Expression of the chloride channel CLC-K in human airway epithelial cells

Jennifer L. Mummery, Jennifer Killey, Paul Linsdell

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Airway submucosal gland function is severely disrupted in cystic fibrosis (CF), as a result of genetic mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), an apical membrane Cl- channel. To identify other Cl- channel types that could potentially substitute for lost CFTR function in these cells, we investigated the functional and molecular expression of Cl- channels in Calu-3 cells, a human cell line model of the submucosal gland serous cell. Whole cell patch clamp recording from these cells identified outwardly rectified, pH- and calcium-sensitive Cl- currents that resemble those previously ascribed to ClC-K type chloride channels. Using reverse transcription - polymerase chain reaction, we identified expression of mRNA for ClC-2, ClC-3, ClC-4, ClC-5, ClC-6, ClC-7, ClC-Ka, and ClC-Kb, as well as the common ClC-K channel β subunit barttin. Western blotting confirmed that Calu-3 cells express both ClC-K and barttin protein. Thus, Calu-3 cells express multiple members of the ClC family of Cl- channels that, if also expressed in native submucosal gland serous cells within the CF lung, could perhaps act to partially substitute lost CFTR function. Furthermore, this work represents the first evidence for functional ClC-K chloride channel expression within the lung.

LanguageEnglish
Pages1123-1128
Number of pages6
JournalCanadian Journal of Physiology and Pharmacology
Volume83
Issue number12
DOIs
Publication statusPublished - Dec 2005

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Chloride Channels
Epithelial Cells
Cystic Fibrosis Transmembrane Conductance Regulator
Cystic Fibrosis
Lung
Ion Channels
Reverse Transcription
Western Blotting
Calcium
Cell Line
Polymerase Chain Reaction
Messenger RNA
Mutation

Cite this

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abstract = "Airway submucosal gland function is severely disrupted in cystic fibrosis (CF), as a result of genetic mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), an apical membrane Cl- channel. To identify other Cl- channel types that could potentially substitute for lost CFTR function in these cells, we investigated the functional and molecular expression of Cl- channels in Calu-3 cells, a human cell line model of the submucosal gland serous cell. Whole cell patch clamp recording from these cells identified outwardly rectified, pH- and calcium-sensitive Cl- currents that resemble those previously ascribed to ClC-K type chloride channels. Using reverse transcription - polymerase chain reaction, we identified expression of mRNA for ClC-2, ClC-3, ClC-4, ClC-5, ClC-6, ClC-7, ClC-Ka, and ClC-Kb, as well as the common ClC-K channel β subunit barttin. Western blotting confirmed that Calu-3 cells express both ClC-K and barttin protein. Thus, Calu-3 cells express multiple members of the ClC family of Cl- channels that, if also expressed in native submucosal gland serous cells within the CF lung, could perhaps act to partially substitute lost CFTR function. Furthermore, this work represents the first evidence for functional ClC-K chloride channel expression within the lung.",
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Expression of the chloride channel CLC-K in human airway epithelial cells. / Mummery, Jennifer L.; Killey, Jennifer; Linsdell, Paul.

In: Canadian Journal of Physiology and Pharmacology, Vol. 83, No. 12, 12.2005, p. 1123-1128.

Research output: Contribution to journalArticle

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AB - Airway submucosal gland function is severely disrupted in cystic fibrosis (CF), as a result of genetic mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), an apical membrane Cl- channel. To identify other Cl- channel types that could potentially substitute for lost CFTR function in these cells, we investigated the functional and molecular expression of Cl- channels in Calu-3 cells, a human cell line model of the submucosal gland serous cell. Whole cell patch clamp recording from these cells identified outwardly rectified, pH- and calcium-sensitive Cl- currents that resemble those previously ascribed to ClC-K type chloride channels. Using reverse transcription - polymerase chain reaction, we identified expression of mRNA for ClC-2, ClC-3, ClC-4, ClC-5, ClC-6, ClC-7, ClC-Ka, and ClC-Kb, as well as the common ClC-K channel β subunit barttin. Western blotting confirmed that Calu-3 cells express both ClC-K and barttin protein. Thus, Calu-3 cells express multiple members of the ClC family of Cl- channels that, if also expressed in native submucosal gland serous cells within the CF lung, could perhaps act to partially substitute lost CFTR function. Furthermore, this work represents the first evidence for functional ClC-K chloride channel expression within the lung.

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