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Neurochemical, pharmacological, and developmental studies on cerebellar receptors for dicarboxylic amino acids

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Abstract

Specific binding ofl-[3H]glutamate ([3H]Glu) andl-[3H]asparate ([3H]Asp) to cerebellar membranes represented a time-, temperature- pH- and protein-dependent interaction which was both saturable and reversible. Binding sites for both radioligands appeared maximally enriched in synaptosomal fractions isolated by gradient centrifugation. Kinetically derived dissociation constant (K off/K on=K d) for [3H]Glu binding to this fraction indicated high-affinity (443 nM). Competition experiments employing analogs of excitatory amino acids, including new antagonists, helped identify binding sites for [3H]Glu and [3H]Asp as receptors with differential pharmacological, specificities. Membrane freezing reduced numbers of both receptor types, but binding activity could be recovered partially by incubation at 37°C. Glu receptors exhibited a pronounced deleterious sensitivity to thiol modifying reagents andl-Glu (50–1000 μM) provided protection, against these compounds during co-incubation with cerebellar membranes. It is suggested that cold storage may induce partially reversible receptor inactivation by promoting sulfhydryl group/bond modification. Rat cerebellar glutamatergic function (endogenous Glu content, Glu uptake and receptor sites) exhibited an apparent ontogenetic peak between days 8–12 postpartum with a plateauing profile from day 30 to adulthood. The accelerated development (days 8–12) coincides with the first demonstrable Glu release and kainic acid neurotoxicity, as described previously.

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Author notes

  1. N. A. Sharif

    Present address: Department of Biochemistry, University of Nottingham, Queen's Medical Centre, NG7 2UH, Nottingham, UK

Authors and Affiliations

  1. Department of Pharmacology and Experimental Therapeutics, University of Maryland School of Medicine, 21201, Baltimore, Maryland

    N. A. Sharif

  2. Department of Physiology and Pharmacology, Southampton University, S09 3TU, Southampton, UK

    P. J. Roberts

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  1. N. A. Sharif
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Sharif, N.A., Roberts, P.J. Neurochemical, pharmacological, and developmental studies on cerebellar receptors for dicarboxylic amino acids. Neurochem Res 9, 81–101 (1984). https://doi.org/10.1007/BF00967661

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