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and HCO
Departments of 1 Anesthesiology, 2 Pediatrics, 3 Environmental Health Sciences, and 4 Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, Alabama 35233
We
isolated and cultured fetal distal lung epithelial (FDLE) cells from
17- to 19-day rat fetuses and assayed for anion secretion in
Ussing chambers. With symmetrical Ringer solutions, basal short-circuit currents (Isc) and transepithelial resistances
were 7.9 ± 0.5 µA/cm2 and 1,018 ± 73 
· cm2, respectively (means ± 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 µM) decreased
forskolin-stimulated Isc from 7.51 ± 0.2 µA/cm2 to 5.62 ± 0.53, whereas basolateral
4,4'-dinitrostilbene-2,2'-disulfonate (5 mM), an inhibitor of
HCO
- or
HCO-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. Immunolocalization
studies were consistent with the expression of cystic fibrosis
transmembrane conductance regulator (CFTR) protein in FDLE cells. In
aggregate, these findings indicate the presence of cAMP-activated
Cl and HCO
short-circuit current; amiloride; nystatin; swelling-activated conductance; immunocytochemistry; adenosine 3',5'-cyclic monophosphate
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