Document Detail

Generation and characterisation of stable cell lines expressing recombinant human N-methyl-D-aspartate receptor subtypes.
MedLine Citation:
PMID:  8632144     Owner:  NLM     Status:  MEDLINE    
Transfection of mouse L(tk-) cells with human N-methyl-D-aspartate (NMDA) receptor subunit cDNAs under the control of a dexamethasone-inducible promoter has been used to generate two stable cell lines expressing NR1a/NR2A receptors and a stable cell line expressing NR1a/NR2B receptors. The cell lines have been characterised by northern and western blot analyses, and the pharmacology of the recombinant receptors determined by radioligand binding techniques. Pharmacological differences were identified between the two NMDA receptor subtypes. The glutamate site antagonist D, L-(epsilon)-2-[3H]amino-4-propyl-5-phosphono-3-pentanoic acid ([3H]CGP 39653) had high affinity for NR1a/NR2A receptors (KD = 3.93 nM) but did not bind to NR1a/NR2B receptors. Glycine site agonists showed a 2.6-5.4-fold higher affinity for NR1a/NR2B receptors. Data from radioligand binding studies indicated that one of the cell lines, NR1a/NR2A-I, expressed a stoichiometric excess of the NR1a subunit, which may exist as homomeric assemblies. This observation has implications when interpreting data from pharmacological analysis of recombinant receptors, as well as understanding the assembly and control of expression of native NMDA receptors.
S Grimwood; B Le Bourdellès; J R Atack; C Barton; W Cockett; S M Cook; E Gilbert; P H Hutson; R M McKernan; J Myers; C I Ragan; P B Wingrove; P J Whiting
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of neurochemistry     Volume:  66     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  1996 Jun 
Date Detail:
Created Date:  1996-07-01     Completed Date:  1996-07-01     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2239-47     Citation Subset:  IM    
Merck, Sharp and Dohme Research Laboratories, Neuroscience Research Centre, Harlow, Essex, England.
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MeSH Terms
2-Amino-5-phosphonovalerate / analogs & derivatives,  metabolism,  pharmacology
Aminoquinolines / metabolism,  pharmacology
Binding Sites / physiology
Blotting, Northern
Cell Line / physiology*
Dizocilpine Maleate / metabolism,  pharmacology
Excitatory Amino Acid Antagonists / metabolism,  pharmacology
Gene Expression / physiology
Glutamine / metabolism,  pharmacology
Glycine / metabolism,  pharmacology
Radioligand Assay
Receptors, N-Methyl-D-Aspartate / agonists,  antagonists & inhibitors,  genetics*
Recombinant Proteins / genetics,  metabolism
Tritium / metabolism
Reg. No./Substance:
0/Aminoquinolines; 0/Excitatory Amino Acid Antagonists; 0/Receptors, N-Methyl-D-Aspartate; 0/Recombinant Proteins; 10028-17-8/Tritium; 132472-31-2/CGP 39653; 139051-78-8/4-trans-2-carboxy-5,7-dichloro-4-phenylaminocarbonylamino-1,2,3,4-tetrahydroquinoline; 56-40-6/Glycine; 56-85-9/Glutamine; 76726-92-6/2-Amino-5-phosphonovalerate; 77086-22-7/Dizocilpine Maleate

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