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tk;1Adenosine plays a role in the control of water and electrolyte reabsorption in the distal tubule. As the distal convoluted tubule is important in the regulation of renal Mg 2+  balance, we determined the effects of adenosine on cellular Mg 2+  uptake in this segment. The effect of adenosine was studied on immortalized mouse distal convoluted tubule (MDCT) cells, a model of the intact distal convoluted tubule. The rate of Mg 2+  uptake was measured with fluorescence techniques using mag-fura 2. To assess Mg 2+  uptake, MDCT cells were first Mg 2+  depleted to 0.22 ± 0.01 mM by being cultured in Mg 2+ -free media for 16 h and then placed in 1.5 mM MgCl 2 ; next, changes in intracellular Mg 2+  concentration ([Mg 2+ ] i ) were determined. [Mg 2+ ] i  returned to basal levels, 0.53 ± 0.02 mM, with a mean refill rate, d([Mg 2+ ] i )/d t, of 137 ± 16 nM/s. Adenosine stimulates basal Mg 2+  uptake by 41 ± 10%. The selective A 1  purinoceptor agonist N  6 -cyclopentyladenosine (CPA) increased intracellular Ca 2+  and decreased parathyroid hormone (PTH)-stimulated cAMP formation and PTH-mediated Mg 2+ uptake. On the other hand, the selective A 2  receptor agonist 2-[ p-(2-carbonyl-ethyl)-phenylethylamino]-5′- N-ethylcarboxamidoadenosine (CGS) stimulated Mg 2+  entry in a concentration-dependent fashion. CGS increased cAMP formation and the protein kinase A inhibitor RpcAMPS inhibited CGS-stimulated Mg 2+  uptake. Selective inhibition of phospholipase C, protein kinase C, or mitogen-activated protein kinase enzyme cascades with U-73122, Ro-31-8220, and PD-98059, respectively, diminished A 2  agonist-mediated Mg 2+  entry. Aldosterone potentiated CGS-mediated Mg 2+  entry, and elevation of extracellular Ca 2+  diminished CGS-responsive cAMP formation and Mg 2+  uptake. Accordingly, MDCT cells possess both A 1  and A 2  purinoceptor subtypes with intracellular signaling typical of these respective receptors. We conclude that adenosine has dual effects on Mg 2+  uptake in MDCT cells through separate A 1  and A 2 purinoceptor pathways.

Adenosine modulates Mg2+ uptake in distal convoluted tubule cells via A1 and A2purinoceptors

Adenosine modulates Mg²⁺ uptake in distal convoluted tubule cells via A₁ and A₂purinoceptors