MOLECULAR-CLONING, FUNCTIONAL EXPRESSION AND PHARMACOLOGICAL CHARACTERIZATION OF THE HUMAN METABOTROPIC GLUTAMATE-RECEPTOR TYPE-2

被引:64
作者
FLOR, PJ [1 ]
LINDAUER, K [1 ]
PUTTNER, I [1 ]
RUEGG, D [1 ]
LUKIC, S [1 ]
KNOPFEL, T [1 ]
KUHN, R [1 ]
机构
[1] CIBA GEIGY AG,CNS RES,DEPT MOLEC & CELLULAR BIOL,CH-4002 BASEL,SWITZERLAND
关键词
CAMP; PHENYLGLYCINE; CHINESE HAMSTER OVARY CELLS; IMMUNOCYTOCHEMISTRY; RAT;
D O I
10.1111/j.1460-9568.1995.tb00666.x
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
A cDNA encoding the human metabotropic glutamate receptor type 2 (hmGluR2) was isolated from human brain cDNA libraries by cross-hybridization with rat mGluR2 probes. The deduced amino acid sequence of the human mGluR2 receptor consists of 872 residues and shows a sequence identity of 97% to the amino acid sequence of rat mGluR2. Northern blot analyses showed that hmGluR2 is widely expressed in different regions of the adult brain as well as in fetal human brain. Genomic Southern blotting localized the mGluR2 gene to human chromosome 3. Chinese hamster ovary (CHO) cells stably transfected with the cloned hmGluR2 cDNA exhibit agonist induced depression of forskolin-stimulated cAMP accumulation. A direct comparison of CHO cells stably expressing human and rat mGluR2 with five agonists revealed the same rank order of potency [(2S,3S,4S)-(alpha(carboxycyclopropyl)-glycine >> (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid = L-glutamate >> quisqualate = L-2-amino-4-phosphonobutyric acid] and similar EC(50) values for both homologous receptors. (R,S)-alpha-methyl-4-carboxyphenylglycine, a reported antagonist at some mGluR subtypes, reduced the depression of forskolin-induced cAMP accumulation by (1S,3R)-ACPD in both human and rat mGluR2.
引用
收藏
页码:622 / 629
页数:8
相关论文
共 42 条
  • [1] Abe T., Sugihara H., Nawa H., Shigemoto R., Mizuno N., Nakanishi S., Molecular characterization of a novel metabotropic glutamate receptor mGluR5 coupled to inositol phosphate/Ca<sup>2+</sup> signal transduction, J. Biol. Chem., 267, pp. 13361-13368, (1992)
  • [2] Aramori I., Nakanishi S., Signal transduction and pharmacological characteristics of a metabotropic glutamate receptor, mGluR1, in transfected CHO cells, Neuron, 8, pp. 757-765, (1992)
  • [3] Asselbergs F.A.M., Grand P., A two‐plasmid system for transient expression of cDNAs in primate cells, Anal. Biochem., 209, pp. 327-331, (1993)
  • [4] Bashir Z.I., Bortolotto Z.A., Davies C.H., Berretta N., Irving A.J., Seal A.J., Henley J.M., Jane D.E., Watkins J.C., Collingridge G.L., Induction of LTP in the hippocampus needs synaptic activation of glutamate metabotropic receptors, Nature, 363, pp. 347-350, (1993)
  • [5] Bockaert J., Pin J., Fagni L., Metabotropic glutamate receptors, an original family of G protein‐coupled receptors, Fundam. Clin. Pharmacol., 7, pp. 473-485, (1993)
  • [6] Calabresi P., Mercuri N., Bernardi G., Activation of quisqualate metabotropic receptors reduced glutamate and GABA‐mediated synaptic potentials in the striatum, Neurosci. Lett., 139, pp. 41-44, (1992)
  • [7] Charpak S., Gahwiler B.H., Do K.Q., Knopfel T., Potassium conductances in hippocampal neurons blocked by excitatory amino‐acid transmitters, Nature, 347, pp. 765-767, (1990)
  • [8] Chomczynski P., Sacci N., Single‐step method of RNA isolation by acid guanidinium thiocyanate‐phenol‐chloroform extraction, Anal. Biochem., 162, pp. 156-159, (1987)
  • [9] Desai M.A., Smith T.S., Conn P.J., Multiple metabotropic glutamate receptors regulate hippocampal function, Synapse, 12, pp. 206-213, (1992)
  • [10] Devereux J., Haeberl P., Smithies O., A comprehensive set of sequence analysis programs for the VAX, Nucleic Acids Res., 12, pp. 387-395, (1984)