Characterization and comparison of ion transport across sheep and human airway epithelium.
Diana M. Steel, Alexander D. Graham, Geddes Dm, Eric W.F.W. Alton
Abstract
Diana M. Steel, Alexander D. Graham, Geddes Dm, Eric W.F.W. Alton
Abstract
This study aimed to assess the suitability of sheep tracheal epithelium as a model for studies of human airway ion transport. Ovine and human airway epithelium were mounted in Ussing chambers under short circuit conditions. Bumetanide (100 microM) reduced short-circuit current (Isc) by a mean of 21.3% +/- SEM 2.0, n = 8, in sheep, and 30.4% +/- 9.7, n = 3, in human airway epithelium. Acetazolamide (100 microM) decreased Isc by 10.6% +/- 1.2, n = 18, in sheep, and 5.8% +/- 2.9, n = 3, in human airways. Phloridzin (200 microM) reduced Isc by 4.7% +/- 0.8, n = 7, and 3.1% +/- 5.1, n = 3 in sheep and human tissue respectively. Amiloride (100 microM) decreased Isc by 42.9% +/- 3.5, n = 12, in sheep airways, whilst bathing the mucosal surface with Na(+)-free solutions reduced Isc by 67.4% +/- 4.2, n = 18. The sequential addition of acetazolamide, bumetanide, phloridzin, amiloride and mucosal Na(+)-free solutions totally inhibited the basal Isc in both sheep and human tissues, suggesting that Cl- and HCO3- secretion, Na(+)-glucose co-transport and amiloride-sensitive and -insensitive Na+ absorption contribute to the Isc. The similarities between the species suggest that sheep tracheal epithelium is a useful model for basal studies of airway ion transport, and may prove a valuable tool for further regulatory studies.
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This study aimed to assess the suitability of sheep tracheal epithelium as a model for studies of human airway ion transport. Ovine and human airway epithelium were mounted in Ussing chambers under short circuit conditions. Bumetanide (100 microM) reduced short-circuit current (Isc) by a mean of 21.3% +/- SEM 2.0, n = 8, in sheep, and 30.4% +/- 9.7, n = 3, in human airway epithelium. Acetazolamide (100 microM) decreased Isc by 10.6% +/- 1.2, n = 18, in sheep, and 5.8% +/- 2.9, n = 3, in human airways. Phloridzin (200 microM) reduced Isc by 4.7% +/- 0.8, n = 7, and 3.1% +/- 5.1, n = 3 in sheep and human tissue respectively. Amiloride (100 microM) decreased Isc by 42.9% +/- 3.5, n = 12, in sheep airways, whilst bathing the mucosal surface with Na(+)-free solutions reduced Isc by 67.4% +/- 4.2, n = 18. The sequential addition of acetazolamide, bumetanide, phloridzin, amiloride and mucosal Na(+)-free solutions totally inhibited the basal Isc in both sheep and human tissues, suggesting that Cl- and HCO3- secretion, Na(+)-glucose co-transport and amiloride-sensitive and -insensitive Na+ absorption contribute to the Isc. The similarities between the species suggest that sheep tracheal epithelium is a useful model for basal studies of airway ion transport, and may prove a valuable tool for further regulatory studies.
Key concepts: Amiloride, Bumetanide, Respiratory epithelium, Epithelium, Ion transporter, Acetazolamide, Transepithelial potential difference, Chemistry