title: Characterization of [(3)H]-LY354740 binding to rat mGlu2 and mGlu3 receptors expressed in CHO cells using semliki forest virus vectors.
abstract: The binding properties of [(3)H]-LY354740 were characterized on rat metabotropic glutamate receptors mGlu2 and mGlu3 expressed in Chinese hamster ovary (CHO) cells using Semliki Forest virus vectors. The saturation isotherm gave K(D) values of 20+/-5 and 53+/-8 nM and B(max) values of 474+/-161 and 667+/-89 fmol/mg protein for mGlu2 and mGlu3 receptors, respectively. NMDA, CaCl(2), DHPG and kainate were inactive up to 1 mM, whereas LY341495, DCG IV and ibotenate inhibited [(3)H]-LY354740 binding with similar potencies on both receptors. L-CCG I, L-AP4, L-AP5, LY354740 and 1S,3R-ACPD were 2- to 4-fold more potent inhibitors of [(3)H]-LY354740 binding to mGlu2 than mGlu3 receptors. However, MPPG and L-AP3 had a 6-fold and DTT a 28-fold preference for mGlu2 over mGlu3. ZnCl(2), at 10 mM, inhibited more than 70% of [(3)H]-LY354740 binding to mGlu2 receptors. At the same concentration it did not affect significantly [(3)H]-LY354740 binding to mGlu3 receptors. On the contrary, glutamate, quisqualate, EGLU and NAAG showed a 3-, 5-, 7- and 12-fold preference for mGlu3 over mGlu2. Finally, GTPgammaS, which partially inhibited the binding on mGlu2 receptors, was inactive to inhibit [(3)H]-LY354740 binding on mGlu3 receptors.
authors.affiliation: Pharmaceutical Division, Preclinical CNS Research, F. Hoffmann-La Roche Ltd., CH-4070, Basel, Switzerland.
authors.forename: C
authors.initials: C
authors.lastname: Schweitzer
authors.forename: C
authors.initials: C
authors.lastname: Kratzeisen
authors.forename: G
authors.initials: G
authors.lastname: Adam
authors.forename: K
authors.initials: K
authors.lastname: Lundstrom
authors.forename: P
authors.initials: P
authors.lastname: Malherbe
authors.forename: S
authors.initials: S
authors.lastname: Ohresser
authors.forename: H
authors.initials: H
authors.lastname: Stadler
authors.forename: J
authors.initials: J
authors.lastname: Wichmann
authors.forename: T
authors.initials: T
authors.lastname: Woltering
authors.forename: V
authors.initials: V
authors.lastname: Mutel
mesh_terms: D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D001643:Bridged Bicyclo Compounds / Q000378:metabolism*; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D002712:Chlorides / Q000494:pharmacology; D006224:Cricetinae; D004274:DNA, Recombinant / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D015870:Gene Expression; D005822:Genetic Vectors; D018698:Glutamic Acid / Q000494:pharmacology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D007700:Kinetics; D051381:Rats; D018094:Receptors, Metabotropic Glutamate / Q000235:genetics / Q000378:metabolism*; D012672:Semliki forest virus / Q000235:genetics; D014316:Tritium; D014966:Xanthenes / Q000494:pharmacology; D017967:Zinc Compounds / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D001643:Bridged Bicyclo Compounds; D002712:Chlorides; D004274:DNA, Recombinant; D018690:Excitatory Amino Acid Agonists; D018691:Excitatory Amino Acid Antagonists; C114624:LY 341495; D018094:Receptors, Metabotropic Glutamate; D014966:Xanthenes; D017967:Zinc Compounds; C108822:metabotropic glutamate receptor 2; C102718:metabotropic glutamate receptor 3; D014316:Tritium; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D018698:Glutamic Acid; C016837:zinc chloride; C104753:eglumetad