Novel role for pendrin in orchestrating bicarbonate secretion in cystic fibrosis transmembrane conductance regulator (CFTR)-expressing airway serous cells

James P. Garnett, Emma Hickman, Rachel Burrows, P. Hegyi, L. Tiszlavicz, Alan W. Cuthbert, Peying Fong, Michael A. Gray

Research output: Contribution to journalArticle

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Abstract

In most HCO 3 --secreting epithelial tissues, SLC26 Cl -/HCO 3 - transporters work in concert with the cystic fibrosis transmembrane conductance regulator (CFTR) to regulate the magnitude and composition of the secreted fluid, a process that is vital for normal tissue function. By contrast, CFTR is regarded as the only exit pathway for HCO 3 - in the airways. Here we show that Cl -/HCO 3 - anion exchange makes a major contribution to transcellular HCO 3 - transport in airway serous cells. Real-time measurement of intracellular pH from polarized cultures of human Calu-3 cells demonstrated cAMP/PKA-activated Cl --dependent HCO 3 - transport across the luminal membrane via CFTR-dependent coupled Cl -/HCO 3 - anion exchange. The pharmacological and functional profile of the luminal anion exchanger was consistent with SLC26A4 (pendrin), which was shown to be expressed by quantitative RT-PCR, Western blot, and immunofluorescence. Pendrin-mediated anion exchange activity was confirmed by shRNA pendrin knockdown (KD), which markedly reduced cAMP-activated Cl -/HCO 3 - exchange. To establish the relative roles of CFTR and pendrin in net HCO 3 - secretion, transepithelial liquid secretion rate and liquid pH were measured in wild type, pendrin KD, and CFTR KD cells. cAMP/PKA increased the rate and pH of the secreted fluid. Inhibiting CFTR reduced the rate of liquid secretion but not the pH, whereas decreasing pendrin activity lowered pH with little effect on volume. These results establish that CFTR predominately controls the rate of liquid secretion, whereas pendrin regulates the composition of the secreted fluid and identifies a critical role for this anion exchanger in transcellular HCO 3 - secretion in airway serous cells.

Original languageEnglish
Pages (from-to)41069-41082
Number of pages14
JournalJournal of Biological Chemistry
Volume286
Issue number47
DOIs
Publication statusPublished - Nov 25 2011

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Cystic Fibrosis Transmembrane Conductance Regulator
Bicarbonates
Anions
Liquids
Fluids
Tissue
Time measurement
Chemical analysis
Small Interfering RNA
Fluorescent Antibody Technique
Epithelium
Western Blotting
Pharmacology
Membranes
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Novel role for pendrin in orchestrating bicarbonate secretion in cystic fibrosis transmembrane conductance regulator (CFTR)-expressing airway serous cells. / Garnett, James P.; Hickman, Emma; Burrows, Rachel; Hegyi, P.; Tiszlavicz, L.; Cuthbert, Alan W.; Fong, Peying; Gray, Michael A.

In: Journal of Biological Chemistry, Vol. 286, No. 47, 25.11.2011, p. 41069-41082.

Research output: Contribution to journalArticle

Garnett, James P. ; Hickman, Emma ; Burrows, Rachel ; Hegyi, P. ; Tiszlavicz, L. ; Cuthbert, Alan W. ; Fong, Peying ; Gray, Michael A. / Novel role for pendrin in orchestrating bicarbonate secretion in cystic fibrosis transmembrane conductance regulator (CFTR)-expressing airway serous cells. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 47. pp. 41069-41082.
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AB - In most HCO 3 --secreting epithelial tissues, SLC26 Cl -/HCO 3 - transporters work in concert with the cystic fibrosis transmembrane conductance regulator (CFTR) to regulate the magnitude and composition of the secreted fluid, a process that is vital for normal tissue function. By contrast, CFTR is regarded as the only exit pathway for HCO 3 - in the airways. Here we show that Cl -/HCO 3 - anion exchange makes a major contribution to transcellular HCO 3 - transport in airway serous cells. Real-time measurement of intracellular pH from polarized cultures of human Calu-3 cells demonstrated cAMP/PKA-activated Cl --dependent HCO 3 - transport across the luminal membrane via CFTR-dependent coupled Cl -/HCO 3 - anion exchange. The pharmacological and functional profile of the luminal anion exchanger was consistent with SLC26A4 (pendrin), which was shown to be expressed by quantitative RT-PCR, Western blot, and immunofluorescence. Pendrin-mediated anion exchange activity was confirmed by shRNA pendrin knockdown (KD), which markedly reduced cAMP-activated Cl -/HCO 3 - exchange. To establish the relative roles of CFTR and pendrin in net HCO 3 - secretion, transepithelial liquid secretion rate and liquid pH were measured in wild type, pendrin KD, and CFTR KD cells. cAMP/PKA increased the rate and pH of the secreted fluid. Inhibiting CFTR reduced the rate of liquid secretion but not the pH, whereas decreasing pendrin activity lowered pH with little effect on volume. These results establish that CFTR predominately controls the rate of liquid secretion, whereas pendrin regulates the composition of the secreted fluid and identifies a critical role for this anion exchanger in transcellular HCO 3 - secretion in airway serous cells.

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