Effector coupling of stably transfected human A3 adenosine receptors in CHO cells.
CHO cells stably transfected with adenosine receptors are widely utilized models for binding and functional studies. The effector coupling of human A3 adenosine receptors expressed in such a cellular model was characterized. Inhibition of adenylyl cyclase via a pertussis toxin-sensitive G protein was confirmed and exhibited a pharmacological profile in accordance with agonist binding data. The agonist potency was dependent on the assay system utilized to measure cyclase inhibition. Agonists were more potent in a cell-based assay than in experiments where cyclase inhibition was measured in a membrane preparation suggesting that receptor-effector coupling might be more efficient in intact cells. In addition to the modulation of cyclase activity, stimulation of A3 receptors elicited a Ca2+ response in CHO cells with agonist potencies corresponding to the values for the whole cell cAMP assay. The Ca2+ signal was completely eliminated by pertussis toxin treatment suggesting that it is mediated via betagamma release from a heterotrimeric G protein of the Gi/o family. These results show that cAMP and Ca2+ signaling characteristics of the A3 adenosine receptor are comparable to the ones found for the A1 subtype.
Englert
Martin
M
Institut für Pharmakologie und Toxikologie, Universität Würzburg, Versbacher Str. 9, Würzburg, Germany.
Quitterer
Ursula
U
Klotz
Karl Norbert
KN
D000818:Animals; D016466:CHO Cells; D020013:Calcium Signaling / Q000502:physiology*; D006224:Cricetinae; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D043684:Receptor, Adenosine A3; D018047:Receptors, Purinergic P1 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D043684:Receptor, Adenosine A3; D018047:Receptors, Purinergic P1; D019204:GTP-Binding Proteins