Abstract
Salt and water absorption and secretion across the airway epithelium are important for maintaining the thin film of liquid lining the surface of the airway epithelium. Movement of Cl across the apical membrane involves the CFTR Cl channel; however, conductive pathways for Cl movement across the basolateral membrane have been little studied. Here, we determined the regulation and single-channel properties of the Cl conductance (GCl) in airway surface epithelia using epithelial cultures from human or bovine trachea and freshly isolated ciliated cells from the human nasal epithelium. In Ussing chamber studies, a swellingactivated basolateral GCl was found, which was further stimulated by forskolin and blocked by N-phenylanthranilic acid (DPC) = sucrose > flufenamic acid = niflumic acid = glibenclamide > CdCl2 = 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) = DIDS = ZnCl2 > tamoxifen > 4,4′-dinitro-2,2′-stilbene- disulfonate disodium salt (DNDS). In whole cell patch-clamp experiments, three types of GCl were identified: 1) a voltage-activated, DIDS- (but not Cd-) blockable and osmosensitive GCl; 2) an inwardly rectifying, hyperpolarization-activated and Cd-sensitive GCl; and 3) a forskolin-activated, linear GCl, which was insensitive to Cd and DIDS. In cell-attached patch-clamp recordings, the basolateral pole of isolated ciliated cells expressed three types of Cl channels: 1) an outwardly rectifying, swelling-activated Cl channel; 2) a strongly inwardly rectifying Cl channel; and 3) a forskolin-activated, low-conductance channel. We propose that, depending on the driving force for Cl across the apical membrane, basolateral Cl channels confine Cl- secretion or support transcellular Cl- absorption.
| Original language | English (US) |
|---|---|
| Journal | American Journal of Physiology - Lung Cellular and Molecular Physiology |
| Volume | 292 |
| Issue number | 6 |
| DOIs | |
| State | Published - Jun 2007 |
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Keywords
- Absorption
- Airway epithelium
- Basolateral membrane
- Chloride channels
- Ion transport
- Secretion
ASJC Scopus subject areas
- Pulmonary and Respiratory Medicine
- Cell Biology
- Physiology
Cite this
Basolateral Cl channels in primary airway epithelial cultures. / Fischer, Horst; Illek, Beate; Finkbeiner, Walter E.; Widdicombe, Jonathan.
In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 292, No. 6, 06.2007.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Basolateral Cl channels in primary airway epithelial cultures
AU - Fischer, Horst
AU - Illek, Beate
AU - Finkbeiner, Walter E.
AU - Widdicombe, Jonathan
PY - 2007/6
Y1 - 2007/6
N2 - Salt and water absorption and secretion across the airway epithelium are important for maintaining the thin film of liquid lining the surface of the airway epithelium. Movement of Cl across the apical membrane involves the CFTR Cl channel; however, conductive pathways for Cl movement across the basolateral membrane have been little studied. Here, we determined the regulation and single-channel properties of the Cl conductance (GCl) in airway surface epithelia using epithelial cultures from human or bovine trachea and freshly isolated ciliated cells from the human nasal epithelium. In Ussing chamber studies, a swellingactivated basolateral GCl was found, which was further stimulated by forskolin and blocked by N-phenylanthranilic acid (DPC) = sucrose > flufenamic acid = niflumic acid = glibenclamide > CdCl2 = 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) = DIDS = ZnCl2 > tamoxifen > 4,4′-dinitro-2,2′-stilbene- disulfonate disodium salt (DNDS). In whole cell patch-clamp experiments, three types of GCl were identified: 1) a voltage-activated, DIDS- (but not Cd-) blockable and osmosensitive GCl; 2) an inwardly rectifying, hyperpolarization-activated and Cd-sensitive GCl; and 3) a forskolin-activated, linear GCl, which was insensitive to Cd and DIDS. In cell-attached patch-clamp recordings, the basolateral pole of isolated ciliated cells expressed three types of Cl channels: 1) an outwardly rectifying, swelling-activated Cl channel; 2) a strongly inwardly rectifying Cl channel; and 3) a forskolin-activated, low-conductance channel. We propose that, depending on the driving force for Cl across the apical membrane, basolateral Cl channels confine Cl- secretion or support transcellular Cl- absorption.
AB - Salt and water absorption and secretion across the airway epithelium are important for maintaining the thin film of liquid lining the surface of the airway epithelium. Movement of Cl across the apical membrane involves the CFTR Cl channel; however, conductive pathways for Cl movement across the basolateral membrane have been little studied. Here, we determined the regulation and single-channel properties of the Cl conductance (GCl) in airway surface epithelia using epithelial cultures from human or bovine trachea and freshly isolated ciliated cells from the human nasal epithelium. In Ussing chamber studies, a swellingactivated basolateral GCl was found, which was further stimulated by forskolin and blocked by N-phenylanthranilic acid (DPC) = sucrose > flufenamic acid = niflumic acid = glibenclamide > CdCl2 = 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) = DIDS = ZnCl2 > tamoxifen > 4,4′-dinitro-2,2′-stilbene- disulfonate disodium salt (DNDS). In whole cell patch-clamp experiments, three types of GCl were identified: 1) a voltage-activated, DIDS- (but not Cd-) blockable and osmosensitive GCl; 2) an inwardly rectifying, hyperpolarization-activated and Cd-sensitive GCl; and 3) a forskolin-activated, linear GCl, which was insensitive to Cd and DIDS. In cell-attached patch-clamp recordings, the basolateral pole of isolated ciliated cells expressed three types of Cl channels: 1) an outwardly rectifying, swelling-activated Cl channel; 2) a strongly inwardly rectifying Cl channel; and 3) a forskolin-activated, low-conductance channel. We propose that, depending on the driving force for Cl across the apical membrane, basolateral Cl channels confine Cl- secretion or support transcellular Cl- absorption.
KW - Absorption
KW - Airway epithelium
KW - Basolateral membrane
KW - Chloride channels
KW - Ion transport
KW - Secretion
UR - http://www.scopus.com/inward/record.url?scp=34447553334&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34447553334&partnerID=8YFLogxK
U2 - 10.1152/ajplung.00032.2007
DO - 10.1152/ajplung.00032.2007
M3 - Article
C2 - 17322286
AN - SCOPUS:34447553334
VL - 292
JO - American Journal of Physiology - Renal Fluid and Electrolyte Physiology
JF - American Journal of Physiology - Renal Fluid and Electrolyte Physiology
SN - 1931-857X
IS - 6
ER -