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Articles in PresS, published online ahead of print December 7, 2001
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00370.2001
Submitted on September 19, 2001
Accepted on November 27, 2001
1 Anesthesiology, University of Alabama, Birmingham, AL, USA
2 Pediatrics, University of Alabama, Birmingham, AL, USA
3 Environmental Health Sciences, University of Alabama, Birmingham, AL, USA
4 Anesthesiology, University of Alabama, Birmingham, AL, USA; Pediatrics, University of Alabama, Birmingham, AL, USA; Environmental Health Sciences, University of Alabama, Birmingham, AL, USA
* To whom correspondence should be addressed. E-mail: sadis.matalon{at}ccc.uab.edu.
We isolated and cultured fetal distal lung epithelial (FDLE) cells from 19 d rat fetuses and assayed for anion secretion in Ussing chambers. With symmetric Ringer's solutions, basal short-circuit currents (Isc) and transepithelial resistances were 7.9±0.5 µA/cm2 and 1018±73 Ohm*cm2 (mean±SE; n=12). Apical amiloride (10 µM) inhibited basal Isc by ~50%. Subsequent addition of forskolin (10 µM) increased Isc from 3.9±0.63 µA/cm2 to 7.51±0.2 µA/cm2 (n=12). Basolateral bumetanide (100 mM) decreased forskolin-stimulated Isc from 7.51±0.2 µA/cm2 to 5.62±0.53 while basolateral DNDS (5 mM), an inhibitor of the Na-HCO3- exchanger, blocked the remaining Isc. Forskolin addition evoked currents of similar fractional magnitudes in symmetric Cl- or HCO3- free solutions; however, no response was seen using HCO3- and Cl- free solutions. The forskolin-stimulated Isc was inhibited by glibenclamide but not apical DIDS. Glibenclamide also blocked forskolin-induced Isc across monolayers having nystatin-permeablized basolateral membranes. Finally, CFTR protein was immunolocalized in the apical membranes. In aggregate, these findings indicate the presence of cAMP-activated Cl- and HCO3-secretion across rat FDLE cells mediated via CFTR.
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