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Pharmacological properties of recombinant human N-methyl-D-aspartate receptors comprising NR1a/NR2A and NR1a/NR2B subunit assemblies expressed in permanently transfected mouse fibroblast cells.
Mol Pharmacol. 1995 Nov; 48(5):841-8.MP

Abstract

The pharmacological properties of two recombinant human N-methyl-D-aspartate (NMDA) receptor subtypes, comprising either NR1a/NR2A or NR1a/NR2B subunits permanently transfected into mouse L(tk-) cells, have been compared using whole-cell voltage-clamp electrophysiology. Glutamate was a full agonist at both receptors, having a modestly but statistically significant (p < 0.002) higher affinity for the NR2B- than the NR2A-containing receptor (microscopic Kd [mKd] = 0.76 and 0.43 microM, respectively). In comparison to glutamate, NMDA, quinolinic acid, and cis-2,3-piperidinedicarboxylic acid were partial agonists at both receptor subtypes. Maximal amplitude currents resulted when glutamate-site agonists were combined with either glycine or D-serine; both of these amino acids were, therefore, defined as full agonists at the glycine site. Glycine had an approximately 10-fold higher affinity (p < 0.0001) for NR2B- than for NR2A-containing receptors (mKd = 0.057 and 0.53 microM, respectively). D-Cycloserine, (+)-(3R)-3-amino-1-hydroxypyrrolidin-2-one, (+)-cis-(4R)-methyl-(3R)-amino-1-hydroxypyrrolidin-2-one, and 1-aminocyclobutanecarboxylic acid also had higher affinities for the NR2B-containing receptor but were partial agonists, at both receptor subtypes, unlike glycine. Agonist-evoked whole-cell currents were antagonized by D-(-)-2-amino-5-phosphonopentanoic acid, cis-4-(phosphonomethyl)piperidine-2-carboxylic acid, and 3-((R)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid, all of which had slightly, but statistically significant, higher affinities (2.2-, 2.8-, and 5.5-fold, respectively) for the NR2A-containing receptor. Responses were also antagonized by the glycine-site antagonists 7-chlorokynurenic acid, 7-chloro-4-hydroxy-3-(3-phenoxy)phenylquinolin-2-(1H)-one, and (+/=)-4-(trans)-2-carboxy-5,7-dichloro-4-phenylaminocarbonylamino- 1,2,3,4- tetrahydroquinoline. The atypical NMDA antagonist ifenprodil showed the largest separation in functional affinity (IC50 values, 0.6 and 175 microM at NR2B- and NR2A-containing receptors, respectively). These experiments demonstrate the usefulness of permanently transfected L(tk-) cells for electrophysiological studies of recombinant NMDA receptor function and provide the first detailed functional pharmacological analysis of human NMDA receptor subtypes.

Authors+Show Affiliations

Merck, Sharp & Dohme Research Laboratories, Neuroscience Research Centre, Harlow, Essex, UK.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

