TITLE

Increase in intracellular Cl- concentration by cAMP- and Ca2+-dependent stimulation of M1 collecting duct cells

AUTHOR(S)
Adam, Gabriele; Ousingsawat, Jiraporn; Schreiber, Rainer; Kunzelmann, Karl
PUB. DATE
February 2005
SOURCE
Pflugers Archiv European Journal of Physiology;Feb2005, Vol. 449 Issue 5, p470
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
In the lungs of cystic fibrosis (CF) patients, mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) lead to defective Cl- secretion and hyperabsorption of electrolytes. This may be a an important cause for the defective mucociliary clearance in CF lungs. Previous studies have suggested that inhibition of ENaC during activation of CFTR or by purinergic stimulation could be related to an increase in the intracellular [Cl-]i. This was examined in the present study using cultured mouse M1 collecting duct cells transfected with the chloride-sensitive enhanced yellow fluorescent protein YFPV163S. Calibration experiments showed a linear decrease of YFP fluorescence intensity with increasing [Cl-]i (0-100 mM). Activation of CFTR by isobutyl-1-methylxanthine (IBMX, 100 µM) and forskolin (2 µM) increased [Cl-]i by 9.6±1.5 mM (n=35). Similarly, ATP (100 µM) increased [Cl-]i transiently by 9.5±2.2 mM (n=17). The increase in [Cl-]i was reduced by the Na+/K+/2 Cl--cortransporter-1 (NKCC1) blocker azosemide (100 µM), the CFTR blocker SP-303 (50 µM), the blocker of Ca2+-activated Cl- channels DIDS (100 µM) or the ENaC blocker amiloride (10 µM). Changes in YFPV163S fluorescence were not due to changes in cell volume or intracellular pH. The present data thus demonstrate an increase in [Cl-]i following stimulation with secretagogues, which could participate in the inhibition of ENaC.
ACCESSION #
15909705

 

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