GTP differentially affects antagonist radioligand binding to adenosine A(1) and A(2A) receptors in human brain

The effect of guanosine triphosphate (GTP) on the interaction of antagonist s with human adenosine A(1) and A(2A) receptors was studied using whole-hem isphere sections from human brain and membranes from Chinese hamster ovary (CHO) cells expressing human A(1) and A(2A) receptors. Adenosine A(1) recep tors, studied using [H-3]1,3-dipropyl-8-cyclopentylxanthine ([H-3]DPCPX) as radioligand, showed the expected regional distribution in human brain. Add ition of 500 mu M GTP significantly increased (23-55%) [H-3] DPCPX binding in all regions measured. In CHO cells transfected with human adenosine A(1) receptor cDNA, the number of receptors, B-max, increased from 401 (359-442 ) to 667 (592-743) fmol/mg protein upon addition of GTP. [H-3]5-Amino-7-(2- phenylethyl)-2-(2-furyl)pyrazolo- [4,3-e]-1,2,4-triazolo-[1,5-c]-pyrimidin e ([H-3]SCH 58261), a selective adenosine A(2A) receptor ligand, showed sat urable binding to membranes from CHO cells transfected with adenosine A(2A) receptor cDNA and was localized to striatum and globus pallidus in human b rain sections. Addition of GTP did not significantly change [H-3]SCH 58261 binding to brain sections or CHO cell membranes. These results indicate tha t human A(1) and A(2A) receptors are not substantially different from those of the rat as regards regulation by GTP and interactions with endogenous a denosine in binding experiments. However, the relative abundance of the rec eptors differs between species, and this may be related to the differences observed in the potency of the endogenous agonist. (C) 2000 Elsevier Scienc e Ltd. All rights reserved.