Neurochemical, pharmacological, and developmental studies on cerebellar receptors for dicarboxylic amino acids.
Specific binding of L-[3H]glutamate ([3H]Glu) and L[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 = Kd) for [3H]Glu binding to this fraction indicated high-affinity (433 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 degrees C. Glu receptors exhibited a pronounced deleterious sensitivity to thiol modifying reagents and L-Glu (50-1000 microM) 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.
D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism*; D002531:Cerebellum / Q000378:metabolism*; D005260:Female; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D051381:Rats; D011919:Rats, Inbred Strains; D017459:Receptors, Amino Acid; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017470:Receptors, Glutamate; D013347:Subcellular Fractions / Q000378:metabolism; D014316:Tritium
D005971:Glutamates; D017459:Receptors, Amino Acid; D011956:Receptors, Cell Surface; D017470:Receptors, Glutamate; C031959:aspartic acid receptor; D014316:Tritium; D001224:Aspartic Acid; D018698:Glutamic Acid