Published ahead of print on May 18, 2006, doi:10.1165/rcmb.2005-0290OC
© 2006 American Thoracic Society DOI: 10.1165/rcmb.2005-0290OC Adenosine A1 Receptors Mediate Mobilization of Calcium in Human Bronchial Smooth Muscle CellsDepartment of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts Correspondence and requests for reprints should be addressed to Michael F. Ethier, Department of Medicine, 364 Plantation Street, LRB, Room 370A, University of Massachusetts Medical School, Worcester, MA 01605. E-mail: Michael.ethier{at}umassmed.edu Adenosine stimulates contraction of airway smooth muscle, but the mechanism is widely considered indirect, depending on release of contractile agonists from mast cells and nerves. The goal was to determine whether adenosine, by itself, directly regulates calcium signaling in human bronchial smooth muscle cells (HBSMC). Primary cultures of HBSMC from normal subjects were loaded with fura 2-AM, and cytosolic calcium concentrations ([Ca2+]i) were determined ratiometrically by imaging single cells. The nonselective adenosine receptor agonist, 5'-N-ethylcarboxamidoadenosine (NECA), and the adenosine A1 receptor agonist, N6-cyclopentyladenosine (CPA), both stimulated rapid, transient increases in [Ca2+]i. In contrast, there were no calcium responses to 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxamido-adenosine (100 nM) or N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide (100 nM), selective agonists at adenosine A2A receptors and adenosine A3 receptors, respectively. Calcium responses to NECA and CPA were inhibited by 8-cyclopentyl-1,3-dipropylxanthine, an adenosine A1 receptor antagonist, and by pertussis toxin (PTX). In other experiments, NECA stimulated calcium transients in the absence of extracellular calcium, but not when cells were preincubated in cyclopiazonic acid or thapsigargin to empty intracellular calcium stores. Calcium responses were attenuated by xestospongin C and 2-aminoethoxydiphenylborane, inhibitors of inositol trisphosphate (IP3) receptors, and by U73122, an inhibitor of phospholipase C. It was concluded that stimulation of adenosine A1 receptors on HBSMC rapidly mobilizes intracellular calcium stores by a mechanism dependent on PTX-sensitive G proteins, and IP3 signaling. These findings suggest that, in addition to its well-established indirect effects on HBSMC, adenosine also has direct effects on contractile signaling pathways.
Key Words: adenosine A1 receptor • calcium • cAMP • human bronchial smooth muscle • insulin This article has been cited by other articles:
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