title: Demonstration of competition between endogenous dopamine and [11C]raclopride binding in in vitro brain slices using a dynamic autoradiography technique.
abstract: To elucidate the mechanism of in vivo binding competition between radioligand and endogenously released transmitter, we examined the influence of depolarization-induced dopamine (DA) release on [11C]raclopride-specific binding to D2 receptors in slices of living brain tissues using dynamic positron autoradiography. Rat brain slices were incubated in a chamber with [11C]raclopride in oxygenated medium at 34 degrees C for 150 min. Two-dimensional images of radioactivity in the slices were recorded on a storage phosphor screen and dynamic changes were measured. When the brain slices were exposed to the depolarization agents (25 mM K+, 50 mM K+, and 20 microM veratridine), the percentage inhibition of striatal [11C]raclopride-specific binding was 22 +/- 4%, 44 +/- 8% and 54 +/- 7% of the control, respectively. The percentage inhibition of [11C]raclopride-specific binding during each depolarization treatment agreed proportionally with the amount of DA released into the medium. However, preexposure of brain slices to the same depolarization treatment (50 mM K+) did not affect the [11C]raclopride-specific binding, suggesting that the reduction in receptor density and/or affinity was not involved in the decrease of [11C]raclopride-specific binding. [11C]Raclopride-specific binding decreased dose-dependently in the presence of exogenously added DA (range 0.005-3mM). The synaptic DA concentration during each depolarization treatment estimated using three different methods. These results suggest that the decrease of [11C]raclopride-specific binding to striatal slices following evoked DA release is due to competition between endogenous DA and raclopride. It is unlikely that changes in D(2) receptor density or in affinity of the receptors for raclopride are involved. These results provide supportive evidence for in vivo binding competition between radioligand and endogenous neurotransmitter.
authors.affiliation: Positron Medical Center, Tokyo Metropolitan Institute of Gerontology, Tokyo 173-0022, Japan. sasaki@pet.tmig.or.jp
authors.forename: Toru
authors.initials: T
authors.lastname: Sasaki
authors.forename: Kiichi
authors.initials: K
authors.lastname: Ishiwata
authors.forename: Tetsuhito
authors.initials: T
authors.lastname: Murata
authors.forename: Michio
authors.initials: M
authors.lastname: Senda
mesh_terms: D000818:Animals; D001345:Autoradiography; D001667:Binding, Competitive / Q000187:drug effects* / Q000502:physiology; D001921:Brain / Q000000981:diagnostic imaging / Q000187:drug effects* / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D018492:Dopamine Antagonists / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D008297:Male; D011188:Potassium / Q000494:pharmacology; D017729:Presynaptic Terminals / Q000000981:diagnostic imaging / Q000187:drug effects* / Q000378:metabolism; D020891:Raclopride / Q000378:metabolism*; D011869:Radioligand Assay; D011877:Radionuclide Imaging; D051381:Rats; D017208:Rats, Wistar; D009435:Synaptic Transmission / Q000187:drug effects* / Q000502:physiology; D014701:Veratridine / Q000494:pharmacology
chemical_list: D018492:Dopamine Antagonists; D020891:Raclopride; D014701:Veratridine; D011188:Potassium; D004298:Dopamine