Please use this identifier to cite or link to this item: http://ahro.austin.org.au/austinjspui/handle/1/12631
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dc.contributor.authorCincotta, Men
dc.contributor.authorSummers, R Jen
dc.contributor.authorBeart, P Men
dc.date.accessioned2015-05-16T02:21:25Z
dc.date.available2015-05-16T02:21:25Z
dc.date.issued1989-02-15en
dc.identifier.citationAnalytical Biochemistry; 177(1): 150-5en
dc.identifier.govdoc2568102en
dc.identifier.otherPUBMEDen
dc.identifier.urihttp://ahro.austin.org.au/austinjspui/handle/1/12631en
dc.description.abstractA method was developed for radiolabeling excitatory amino acid receptors of rat brain with L-[3H]glutamate. Effective labeling of glutamate receptors in slide-mounted 10-microns sections was obtained using a low incubation volume (0.15 ml) and rapid washing: a procedure where high ligand concentrations were achieved with minimal waste. Saturation experiments using [3H]glutamate revealed a single binding site of micromolar affinity. The Bmax was trebled in the presence of Ca2+ (2.5 mM) and Cl- (20 mM) with no change in the Kd. Binding was rapid, saturable, stereospecific, and sensitive to glutamate receptor agonists. The proportions of [3H]glutamate binding sensitive to N-methyl-D-aspartate (NMDA), kainate, and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) were 34, 54, and 51%, respectively. NMDA inhibited binding at a distinct subset of L-[3H]glutamate sites, whereas AMPA and kainate competed for some common sites. Labeling of sections with L-[3H]glutamate in the presence of the selective agonists allowed autoradiographic visualization of glutamate receptor subtypes in brain tissue.en
dc.language.isoenen
dc.subject.otherAnimalsen
dc.subject.otherAspartic Acid.analogs & derivatives.metabolismen
dc.subject.otherAutoradiographyen
dc.subject.otherBinding, Competitiveen
dc.subject.otherBrain.metabolismen
dc.subject.otherGlutamates.diagnostic useen
dc.subject.otherGlutamic Aciden
dc.subject.otherIbotenic Acid.analogs & derivatives.metabolismen
dc.subject.otherKainic Acid.metabolismen
dc.subject.otherKineticsen
dc.subject.otherMaleen
dc.subject.otherN-Methylaspartateen
dc.subject.otherRadioligand Assayen
dc.subject.otherRatsen
dc.subject.otherRats, Inbred Strainsen
dc.subject.otherReceptors, Glutamateen
dc.subject.otherReceptors, Neurotransmitter.analysisen
dc.subject.otherSoftwareen
dc.subject.otherTritium.diagnostic useen
dc.subject.otheralpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Aciden
dc.titleBiochemical characterization of an autoradiographic method for studying excitatory amino acid receptors using L-[3H]glutamate.en
dc.typeJournal Articleen
dc.identifier.journaltitleAnalytical biochemistryen
dc.identifier.affiliationUniversity of Melbourne, Clinical Pharmacology and Therapeutics Unit, Austin Hospital, Heidelberg, Victoria, Australiaen
dc.description.pages150-5en
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pubmed/2568102en
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