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Am J Physiol Heart Circ Physiol 279: H1397-H1410, 2000;
0363-6135/00 $5.00
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Vol. 279, Issue 3, H1397-H1410, September 2000

Selective transport of adenosine into porcine coronary smooth muscle

L. J. Rubin1,3, L. R. Johnson1, J. R. Dodam1, A. K. Dhalla1, L. Magliola2, M. H. Laughlin1,2,3, and A. W. Jones2,3

1 Department of Veterinary Biomedical Sciences and 2 Department of Medical Physiology, 3 Dalton Cardiovascular Research Center, University of Missouri-Columbia, Columbia, Missouri 65211

Adenosine (ADO), an endogenous regulator of coronary vascular tone, enhances vasorelaxation in the presence of nucleoside transport inhibitors such as dipyridamole. We tested the hypothesis that coronary smooth muscle (CSM) contains a high-affinity transporter for ADO. ADO-mediated relaxation of isolated large and small porcine coronary artery rings was enhanced 12-fold and 3.4-fold, respectively, by the transport inhibitor, S-(4-nitrobenzyl)-6-thioinosine (NBTI). Enhanced relaxation was independent of endothelium and was selective for ADO over synthetic analogs. Uptake of [3H]ADO into freshly dissociated CSM cells or endothelium-denuded rings was linear and concentration dependent. Kinetic analysis yielded a maximum uptake (Vmax) of 67 ± 7.0 pmol · mg protein-1 · min-1 and a Michaelis constant (Km) of 10.5 ± 5.8 µM in isolated cells and a Vmax of 5.1 ± 0.5 pmol · min-1 · mg wet wt-1 and a Km of 17.6 ± 2.6 µM in intact rings. NBTI inhibited transport into small arteries (IC50 = 42 nM) and cells. Analyses of extracellular space and diffusion kinetics using [3H]sucrose indicate the Vmax and Km for ADO transport are sufficient to clear a significant amount of extracellular adenosine. These data indicate CSM possess a high-affinity nucleoside transporter and that the activity of this transporter is sufficient to modulate ADO sensitivity of large and small coronary arteries.

dipyridamole; S-(4-nitrobenzyl)-6-thioinosine; erythro-9-(2-hydroxy-3-nonyl)-adenine hydrochloride; 2-chloroadenosine


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