Maintenance of extrecellular fluid volume homeostasis is essential for hemodynamic stability, and abnormalities of renal sodium handling have been linked to cardiovascular disease and hypertension. Ultimate regulation of sodium excretion occurs in the distal nephron via conductive transport through the amiloride sensitive epithelial Na+ channel (ENaC). ENaC expression and activity in the apical membrane of epithelial cells is the rate limiting step in Na+ reabsorption not only in the kidney collecting duct, but in airway epithelia and colon as well. Aldosterone is the major regulator of ENaC expression and activity in responsive epithelia. The long term objectives of this research are to define the mechanisms through which mineralocorticoid hormones regulate vectorial sodium transport in responsive epithelia and studies are designed to examine the mechanisms of aldosterone regulation of ENaC. Work in the current grant period has identified a specific methyltransferase which activates ENaC and knockdown of this protein blocks the early aldosterone response. Studies are now proposed to determine if this protein is required for early aldosterone action via genomic or non-genomic pathways and determine how aldosterone regulates its activity. We have demonstrated that ENaC trafficks to the apical membrane of epithelial cells in lipid rafts and developed a model of ENaC trafficking in cultured epithelial cells. Cells deprived of aldosterone develop a loss of Na+ reabsorptive activity along with a loss of the cellular recycling compartment for ENaC. The recycling compartment and Na+ reabsorptive phenotype are restored over time by aldosterone. Restoration of the ENaC recycling pathway is associated with adlosterone up-regulation of multiple proteins in the endosomal compartment. We will examine the mechanisms by which aldosterone regulates the apical expression of ENaC from endosomal and/or recycling pathways and determine what specific aldosterone-regulated proteins control the activity and expression of ENaC in endosomes and exocyst compartments to support apical translocation of ENaC in Na+ reabsorptive states.

Public Health Relevance

Aldosterone is the major hormone which regulates sodium reabsorption in the kidney in mammals. Abnormalities of sodium handling can lead to diseases such as hypertension and heart failure and have been implicated in the progression of cystic fibrosis lung disease. These studies are designed to determine the cellular mechanisms by which aldosterone regulates sodium reabsorption in kidney cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK047874-16
Application #
8514578
Study Section
Cellular and Molecular Biology of the Kidney Study Section (CMBK)
Program Officer
Ketchum, Christian J
Project Start
1995-05-01
Project End
2014-07-31
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
16
Fiscal Year
2013
Total Cost
$307,359
Indirect Cost
$101,648
Name
University of Pittsburgh
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Butterworth, Michael B; Edinger, Robert S; Silvis, Mark R et al. (2012) Rab11b regulates the trafficking and recycling of the epithelial sodium channel (ENaC). Am J Physiol Renal Physiol 302:F581-90
Edinger, Robert S; Lebowitz, Jonathan; Li, Hui et al. (2009) Functional regulation of the epithelial Na+ channel by IkappaB kinase-beta occurs via phosphorylation of the ubiquitin ligase Nedd4-2. J Biol Chem 284:150-7
Hallows, Kenneth R; Wang, Huamin; Edinger, Robert S et al. (2009) Regulation of epithelial Na+ transport by soluble adenylyl cyclase in kidney collecting duct cells. J Biol Chem 284:5774-83
Weixel, Kelly M; Edinger, Robert S; Kester, Lauren et al. (2007) Phosphatidylinositol 4-phosphate 5-kinase reduces cell surface expression of the epithelial sodium channel (ENaC) in cultured collecting duct cells. J Biol Chem 282:36534-42
Butterworth, Michael B; Edinger, Robert S; Ovaa, Huib et al. (2007) The deubiquitinating enzyme UCH-L3 regulates the apical membrane recycling of the epithelial sodium channel. J Biol Chem 282:37885-93
Hill, Warren G; Butterworth, Michael B; Wang, Huamin et al. (2007) The epithelial sodium channel (ENaC) traffics to apical membrane in lipid rafts in mouse cortical collecting duct cells. J Biol Chem 282:37402-11
Edinger, Robert S; Yospin, Jeremy; Perry, Clint et al. (2006) Regulation of epithelial Na+ channels (ENaC) by methylation: a novel methyltransferase stimulates ENaC activity. J Biol Chem 281:9110-7
Carattino, Marcelo D; Edinger, Robert S; Grieser, Heather J et al. (2005) Epithelial sodium channel inhibition by AMP-activated protein kinase in oocytes and polarized renal epithelial cells. J Biol Chem 280:17608-16
Lebowitz, Jonathan; Edinger, Robert S; An, Bing et al. (2004) Ikappab kinase-beta (ikkbeta) modulation of epithelial sodium channel activity. J Biol Chem 279:41985-90
Wang, Edward C Y; Lee, Jey-Myung; Johnson, John P et al. (2003) Hydrostatic pressure-regulated ion transport in bladder uroepithelium. Am J Physiol Renal Physiol 285:F651-63

Showing the most recent 10 out of 21 publications