Unraveling the identity of benzodiazepine binding sites in rat hipppocampus and olfactory bulb.
The goals of the work reported here were (i) to identify distinct GABA(A)/benzodiazepine receptors in the rat hippocampus and olfactory bulb using receptor binding assays, and (ii) to determine the affinities and selectivities of benzodiazepine receptor ligands from structurally diverse chemical families at each site identified. These studies were aided by the use of software AFFINITY ANALYSIS SYSTEM, developed in our laboratory for analysis of receptor binding data that allows the determination of receptor heterogeneity using non-selective radioligands. Saturation binding assays using [3H]RO15-4513 (ethyl 8-azido-6-dihydro-5-methyl-6-oxo-4H-imidazo[1, 5-a]-[1,4]benzodiazepine-3-carboxylate) revealed two binding sites in each of these two tissues. The higher affinity site corresponds to alpha(5) subunit-containing GABA(A) receptor and the lower affinity site to a combination of alpha(1), alpha(2), and alpha(3) subunit-containing receptors. These results should be useful in the challenging task of identifying the various functional GABA(A) receptors in the central nervous system, and in providing a link between receptor affinities and in vivo activities of the GABA(A)/benzodiazepine receptor ligands studied.
Lameh
J
J
Molecular Research Institute, 2495 Old Middlefield Way, Mountain View, CA 94034-2316, USA. jlameh@purisima.molres.org
Wang
P
P
Elgart
D
D
Meredith
D
D
Shafer
S L
SL
Loew
G H
GH
D000818:Animals; D001569:Benzodiazepines / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D002243:Carbolines / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism*; D007093:Imidazoles / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D009830:Olfactory Bulb / Q000378:metabolism*; D011720:Pyrazoles / Q000378:metabolism; D011725:Pyridines / Q000378:metabolism; D011804:Quinolines / Q000378:metabolism; D011810:Quinoxalines / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D011963:Receptors, GABA-A / Q000378:metabolism*; D013329:Structure-Activity Relationship
D002243:Carbolines; D007093:Imidazoles; D008024:Ligands; D011720:Pyrazoles; D011725:Pyridines; D011804:Quinolines; D011810:Quinoxalines; D011963:Receptors, GABA-A; D001569:Benzodiazepines