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Membrane Transport Research Group, Departments of 1 Physiology and 3 Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 2H7; and 2 School of Biochemistry and Molecular Biology, University of Leeds, Leeds, LS2 9JT, United Kingdom
The regulatory actions of adenosine on ion channel function are mediated by four distinct membrane receptors. The concentration of adenosine in the vicinity of these receptors is controlled, in part, by inwardly directed nucleoside transport. The purpose of this study was to characterize the effects of adenosine on ion channels in A549 cells and the role of nucleoside transporters in this regulation. Ion replacement and pharmacological studies showed that adenosine and an inhibitor of human equilibrative nucleoside transporter (hENT)-1, nitrobenzylthioinosine, activated K+ channels, most likely Ca2+-dependent intermediate-conductance K+ (IK) channels. A1 but not A2 receptor antagonists blocked the effects of adenosine. RT-PCR studies showed that A549 cells expressed mRNA for IK-1 channels as well as A1, A2A, and A2B but not A3 receptors. Similarly, mRNA for equilibrative (hENT1 and hENT2) but not concentrative (hCNT1, hCNT2, and hCNT3) nucleoside transporters was detected, a result confirmed in functional uptake studies. These studies showed that adenosine controls the function of K+ channels in A549 cells and that hENTs play a crucial role in this process.
A549 cells; nitrobenzylthioinosine; human concentrative nucleoside transporter; human equilibrative nucleoside transporter; calcium-dependent intermediate-conductance potassium channels
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