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Am. J. Respir. Cell Mol. Biol., Volume 25, Number 1, July, 2001 14-20

Liquid and Ion Transport by Fetal Airway and Lung Epithelia of Mice Deficient in Sodium-Potassium-2-Chloride Transporter

Daniel J. Gillie, Amy J. Pace, Ray J. Coakley, Beverly H. Koller, and Pierre M. Barker

Departments of Pediatrics and Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina

Chloride (Cl-) movement across fetal lung epithelia is thought to be mediated by the sodium-potassium-2-Cl- cotransporter NKCC1. We studied the role of NKCC1 in Cl- and liquid secretion in late-gestation NKCC-null (-/-) and littermate control fetal mouse lung. NKCC -/- mice had decreased lung water compared with littermate controls (wet/dry: control, 8.01 ± 0.09; NKCC -/-, 7.06 ± 0.14). Liquid secretion by 17-d NKCC -/- distal lung explants was similar to control explants. Bumetanide inhibited basal liquid secretion in control but not NKCC -/- explants (expansion over 48 h: control, 35 ± 4%; NKCC -/- 46 ± 7%). Treatment with 4,4'-diisothiocyanto-stilbene-2,2'-disulfonic acid (DIDS) decreased liquid secretion in both control and NKCC -/- explants. Basal transepithelial potential difference (PD) of control tracheal explants was higher than that of NKCC -/- (control, -13.7 ± 0.5 mV; NKCC -/-, -11.6 ± 0.6 mV). Amiloride (10-4 M) inhibited basal PD to the same extent in control and NKCC -/- mice. Terbutaline-stimulated hyperpolarization was less in NKCC -/- than in control tracheas (Delta PD: control, -10.8 ± 1.33 mV; NKCC -/-, -6.1 ± 0.7 mV) and was inhibited by DIDS and acetazolamide in NKCC -/- but not wild-type explants. We conclude that NKCC is rate-limiting for transcellular Cl- transport, and that alternative anion transport mechanisms can sustain liquid production at near-normal levels in the fetal NKCC -/- mouse lung.

Abbreviations: anion exchanger, AE; cyclic adenosine monophosphate, cAMP; chloride, Cl-; 4,4'-diisothiocyanto-stilbene-2,2'-disulfonic acid, DIDS; Na+/H+ exchanger, NHE; sodium-potassium-2-chloride cotransporter, NKCC; potential difference, PD.




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