title: Regional differences in cannabinoid receptor/G-protein coupling in rat brain.
abstract: Cannabinoid receptor activation of G-proteins can be measured by WIN 55212-2-stimulated [35S]GTPgammaS binding. Receptor/transducer amplification factors, interpreted as the number of G-proteins activated per occupied receptor, are the ratio of the apparent B(max) of net agonist-stimulated [35S]GTPgammaS binding to the B(max) of receptor binding. The present study examined whether amplification factors for cannabinoid receptors differ among various rat brain regions. In autoradiographic studies with [3H]WIN 55212-2 and WIN 55212-2-stimulated [35S]GTPgammaS binding, some regions displayed different relative levels of agonist-stimulated [35S]GTPgammaS binding than receptor binding. To quantify amplification factors, membranes from different brain regions were assayed by saturation binding analysis of net WIN 55212-2-stimulated [35S]GTPgammaS, [3H]SR141716A (antagonist) and [3H]WIN 55212-2 (agonist) binding. For [3H]SR141716A binding, amplification factors varied significantly from 2.0 (frontal cortex) to 7.5 (hypothalamus). For [3H]WIN 55212-2 binding, amplification factors ranged from 2.4 (hippocampus) to 5.5 (thalamus). Comparison of receptor binding and G-protein activation at subsaturating concentrations of WIN 55212-2 indicated that amplification factors may vary with receptor occupancy in some regions like cerebellum. Ratios between high-affinity [3H]WIN 55212-2 B(max) and [3H]SR141716A B(max) also differed significantly among brain regions. These results demonstrate that G-protein coupling by cannabinoid receptors differs among brain regions, and therefore depends on the cellular environment.
authors.affiliation: Department of Physiology and Pharmacology, and Center for Investigative Neuroscience, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27157, USA.
authors.forename: C S
authors.initials: CS
authors.lastname: Breivogel
authors.forename: L J
authors.initials: LJ
authors.lastname: Sim
authors.forename: S R
authors.initials: SR
authors.lastname: Childers
mesh_terms: D000818:Animals; D001345:Autoradiography; D048588:Benzoxazines; D001921:Brain / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D008297:Male; D009025:Morpholines / Q000378:metabolism; D009281:Naphthalenes / Q000378:metabolism; D010880:Piperidines / Q000378:metabolism; D011720:Pyrazoles / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D043882:Receptors, Cannabinoid; D011955:Receptors, Drug / Q000378:metabolism*; D000077285:Rimonabant
chemical_list: D048588:Benzoxazines; D009025:Morpholines; D009281:Naphthalenes; D010880:Piperidines; D011720:Pyrazoles; D043882:Receptors, Cannabinoid; D011955:Receptors, Drug; D016244:Guanosine 5'-O-(3-Thiotriphosphate); C070417:(3R)-((2,3-dihydro-5-methyl-3-((4-morpholinyl)methyl)pyrrolo-(1,2,3-de)-1,4-benzoxazin-6-yl)(1-naphthalenyl))methanone; D019204:GTP-Binding Proteins; D000077285:Rimonabant