title: [3H]Methoxymethyl-3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine binding to metabotropic glutamate receptor subtype 5 in rodent brain: in vitro and in vivo characterization.
abstract: The binding of [3H]methoxymethyl-3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine (methoxymethyl-MTEP), a potent and selective antagonist for metabotropic glutamate (mGlu)5 receptors, was characterized in rat brain both in vitro and in vivo. Nonspecific binding, as defined with 10 microM 2-methyl-6-(phenylethynyl)-pyridine (MPEP), was less than 10% of total binding in rat brain membranes. The binding of [3H]methoxymethyl-MTEP was of high affinity (K(d) = 20 +/- 2.7 nM), saturable (B(max) = 487 +/- 48 fmol/mg protein), and to a single site. The mGlu5 antagonists methoxymethyl-MTEP and MPEP displaced [3H]methoxymethyl-MTEP binding with IC50 values of 30 and 15 nM, respectively. In vivo administration of [3H]methoxymethyl-MTEP (50 microCi/kg i.v.) revealed 12-fold higher binding in hippocampus (an area enriched in mGlu5 receptors) relative to cerebellum (an area with few mGlu5 receptors) in rats. Similarly, administration of [3H]methoxymethyl-MTEP to mGlu5-deficient mice demonstrated binding at background levels in forebrain, whereas wild-type littermates exhibited 17-fold higher binding in forebrain relative to cerebellum. Systemic administration of unlabeled mGlu5 antagonists methoxymethyl-MTEP and MPEP to rats reduced the binding of [3H]methoxymethyl-MTEP with ID50 values of 0.8 and 2 mg/kg i.p., respectively, 1 h post-treatment. The mGlu5 agonist 2-chloro-5-hydroxyphenylglycine (CHPG) (0.3, 1, and 3 micromol) dose-dependently increased phosphoinositide (PI) hydrolysis in the hippocampus after i.c.v. administration in rats. CHPG-evoked increases in PI hydrolysis were blocked with MPEP at a dose (10 mg/kg i.p.) that markedly reduced [3H]methoxymethyl-MTEP binding in vivo. These results indicate that [3H]methoxymethyl-MTEP is a selective radioligand for labeling mGlu5 and is useful for studying the binding of mGlu5 receptors in rat brain in vitro and in vivo.
authors.affiliation: Department of Neuropharmacology, Merck Research Laboratories, San Diego, California 92121, USA. jeffery_anderson@merck.com
authors.forename: Jeffery J
authors.initials: JJ
authors.lastname: Anderson
authors.forename: Sara P
authors.initials: SP
authors.lastname: Rao
authors.forename: Blake
authors.initials: B
authors.lastname: Rowe
authors.forename: Darlene R
authors.initials: DR
authors.lastname: Giracello
authors.forename: Greg
authors.initials: G
authors.lastname: Holtz
authors.forename: Deborah F
authors.initials: DF
authors.lastname: Chapman
authors.forename: Lida
authors.initials: L
authors.lastname: Tehrani
authors.forename: Margaret J
authors.initials: MJ
authors.lastname: Bradbury
authors.forename: Nicholas D P
authors.initials: ND
authors.lastname: Cosford
authors.forename: Mark A
authors.initials: MA
authors.lastname: Varney
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000502:physiology; D001921:Brain / Q000378:metabolism*; D002531:Cerebellum / Q000378:metabolism; D018691:Excitatory Amino Acid Antagonists / Q000737:chemistry / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism; D008297:Male; D011725:Pyridines / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D064529:Receptor, Metabotropic Glutamate 5; D018094:Receptors, Metabotropic Glutamate / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism*
chemical_list: D018691:Excitatory Amino Acid Antagonists; D011725:Pyridines; D064529:Receptor, Metabotropic Glutamate 5; D018094:Receptors, Metabotropic Glutamate; D013844:Thiazoles; C469680:methoxymethyl-3-((2-methyl-1,3-thiazol-4-yl)ethynyl)pyridine; C121465:6-methyl-2-(phenylethynyl)pyridine