TY - JOUR T1 - Canine Mast Cell Adenosine Receptors: Cloning and Expression of the A<sub>3</sub> Receptor and Evidence that Degranulation Is Mediated by the A<sub>2B</sub> Receptor JF - Molecular Pharmacology JO - Mol Pharmacol SP - 846 LP - 860 DO - 10.1124/mol.52.5.846 VL - 52 IS - 5 AU - John A. Auchampach AU - Xiaowei Jin AU - Tina C. Wan AU - George H. Caughey AU - Joel Linden Y1 - 1997/11/01 UR - http://molpharm.aspetjournals.org/content/52/5/846.abstract N2 - We cloned and characterized the canine A3 adenosine receptor (AR) and examined AR-induced degranulation of the BR line of canine mastocytoma cells. Canine A3AR transcript is found predominantly in spleen, lung, liver, and testes and encodes a 314-amino acid heptahelical receptor.125I-N 6-Aminobenzyladenosine binds to two affinity states of canine A3AR withK D values of 0.7 ± 0.1 and 16 ± 0.8 nm, reflecting G protein-coupled and -uncoupled receptors, respectively. Xanthine antagonists bind with similar affinities to human, canine, and rabbit receptors but with 80–400-fold lower affinities to rat A3AR. Although canine BR mastocytoma cells contain A1AR, A2BAR, and A3AR, degranulation seems to be mediated primarily by A2BARs stimulated by the nonselective agonist 5′-N-ethylcarboxamidoadenosine (NECA) but not by the A3-selective agonistN 6-(3-iodobenzyl)adenosine-5′-N-methylcarboxamide. NECA-stimulated degranulation is not prevented by pertussis toxin and is blocked by enprofylline (K i = 7 μm), an antiasthmatic xanthine with low affinity (K i &gt; 100 μm) for A1AR, A2AAR, and A3AR. NECA increases canine mastocytoma cell cAMP, Ca2+, and inositol trisphosphate levels; these responses are antagonized half-maximally by 7–15 μm enprofylline. The results suggest that (i) the cloned canine A3AR is structurally and pharmacologically more similar to human than to rat A3AR; (ii) the A2BAR, and not the A1AR or A3AR, is principally responsible for adenosine-mediated degranulation of canine BR mastocytoma cells; and (iii) the BR cell A2BAR couples to both Ca2+ mobilization and cAMP accumulation. Although A2B receptors play a major role in the regulation of BR mast cell degranulation, multiple AR subtypes and G proteins may influence mast cell functions. ER -