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Pharmacological characterization of the N-methyl-d-aspartate (NMDA) receptor-channel in rodent and dog brain and rat spinal cord using [3H]MK-801 binding

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Abstract

The biochemical and pharmacological properties of [3H]MK-801 binding to the N-methyl-d-aspartate (NMDA) receptor-channel in homogenates of mouse, guinea pig and dog brain, dog cerebral cortex and rat spinal cord were determined using radioligand binding techniques. Specific [3H]MK-801 binding increased linearily with increasing tissue concentration and in general represented 80–93% of the total binding at 6–8 nM radioligand concentration. [3H]MK-801 interacted with brain and spinal homogenates with high affinity. The dissociation constants (K d ) for all tissues studied were similar ranging between 7.9 and 11.9 nM, whereas the maximum number of binding sites (Bmax) showed a wide, tissue-dependent range (0.1–6.75 pmol/mg protein). The rank order of tissue enrichment was found to be as follows: mouse brain>>dog cerebral cortex>>dog brain>> guinea pig brain>>rat spinal cord. Specific [3H]MK-801 binding in rodent and dog brain, dog cerebral cortex and rat spinal cord exhibited a similar pharmacological profile 9correlation coefficients=0.93–0.99). The rank order of potency of unlabelled compounds competing for [3H]MK-801 binding was: (+)MK-801>(−)MK-801>phencyclidine>(−)cyclazocine>>(+)cyclazocine ≥ ketamine>(+)N-allyl-N-normetazocine>(−)N-allyl-N-normetazocine>(−)pentazocine>(+)pentazocine. NMDA, Kainate, quisqualate and several other compounds failed to inhibit [3H]MK-801 binding at 100 μM. In modulation studies conducted on extensively washed dog cortex membranes, Mg2+ ions stimulated [3H]MK-801 binding at 10 μM-1 mM (EC50=91.5 μM) and then inhibited the binding from 1 mM to 10 mM (IC50=3.1 mM). Glycine stimulated [3H]MK-801 binding at 30 nM-1 mM (EC50=256 nM). In contrast, Zn2+ ions inhibited the binding of [3H]MK-801 binding site exhibited similar pharmacological and biochemical properties. These data appear to suggest that the pharmacological profile of the NMDA-receptor-channel is species and tissue independent.

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Authors and Affiliations

  1. Department of Neuroscience, Institute of Pharmacology (R7-101), Syntex Research, 3401 Hillview Avenue, 94303, Palo Alto, CA

    N. A. Sharif, J. L. Nunes & R. L. Whiting

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Sharif, N.A., Nunes, J.L. & Whiting, R.L. Pharmacological characterization of the N-methyl-d-aspartate (NMDA) receptor-channel in rodent and dog brain and rat spinal cord using [3H]MK-801 binding. Neurochem Res 16, 563–569 (1991). https://doi.org/10.1007/BF00974875

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