title: [3H](+)MK801 radioligand binding assay at the N-methyl-D-aspartate receptor.
abstract: The N-methyl-D-aspartate subtype of glutamate receptor is unusual in that it requires two endogenous agonists for activation. Thus, in addition to glutamate, the amino acid glycine (or possibly D-serine) is an essential co-agonist. This unit presents a radioligand binding protocol that detects ligand activity at the NMDA receptor-associated glycine site. This is a convenient approach that exploits the ability of NMDA receptor modulators to alter the kinetics of ligands that bind to the channel-blocking site of the NMDA receptor. This protocol takes advantage of one of the most potent and specific ligands that bind to this receptor, in this case [(3)H](+)MK801. Importantly, this assay can detect and differentiate agonists and antagonists that bind to the glycine site. A protocol for the measurement of glycine site activity with [(3)H](+)MK801 binding is provided, along with support protocols that provide information to aid in the design of assays of agonists and antagonists of the glycine site, and data analysis.
authors.affiliation: University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: I J
authors.initials: IJ
authors.lastname: Reynolds
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D016291:Dizocilpine Maleate / Q000494:pharmacology*; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology*; D008566:Membranes / Q000378:metabolism; D011869:Radioligand Assay / Q000379:methods*; D051381:Rats; D018009:Receptors, Glycine / Q000819:agonists / Q000037:antagonists & inhibitors; D016194:Receptors, N-Methyl-D-Aspartate / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013223:Statistics as Topic; D014316:Tritium
chemical_list: D018691:Excitatory Amino Acid Antagonists; D018009:Receptors, Glycine; D016194:Receptors, N-Methyl-D-Aspartate; D014316:Tritium; D016291:Dizocilpine Maleate