title: Up-regulation of A 2B adenosine receptor in A 2A adenosine receptor knockout mouse coronary artery.
abstract: In this study, we looked into possible compensatory changes of other adenosine receptors (ARs) in A(2A) genetic knockout mice (A2AKO) as well as the functional role of nitric oxide (NO) in A(2A) AR-mediated vasodilation. Gene expression of ARs from coronary arteries of A(2A) AR wild type mice (A2AWT) and A2AKO was studied using real-time PCR. Functional studies were carried out in isolated heart and isolated coronary artery preparations. A(2B) AR was found to be 4.5 fold higher in A2AKO than in A2AWT, while A(2A) AR expression was absent in A2AKO. There was no difference in A(1) and A(3) ARs between WT and KO animals. The concentration-relaxation curve for adenosine-5'-N-ethylcarboxamide (NECA, non-selective AR agonist) in isolated coronary arterial rings in A2AKO was shifted to the left when compared to A2AWT. The concentration-response curve for A(2B) selective agonist (BAY 60-6583) was also shifted to the left in A2AKO hearts. L-NAME, a non-specific NO synthase inhibitor, did not affect baseline coronary flow (CF) until the concentration reached 10 microM in A2AWT (76.32+/-11.35% from baseline, n=5). In A2AKO, the CF decreased significantly by L-NAME only at a higher concentration (100 microM, 93.32+/-5.8% from baseline, n=5). L-NMA (1 microM, n=4), another non-specific NO synthase inhibitor, also demonstrated similar results in decreasing CF (59.66+/-3.23% from baseline in A2AWT, while 81.76+/-8.91% in A2AKO). It was further demonstrated that the increase in CF by 100 microM NECA was significantly blunted with 10 microM L-NAME (377.08+/-25.23% to 305.41+/-30.73%, n=9) in A2AWT but not in A2AKO (153.66+/-22.7% to 143.88+/-36.65%, n=5). Similar results were also found using 50 nM of CGS-21680 instead of NECA in A2AWT (346+/-22.85 to 277+/-31.39, n=6). No change in CF to CGS-21680 was noted in A(2A)AKO. Our data demonstrate, for the first time, that coronary A(2B) AR was up-regulated in mice deficient in A(2A) AR. We also provide direct evidence supporting a role for NO in A(2A) AR-mediated coronary vasodilation. The data further support the role for A(2A) AR in the regulation of basal coronary tone through the release of NO.
authors.affiliation: Department of Physiology and Pharmacology, West Virginia University Health Sciences Center, Morgantown, WV 26506-9104, USA.
authors.forename: Bunyen
authors.initials: B
authors.lastname: Teng
authors.forename: Catherine
authors.initials: C
authors.lastname: Ledent
authors.forename: S Jamal
authors.initials: SJ
authors.lastname: Mustafa
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D058908:Adenosine A2 Receptor Agonists; D019830:Adenosine-5'-(N-ethylcarboxamide) / Q000494:pharmacology; D000631:Aminopyridines / Q000494:pharmacology; D000818:Animals; D003326:Coronary Circulation / Q000187:drug effects; D003331:Coronary Vessels / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006339:Heart Rate / Q000187:drug effects; D066298:In Vitro Techniques; D051379:Mice; D018345:Mice, Knockout; D009213:Myography; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D010477:Perfusion; D010627:Phenethylamines / Q000494:pharmacology; D043705:Receptor, Adenosine A2A / Q000172:deficiency*; D043704:Receptor, Adenosine A2B / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics*; D014664:Vasodilation / Q000187:drug effects
chemical_list: D058908:Adenosine A2 Receptor Agonists; D000631:Aminopyridines; C518875:BAY 60-6583; D010627:Phenethylamines; D043705:Receptor, Adenosine A2A; D043704:Receptor, Adenosine A2B; C061282:2-(4-(2-carboxyethyl)phenethylamino)-5'-N-ethylcarboxamidoadenosine; D009569:Nitric Oxide; D019830:Adenosine-5'-(N-ethylcarboxamide); D052250:Nitric Oxide Synthase Type III; D000241:Adenosine; D019331:NG-Nitroarginine Methyl Ester