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Am. J. Respir. Cell Mol. Biol., Volume 25, Number 3, September, 2001 370-376

Differential Regulations between Adenosine Triphosphate (ATP)- and Uridine Triphosphate-Induced Clminus Secretion in Bovine Tracheal Epithelium
Direct Stimulation of P1-like Receptor by ATP

Soichiro Kanoh, Mitsuko Kondo, Jun Tamaoki, Hideo Kobayashi, Kazuo Motoyoshi, and Atsushi Nagai

Third Department of Medicine, National Defense Medical College, Saitama; and First Department of Medicine, Tokyo Women's Medical University School of Medicine, Tokyo, Japan

Adenosine 5'-triphosphate (ATP) stimulates airway epithelial Cl- secretion in a complicated manner. We examined the difference between ATP- and uridine 5'-triphosphate (UTP)-induced responses of short-circuit current (Isc) in bovine tracheal epithelium treated with amiloride. Each nucleotide caused an increase in Isc composed of the first and second peaks, where the second peak induced by ATP was higher compared with UTP. The ATP-induced second peak was inhibited by the protein kinase (PK) A inhibitor H89, saturation of P1 receptor with adenosine, and the P1 receptor antagonist 8-p-sulfophenyltheophylline, but not by the Ca2+ chelator ethyleneglycol-bis-(beta -aminoethyl ether)-N,N,N',N'-tetraacetic acid plus the endoplasmic reticulum Ca2+-pump inhibitor thapsigargin, the adenosine breakdown enzyme adenosine deaminase, the ectonucleotidase inhibitor alpha ,beta -methyleneadenosine 5'-diphosphate, or saturation of P2Y2 receptor with UTP. Thus, the response is associated with PKA-dependent pathway via P1-like receptor but not with Ca2+-dependent pathway via P2Y2 receptor, and ATP degradation products do not contribute to this response. Further, stimulation of cells with ATP increased PKA activity. In addition, pretreatment with glybenclamide, an inhibitor of cystic fibrosis transmembrane conductance regulator, reduced the second peak of Isc induced by ATP but was without effect on that induced by UTP. Therefore, ATP stimulates glybenclamide-sensitive Cl- secretion, and this action is partly mediated by PKA-dependent pathway via P1-like receptor.

Abbreviations: adenosine deaminase, ADA; adenosine, ADO; alpha ,beta -methyleneadenosine 5'-diphosphate, AMP-CP; adenosine 5'-triphosphate, ATP; intracellular Ca2+ concentration, [Ca2+]i; cyclic adenosine 3',5'-monophosphate, cAMP; cystic fibrosis transmembrane conductance regulator, CFTR; 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, DIDS; diphenylamine-2-carboxylate, DPC; ethyleneglyco-bis-(beta -amino-ethyl ether)-N,N, N',N',-tetraacetic acid, EGTA; short-circuit current, Isc; alpha ,beta -methylene-ATP, alpha ,beta -MeATP; 2-methylthio-ATP, 2-MeSATP; outwardly rectifying Cl- channel, ORCC; phosphate-buffered saline, PBS; potential difference, PD; protein kinase, PK; phospholipase, PL; 8-p-sulfophenyltheophylline, 8-SPT; standard error, SE; thapsigargin, TG; uridine 5'-trisphosphate, UTP.




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Copyright © 2001 American Thoracic Society.