title: Effects of thiocyanate and AMPA receptor ligands on (S)-5-fluorowillardiine, (S)-AMPA and (R,S)-AMPA binding.
abstract: AMPA receptors can be labeled using the agonist radioligands [3H](R,S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid ([3H](R,S)-AMPA), [3H](S)-AMPA or [3H](S)-5-fluorowillardiine. In the presence of KSCN, [3H](R,S)-AMPA and [3H](S)-AMPA bind to a single population of sites in rat brain membranes, whereas [3H](S)-5-fluorowillardiine binds with two affinity components. KSCN increased the affinity of the low affinity [3H](S)-5-fluorowillardiine component > 4-fold and increased the density of both components 1.5-1.7-fold, arguing against KSCN-induced interconversion of low to high affinity states. KSCN, which promotes receptor desensitization, increased the potency of AMPA isomers, (S)-5-fluorowillardiine, quisqualate and cyclothiazide for inhibition of [3H](S)-5-fluorowillardiine binding suggesting that these ligands discriminate desensitized and nondesensitized receptors. In contrast, KSCN did not greatly affect the potency of glutamate, kainate, or competitive antagonists suggesting that these ligands do not discriminate desensitized and nondesensitized receptors. In the presence of KSCN, the rank order potency for agonists and antagonists was similar or identical in all assays indicating that the three radioligands bind identical glutamate recognition sites, a conclusion supported by their identical total receptor density. However, AMPA isomers displayed 6-10-fold higher potency for displacement of [3H](S)- or (R,S)-AMPA relative to [3H](S)-5-fluorowillardiine binding. This finding, coupled with the marked two component binding by [3H](S)-5-fluorowillardiine but not [3H](S)- or (R,S)-AMPA, suggests qualitative differences between the interaction of these ligands with the agonist recognition site.
authors.affiliation: CoCensys Inc., Irvine, CA 92618, USA.
authors.forename: J E
authors.initials: JE
authors.lastname: Hawkinson
authors.forename: S A
authors.initials: SA
authors.lastname: Espitia
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives* / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D018690:Excitatory Amino Acid Agonists / Q000378:metabolism*; D066298:In Vitro Techniques; D008024:Ligands; D008297:Male; D011743:Pyrimidines / Q000378:metabolism*; D051381:Rats; D018091:Receptors, AMPA / Q000187:drug effects* / Q000378:metabolism; D013237:Stereoisomerism; D013861:Thiocyanates / Q000494:pharmacology*; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000378:metabolism*
chemical_list: D018690:Excitatory Amino Acid Agonists; D008024:Ligands; D011743:Pyrimidines; D018091:Receptors, AMPA; D013861:Thiocyanates; C095178:5-fluorowillardiine; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; D000409:Alanine; C009941:potassium thiocyanate