Activation of CFTR Cl- conductance in polarized T84 cells by luminal extracellular ATP.
Luminal extracellular ATP evoked a bumetanide-sensitive short-circuit current in cultured T84 cell epithelia (90.2 +/- 18.2 microA/cm2 at 100 microM ATP, apparent 50% effective concentration, 11.5 microM). ATP appeared to increase the Cl- conductance of the apical membrane but not the driving force for Cl- secretion determined by basolateral membrane K+ conductance. Specifically, the magnitude of Cl- secretion stimulated by ATP was independent of basal current, and forskolin pretreatment abolished subsequent stimulation of Cl- secretion by ATP. Whereas ATP stimulated modest production of adenosine 3',5'-cyclic monophosphate (cAMP) by T84 cells, ATP caused smaller increases in intracellular Ca2+ and inositol phosphate activities than the Ca(2+)-signaling Cl- secretagogue carbachol. An inhibitor of 5'-nucleotidase, alpha,beta-methyleneadenosine 5'-diphosphate, blocked most of the response to luminal ATP. The adenosine receptor antagonist 8-(p-sulfophenyl)theophylline blocked both the luminal ATP-dependent generation of cAMP and Cl- secretion when administered to the luminal but not submucosal bath. These results demonstrate that the Cl- secretion stimulated by luminal ATP is mediated by a A2-adenosine receptor located on the apical cell membrane. Thus metabolism of extracellular ATP to adenosine regulates the activity of cystic fibrosis transmembrane conductor regulator (CFTR) in the apical membrane of polarized T84 cells.
Stutts
M J
MJ
Department of Medicine, University of North Carolina at Chapel Hill 27599-7020.
Lazarowski
E R
ER
Paradiso
A M
AM
Boucher
R C
RC
D000255:Adenosine Triphosphate / Q000494:pharmacology / Q000502:physiology*; D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D016764:Cell Polarity; D018118:Chloride Channels / Q000502:physiology; D002712:Chlorides / Q000502:physiology*; D005576:Colforsin / Q000494:pharmacology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004553:Electric Conductivity; D005110:Extracellular Space / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism*; D008565:Membrane Proteins / Q000502:physiology*; D018047:Receptors, Purinergic P1 / Q000502:physiology; D013806:Theophylline / Q000031:analogs & derivatives / Q000494:pharmacology
D018118:Chloride Channels; D002712:Chlorides; D008565:Membrane Proteins; D018047:Receptors, Purinergic P1; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; C031433:8-(4-sulfophenyl)theophylline; D000255:Adenosine Triphosphate; D002217:Carbachol; D013806:Theophylline