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GLUTAMATE-DEHYDROGENASE IMPROVES BINDING OF [H-3] CGP39653 TO NMDA RECEPTORS IN THE AUTORADIOGRAPHIC ASSAY

JAARSMA, D., SEBENS, JB. & KORF, J., Feb-1993, In : Journal of Neuroscience Methods. 46, 2, p. 133-138 6 p.

Research output: Contribution to journalArticleAcademicpeer-review

The novel high-affinity competitive NMDA receptor antagonist, CGP39653, is employed as radioligand to autoradiographically label the NMDA receptor in rat and human brain. Glutamate dehydrogenase (GlDH; E.C.1.4.1.3) was added to the incubation buffer to degrade residual endogenous L-glutamate, which was not entirely removed from the section after the prewashing step and interferred with [H-3]CGP39653 binding. At 20 nM [H-3]CGP39653, GlDH increased specific binding as much as 5 times, depending on the dose of GlDH and the presence of NAD+, and hydrazine. Scatchard plots of binding data revealed that this increase was due to a decrease of the K(D) from 148 nM to 33 nM in the absence and the presence of GlDH, respectively. Addition of GlDH in the NMDA receptor autoradiographic assay may be of importance in quantitative studies with human brain tissue which may contain variable levels of endogenous L-glutamate.

Original languageEnglish
Pages (from-to)133-138
Number of pages6
JournalJournal of Neuroscience Methods
Volume46
Issue number2
Publication statusPublished - Feb-1993

    Keywords

  • [H-3]CGP39653, CNQX, NMDA RECEPTOR, AMPA RECEPTOR, RECEPTOR AUTORADIOGRAPHY, EXCITATORY AMINO ACID, AMINO-ACID RECEPTORS, D-ASPARTATE, RAT-BRAIN, KAINIC ACID, SITES, MEMBRANES, AMPA

ID: 6342588