THE KINETICS OF [H-3] SCH-23390 DISSOCIATION FROM RAT STRIATAL DOPAMINE D-1 RECEPTORS - EFFECT OF DOPAMINE

Citation
Cb. Farrell et Km. Oboyle, THE KINETICS OF [H-3] SCH-23390 DISSOCIATION FROM RAT STRIATAL DOPAMINE D-1 RECEPTORS - EFFECT OF DOPAMINE, European journal of pharmacology. Molecular pharmacology section, 268(1), 1994, pp. 79-88
Citations number
30
Categorie Soggetti
Pharmacology & Pharmacy
ISSN journal
09224106
Volume
268
Issue
1
Year of publication
1994
Pages
79 - 88
Database
ISI
SICI code
0922-4106(1994)268:1<79:TKO[SD>2.0.ZU;2-5
Abstract
The present study investigated possible allosteric interactions betwee n dopamine and [H-3]SCH 23390 o-3-methyl-5-phenyl-1H-3-benzazepin-7-ol )-labelled dopamine D-1 receptors in rat striatum. As previously descr ibed, dopamine prevented [H-3]SCH 23390 binding in a mixed competitive /non-competitive manner, causing both a loss of ligand affinity and a decrease in B-max. The effect of dopamine was largely reversed followi ng pretreatment of the membranes with 100 mu M Gpp(NH)p (5'-guanylylim idodiphosphate) and was significantly enhanced by omission of Na+ from the incubation buffer. In dissociation kinetic studies, two methods o f initiating ligand dissociation were used: dilution into 100-fold vol ume excess of buffer or addition of a molar excess of drug. Both metho ds yielded similar rates of [H-3]SCH 23390 dissociation. Inclusion of dopamine in the volume excess of buffer did not alter the k(-1) for [H -3]SCH 23390 dissociation. However, when 100 mu M dopamine was used in stead of 1 mu M piflutixol to initiate dissociation, a significant slo wing of the rate of dissociation of [H-3]SCH 23390 occurred. This effe ct of dopamine on k(-1) was Na+-dependent since in the absence of Nathe dopamine-induced rate of dissociation was only slightly slower tha n control values. Under neither condition did dopamine accelerate the rate of ligand dissociation, indicating that dopamine does not interac t allosterically with [H-3]SCH 23390 binding sites. These data, theref ore, preclude an allosteric mechanism to explain the dopamine-induced decrease in dopamine D-1 receptor density and provide direct evidence that dopamine masks ligand binding by binding to a high affinity site which can be modulated by Gpp(NH)p and Na+.