In vivo evidence for dopamine-mediated internalization of D2-receptors after amphetamine: differential findings with [3H]raclopride versus [3H]spiperone

Abstract

Competition with endogenous dopamine (DA) is usually invoked to explain changes in [(11)C]raclopride binding observed after amphetamine administration in animals and humans. This account has recently been questioned because a number of inconsistencies have been reported that contradict it. In the present study, we investigated whether the decrease in [(3)H]raclopride binding observed in the rat striatum after an amphetamine challenge reflects true competition with endogenous DA or agonist-mediated internalization of D(2)-receptors. We found that the amphetamine-induced decrease in [(3)H]raclopride binding is caused by a decrease in D(2)-receptor density (B(max)) with no change in affinity (K(d)). In contrast, in the same tissue, neither the B(max) nor the K(d) were affected when measured with [(3)H]spiperone. Challenge with amphetamine not only decreased the number of D(2)-receptors but also eliminated the proportion (22%) of receptors usually in the high-affinity state. The addition of Gpp(NH)p had no effect on B(max), suggesting that these receptors were not just noncompetitively bound with dopamine at the cell-surface. Subcellular fractionation studies showed that amphetamine treatment led to a decrease in radioligand binding in the cell-surface fraction for both [(3)H]raclopride and [(3)H]spiperone; however, in the case of [(3)H]spiperone, this was accompanied by a compensatory increase in binding in the intracellular compartment, whereas no increase was seen with [(3)H]raclopride. These data suggest that amphetamine releases dopamine, which binds to the high-affinity state of the D(2)-receptor, leading to its sequestration in some intracellular compartment; in this compartment, sequestered receptors are inaccessible to [(3)H]raclopride binding but can still be bound by [(3)H]spiperone.