7476914

Citation

Priestley, T, et al. "Pharmacological Properties of Recombinant Human N-methyl-D-aspartate Receptors Comprising NR1a/NR2A and NR1a/NR2B Subunit Assemblies Expressed in Permanently Transfected Mouse Fibroblast Cells." Molecular Pharmacology, vol. 48, no. 5, 1995, pp. 841-8.
Priestley T, Laughton P, Myers J, et al. Pharmacological properties of recombinant human N-methyl-D-aspartate receptors comprising NR1a/NR2A and NR1a/NR2B subunit assemblies expressed in permanently transfected mouse fibroblast cells. Mol Pharmacol. 1995;48(5):841-8.
Priestley, T., Laughton, P., Myers, J., Le Bourdellés, B., Kerby, J., & Whiting, P. J. (1995). Pharmacological properties of recombinant human N-methyl-D-aspartate receptors comprising NR1a/NR2A and NR1a/NR2B subunit assemblies expressed in permanently transfected mouse fibroblast cells. Molecular Pharmacology, 48(5), 841-8.
Priestley T, et al. Pharmacological Properties of Recombinant Human N-methyl-D-aspartate Receptors Comprising NR1a/NR2A and NR1a/NR2B Subunit Assemblies Expressed in Permanently Transfected Mouse Fibroblast Cells. Mol Pharmacol. 1995;48(5):841-8. PubMed PMID: 7476914.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Pharmacological properties of recombinant human N-methyl-D-aspartate receptors comprising NR1a/NR2A and NR1a/NR2B subunit assemblies expressed in permanently transfected mouse fibroblast cells. AU - Priestley,T, AU - Laughton,P, AU - Myers,J, AU - Le Bourdellés,B, AU - Kerby,J, AU - Whiting,P J, PY - 1995/11/1/pubmed PY - 1995/11/1/medline PY - 1995/11/1/entrez SP - 841 EP - 8 JF - Molecular pharmacology JO - Mol Pharmacol VL - 48 IS - 5 N2 - The pharmacological properties of two recombinant human N-methyl-D-aspartate (NMDA) receptor subtypes, comprising either NR1a/NR2A or NR1a/NR2B subunits permanently transfected into mouse L(tk-) cells, have been compared using whole-cell voltage-clamp electrophysiology. Glutamate was a full agonist at both receptors, having a modestly but statistically significant (p < 0.002) higher affinity for the NR2B- than the NR2A-containing receptor (microscopic Kd [mKd] = 0.76 and 0.43 microM, respectively). In comparison to glutamate, NMDA, quinolinic acid, and cis-2,3-piperidinedicarboxylic acid were partial agonists at both receptor subtypes. Maximal amplitude currents resulted when glutamate-site agonists were combined with either glycine or D-serine; both of these amino acids were, therefore, defined as full agonists at the glycine site. Glycine had an approximately 10-fold higher affinity (p < 0.0001) for NR2B- than for NR2A-containing receptors (mKd = 0.057 and 0.53 microM, respectively). D-Cycloserine, (+)-(3R)-3-amino-1-hydroxypyrrolidin-2-one, (+)-cis-(4R)-methyl-(3R)-amino-1-hydroxypyrrolidin-2-one, and 1-aminocyclobutanecarboxylic acid also had higher affinities for the NR2B-containing receptor but were partial agonists, at both receptor subtypes, unlike glycine. Agonist-evoked whole-cell currents were antagonized by D-(-)-2-amino-5-phosphonopentanoic acid, cis-4-(phosphonomethyl)piperidine-2-carboxylic acid, and 3-((R)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid, all of which had slightly, but statistically significant, higher affinities (2.2-, 2.8-, and 5.5-fold, respectively) for the NR2A-containing receptor. Responses were also antagonized by the glycine-site antagonists 7-chlorokynurenic acid, 7-chloro-4-hydroxy-3-(3-phenoxy)phenylquinolin-2-(1H)-one, and (+/=)-4-(trans)-2-carboxy-5,7-dichloro-4-phenylaminocarbonylamino- 1,2,3,4- tetrahydroquinoline. The atypical NMDA antagonist ifenprodil showed the largest separation in functional affinity (IC50 values, 0.6 and 175 microM at NR2B- and NR2A-containing receptors, respectively). These experiments demonstrate the usefulness of permanently transfected L(tk-) cells for electrophysiological studies of recombinant NMDA receptor function and provide the first detailed functional pharmacological analysis of human NMDA receptor subtypes. SN - 0026-895X UR - https://www.unboundmedicine.com/medline/citation/7476914/Pharmacological_properties_of_recombinant_human_N_methyl_D_aspartate_receptors_comprising_NR1a/NR2A_and_NR1a/NR2B_subunit_assemblies_expressed_in_permanently_transfected_mouse_fibroblast_cells_ L2 - http://molpharm.aspetjournals.org/cgi/pmidlookup?view=long&amp;pmid=7476914 DB - PRIME DP - Unbound Medicine ER -