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J Biol Chem, Vol. 274, Issue 21, 14670-14677, May 21, 1999
/HCO3
Exchange in Mouse Submandibular and Pancreatic Ducts
,
From the We have demonstrated previously the regulation of
Cl
Department of Pharmacology, Yonsei
University College of Medicine, Seoul 120-752, Korea, and the
¶ Department of Physiology and the Program in Molecular
Biophysics, University of Texas Southwestern Medical Center,
Dallas, Texas 75235
/HCO3
exchange
activity by the cystic fibrosis transmembrane conductance regulator
(CFTR) in model systems of cells stably or transiently transfected with
CFTR (Lee, M. G., Wigley, W. C., Zeng, W., Noel, L. E.,
Marino, C. R., Thomas, P. J., and Muallem, S. (1999)
J. Biol. Chem. 274, 3414-3421). In the present work
we examine the significance of this regulation in cells naturally
expressing CFTR. These include the human colonic T84 cell line and the
mouse submandibular gland and pancreatic ducts, tissues that express high levels of CFTR in the luminal membrane. As in heterologous expression systems, stimulation of T84 cells with forskolin increased the Cl
/HCO3
exchange
activity independently of CFTR Cl
channel activity.
Freshly isolated submandibular gland ducts from wild type mice showed
variable Cl
/HCO3
exchange activity. Measurement of
[Cl
]i revealed that this was
largely the result of variable steady-state
[Cl
]i. Membrane depolarization
with 5 mM Ba2+ or 100 mM
K+ increased and stabilized
[Cl
]i. Under depolarized
conditions wild type and
F/
F mice had comparable basal
Cl
/HCO3
exchange
activity. Notably, stimulation with forskolin increased Cl
/HCO3
exchange
activity in submandibular gland ducts from wild type but not
F/
F
mice. Microperfusion of the main pancreatic duct showed
Cl
/HCO3
exchange
activity in both the basolateral and luminal membranes. Stimulation of
ducts from wild type animals with forskolin had no effect on
basolateral but markedly stimulated luminal
Cl
/HCO3
exchange
activity. By contrast, forskolin had no effect on either basolateral or
luminal Cl
/HCO3
exchange activity of ducts from
F/
F animals. We conclude that CFTR regulates luminal
Cl
/HCO3
exchange
activity in CFTR-expressing cells, and we discuss the possible
physiological significance of these findings regarding cystic fibrosis.
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