Characterization of dopamine binding sites in standard preparations of brain synaptic membranes

Autor: Melvin Blecher, George Bers
Rok vydání: 1977
Předmět:
Zdroj: Biochemical pharmacology. 26(21)
ISSN: 0006-2952
Popis: Brain synaptic membranes, prepared according to DeRobertis and to Whittaker, were compared morphologically and biochemically. Electron microscopy revealed that DeRobertis preparations were heavily contaminated with synaptosomes, mitochondria, storage vesicles, and a variety of extraneous membrane structures; Whittaker preparations appeared to consist of synaptic membranes, with a relatively small number of presynaptic storage vesicles. Substantial dopamine β-hydroxylase, monoamine oxidase, and catecholamine- O -methyl transferase activities were present in DeRobertis preparations; Whittaker membrane preparations contained low-to-undetectable activities of these enzymes. Although dopamine bound to membranes derived from both brain cortex and corpus striatum, only membranes from the latter area contained dopamine-sensitive adenylate cyclase. Binding of dopamine to Whittaker striatal synaptic membrane preparations was relatively rapid, saturable, partially reversible, and did not chemically alter the ligand. A variety of dopamine acceptors appeared to be present in this preparation since the ed 50 (from Klotz plots) of dopamine binding was 25 times that for activation of adenylate cyclase; Scatchard plots revealed both high and low affinity binding sites for dopamine; and, binding and adenylate cyclase activation studies with dopamine, carried out in the presence of fluphenazine, cocaine, pargyline, and reserpine, reveal that even the more-pure Whittaker preparations of striatal synaptic membranes contain at least three major dopamine-binding components: postsynaptic (and perhaps presynaptic as well) membrane receptors linked to adenylate cyclase, a presynaptic synaptosomal reuptake receptor, and storage vesicle sequestration.
Databáze: OpenAIRE