title: In vitro autoradiography of serotonin 5-HT(2A/2C) receptor-activated G protein: guanosine-5'-(gamma-[(35)S]thio)triphosphate binding in rat brain.
abstract: Agonist activation of G protein-coupled receptors induces an increase in the binding of guanosine 5'-(gamma-[(35)S]thio)triphosphate ([(35)S]GTPgammaS); this increase in binding has been used as a tool to investigate receptor interaction with the heterotrimer guanine nucleotide-binding regulatory protein (G protein). The present study uses agonist-stimulated [(35)S]GTPgammaS binding to characterize serotonin 5-HT(2A/2C) receptors in rat brain membrane fractions and demonstrate the anatomical localization of the receptors by in vitro autoradiography on slide-mounted sections. The stimulatory effect of the agonist [1-(2,5-dimethoxy-4-iodophenyl)]-2 aminopropane (DOI) is compared to that of serotonin (5-HT). Autoradiography revealed a similar localization of DOI- and 5-HT-stimulated binding of [(35)S]GTPgammaS in distinct areas of prefrontal and parietal cortex, consistent with previously reported 5-HT(2A) receptor distribution. Specific binding was demonstrated in the frontal and parietal cortex, medial prefrontal, and cingular and orbital-insular areas as well as in the hippocampal formation, septal areas, the nucleus accumbens, and the choroid plexus. MDL 100105, a specific 5-HT(2A) antagonist, and ketanserin, an antagonist of 5-HT(2A/2C) receptors, blocked DOI stimulation in all labeled areas, whereas 5-HT stimulation was only partially blocked (70-80%). A small but significant inhibition was observed with the specific antagonist of 5-HT(2C/2B), SB 206553. This autoradiographic technique provides a useful tool for measuring in situ changes in specific receptor-Gq protein coupling in anatomically discrete brain regions, under physiological and pathological conditions.
authors.affiliation: Department of Neuroscience, New York State Psychiatric Institute, New York, New York 10032, USA.
authors.forename: M
authors.initials: M
authors.lastname: Adlersberg
authors.forename: V
authors.initials: V
authors.lastname: Arango
authors.forename: S
authors.initials: S
authors.lastname: Hsiung
authors.forename: J J
authors.initials: JJ
authors.lastname: Mann
authors.forename: M D
authors.initials: MD
authors.lastname: Underwood
authors.forename: K
authors.initials: K
authors.lastname: Liu
authors.forename: S A
authors.initials: SA
authors.lastname: Kassir
authors.forename: D A
authors.initials: DA
authors.lastname: Ruggiero
authors.forename: H
authors.initials: H
authors.lastname: Tamir
mesh_terms: D000662:Amphetamines / Q000494:pharmacology; D000818:Animals; D001345:Autoradiography / Q000379:methods; D001923:Brain Chemistry / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D005464:Fluorobenzenes / Q000494:pharmacology; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D019204:GTP-Binding Proteins / Q000032:analysis* / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism / Q000494:pharmacology*; D006153:Guanosine Diphosphate / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted / Q000379:methods; D007211:Indoles / Q000494:pharmacology; D007650:Ketanserin / Q000494:pharmacology; D008297:Male; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D010880:Piperidines / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D011869:Radioligand Assay / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D044402:Receptor, Serotonin, 5-HT2A; D044405:Receptor, Serotonin, 5-HT2C; D011985:Receptors, Serotonin / Q000378:metabolism*; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism; D013462:Sulfur Radioisotopes
chemical_list: D000662:Amphetamines; D005464:Fluorobenzenes; D007211:Indoles; D008565:Membrane Proteins; D010880:Piperidines; D011725:Pyridines; D044402:Receptor, Serotonin, 5-HT2A; D044405:Receptor, Serotonin, 5-HT2C; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists; D013462:Sulfur Radioisotopes; D006153:Guanosine Diphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D007650:Ketanserin; C094868:SB 206553; D019204:GTP-Binding Proteins; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; C079049:volinanserin; C015952:4-iodo-2,5-dimethoxyphenylisopropylamine