A rapid filtration assay for the glycine binding site on the NMDA receptor in rat cortical membranes using [3H]dichlorokynurenic acid.
A filtration binding assay using [3H]dichlorokynurenic acid to label the glycine binding site on the N-methyl-D-aspartic acid receptor has been evaluated on rat cortical membranes. This ligand binds to a single population of binding sites following mass action kinetics with a KD of 29 nM and a capacity of 5.73 pmol (mg protein)-1. The pharmacological specificity of the binding site is identical to that previously reported for this binding site using [3H]glycine as a radioligand. Agonists showed lower affinity and antagonists higher affinity when [3H]dichlorokynurenic acid was used compared with [3H]glycine. The higher affinity of [3H]dichlorokynurenic acid compared with [3H]glycine make it the more suitable compound with which to label the glycine site.
Canton
T
T
Biology Department, Rhône-Poulenc Rorer, Centre de Recherches de Vitry, Vitry-sur-Seine, France.
Doble
A
A
Miquet
J M
JM
Jimonet
P
P
Blanchard
J C
JC
D000818:Animals; D001665:Binding Sites; D002540:Cerebral Cortex / Q000378:metabolism*; D005374:Filtration; D005998:Glycine / Q000378:metabolism*; D066298:In Vitro Techniques; D007736:Kynurenic Acid / Q000031:analogs & derivatives*; D008297:Male; D011869:Radioligand Assay; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000737:chemistry*; D014316:Tritium
D016194:Receptors, N-Methyl-D-Aspartate; D014316:Tritium; D007736:Kynurenic Acid; C066192:5,7-dichlorokynurenic acid; D005998:Glycine