Adam, Gabriele and Ousingsawat, Jiraporn and Schreiber, Rainer and Kunzelmann, Karl (2005) Increase in intracellular Cl- concentration by cAMP- and Ca2+-dependent stimulation of M1 collecting duct cells. PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 449 (5). pp. 470-478. ISSN 0031-6768, 1432-2013
Full text not available from this repository. (Request a copy)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 muM) and forskolin (2 muM) increased [Cl-](i) by 9.6 +/- 1.5 mM (n = 35). Similarly, ATP (100 muM) increased [Cl-](i) transiently by 9.5 +/- 2.2 mM (n = 17). The increase in [CF]i was reduced by the Na+/K+/2 Cl--cortransporter-1 (NKCCl) blocker azosemide (100[muM), the CFTR blocker SP-303 (50 muM), the blocker of Ca2+-activated Cl- channels DIDS (100 muM) or the ENaC blocker amiloride (10 muM). 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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | TRANSMEMBRANE CONDUCTANCE REGULATOR; GREEN FLUORESCENT PROTEIN; EPITHELIAL NA+ CONDUCTANCE; CYSTIC-FIBROSIS AIRWAYS; XENOPUS OOCYTES; EXTRACELLULAR NUCLEOTIDES; PURINERGIC STIMULATION; SODIUM-CHANNELS; HUMAN COLON; CFTR; CFTR; ENaC; intracellular chloride; purinergic stimulation; cystic fibrosis; NKCC1; YFP; Cl- channels; epithelial transport |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Physiologie > Prof. Dr. Karl Kunzelmann |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 31 May 2021 07:14 |
| Last Modified: | 31 May 2021 07:14 |
| URI: | https://pred.uni-regensburg.de/id/eprint/36560 |
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