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Published ahead of print on August 28, 2003, doi:10.1165/rcmb.2003-0184OC

Am. J. Respir. Cell Mol. Biol., Volume 30, Number 3, March 2004, 388-395

A more recent version of this article appeared on March 1, 2004
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Submitted on May 7, 2003
Revised on August 28, 2003

ATP Release Triggered by Activation of the Ca2+ -activated K+ Channel in Human Airway Calu-3 Cells

Yasushi Ito1*, Masami Son1, Shinji Sato1, Takayuki Ishikawa1, Masashi Kondo1, Shinsuke Nakayama2, Kaoru Shimokata1, and Hiroaki Kume1

1 Division of Respiratory Diseases, Department of Internal Medicine, Nagoya University Graduate School of Medicine, Nagoya, Aichi, Japan, 2 Department of Cellular Physiology, Nagoya University Graduate School of Medicine, Nagoya, Aichi, Japan

* To whom correspondence should be addressed. E-mail: itoyasu{at}med.nagoya-u.ac.jp.

Airway mucociliary clearance is subject to the autocrine/paracrine regulation of extracellular nucleotides released from the airway epithelial cells. The present study was performed in pursuit of effective modulators of ATP release under physiological conditions in polarized human airway epithelial cells (Calu-3). Neither isoproterenol, forskolin, nor ionomycin augmented extracellular ATP release detected by luciferase assay. However, direct activation of the human intermediated conductance, Ca2+-activated K+ channel (hIK-1) by 1-ethyl-2-benzimdazolinone (1-EBIO, 1 mM) and chlorzoxazone (CZ, 1 mM) did predominantly in the apical compartment. Measurement of fluo-3 signals revealed that 1-EBIO- and CZ-stimulated cytosolic Ca2+ mobilization was suppressed by the presence of MRS-2179, a specific P2Y1 receptor antagonist. The hIK-1-mediated ATP release was inhibited by a hIK-1 blocker (charybdotoxin), and a Na+-K+-2Cl- cotransport blocker (bumetanide) without interruption by GdCl3, an inhibitor of stretch-activated non-selective cation (SA) channels, or glybenclamide, a blocker of the cystic fibrosis transmembrane conductance regulator (CFTR). These results suggest that a cell volume decrease via the hIK-1-mediated KCl loss and the resultant induction of a regulatory volume increase via the Na+-K+-2Cl- transporter may trigger release of ATP, which causes P2Y1-mediated Ca2+ mobilization, through mechanisms unrelated to the CFTR and SA channels.




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