inward-rectifier K channel family. We will further examine the mechanism of saturation of channel conductance using a site-directed mutagenesis approach. These studies should elucidate the processes of how the kidney maintains plasma K under tight control. Loss of this control, for example in genetic diseases such as Bartter's or Gitelman's syndromes, has serious consequences including altered cell excitability, cardiac arrhythmias and muscle weakness. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK027847-24
Application #
7047650
Study Section
Special Emphasis Panel (ZRG1-RUS-D (02))
Program Officer
Ketchum, Christian J
Project Start
1981-03-01
Project End
2010-01-31
Budget Start
2006-02-01
Budget End
2007-01-31
Support Year
24
Fiscal Year
2006
Total Cost
$377,241
Indirect Cost
Name
Weill Medical College of Cornell University
Department
Physiology
Type
Schools of Medicine
DUNS #
060217502
City
New York
State
NY
Country
United States
Zip Code
10065
Yang, Lei; Frindt, Gustavo; Lang, Florian et al. (2017) SGK1-dependent ENaC processing and trafficking in mice with high dietary K intake and elevated aldosterone. Am J Physiol Renal Physiol 312:F65-F76
Frindt, Gustavo; Gravotta, Diego; Palmer, Lawrence G (2016) Regulation of ENaC trafficking in rat kidney. J Gen Physiol 147:217-27
Frindt, Gustavo; Palmer, Lawrence G (2015) Acute effects of aldosterone on the epithelial Na channel in rat kidney. Am J Physiol Renal Physiol 308:F572-8
Palmer, Lawrence G; Schnermann, J├╝rgen (2015) Integrated control of Na transport along the nephron. Clin J Am Soc Nephrol 10:676-87
Patel, Ankit B; Yang, Lei; Deng, Su et al. (2014) Feedback inhibition of ENaC: acute and chronic mechanisms. Channels (Austin) 8:444-51
Yang, Lei; Palmer, Lawrence G (2014) Ion conduction and selectivity in acid-sensing ion channel 1. J Gen Physiol 144:245-55
Frindt, Gustavo; Li, Hui; Sackin, Henry et al. (2013) Inhibition of ROMK channels by low extracellular K+ and oxidative stress. Am J Physiol Renal Physiol 305:F208-15
Yang, Lei; Edvinsson, Johan; Sackin, Henry et al. (2012) Ion selectivity and current saturation in inward-rectifier K+ channels. J Gen Physiol 139:145-57
Yang, Lei; Edvinsson, Johan; Palmer, Lawrence G (2012) Interactions of external K+ and internal blockers in a weak inward-rectifier K+ channel. J Gen Physiol 140:529-40
Sackin, Henry; Nanazashvili, Mikheil; Li, Hui et al. (2012) Residues at the outer mouth of Kir1.1 determine K-dependent gating. Biophys J 102:2742-50

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