Sodium-adenosine cotransport in brush-border membranes from rabbit ileum

Abstract

To determine the mechanism(s) of transcellular adenosine transport in epithelial tissues that possess an adenosine receptor response, we studied [3H]adenosine uptake using vesicles prepared from isolated brush-border and basolateral membranes of the rabbit ileum. In the presence of the adenosine deaminase inhibitor deoxycoformycin uptake of [3H]adenosine into brush-border membrane vesicles is stimulated fivefold by an inwardly directed Na gradient. Na-dependent [3H]adenosine uptake is enhanced and concentrative under conditions that increase inside negativity of vesicles, thus providing evidence for an electrogenic carrier. Na-dependent adenosine uptake is a saturable function of adenosine concentration with a Michaelis-Menten constant of 17.3 +/- 7.1 microM and maximum transport rate of 216.9 +/- 20.2 pmol.min-1.mg protein-1. Both uridine and inosine inhibit [3H]adenosine uptake, suggesting that the Na-dependent transporter has broad substrate specificity for both purine and pyrimidine ribonucleosides. Na-dependent adenosine uptake is inhibited by dipyridamole but is insensitive to 6-(4-nitrobenzyl)thio-9-beta-D-ribofuranosylpurine. We conclude that adenosine is transported across ileal brush-border membranes by a Na-ribonucleoside cotransport system. In contrast, adenosine uptake in basolateral membranes is not stimulated by a Na gradient. These studies show asymmetry in the distribution of transport systems for adenosine in polarized intestinal epithelia.