title: Uptake of L-glutamate into synaptic vesicles: competitive inhibition by dyes with biphenyl and amino- and sulphonic acid-substituted naphthyl groups.
abstract: The specificity of the vesicular L-glutamate carrier was characterized using dyes with biphenyl and amino- and sulphonic acid substituted naphthyl groups, structurally similar to the specific vesicular L-glutamate inhibitor Evans Blue. The dye Trypan Blue was the most potent inhibitor; the IC50 value was determined to be 49 nM. Naphthol Blue Black, Reactive Blue 2, Benzopurpurin 4B, Ponceau SS, Direct Blue 71 and Acid red 114 were also highly potent inhibitors with IC50 values from 330 to 1670 nM (series 1). The dyes were competitive inhibitors of vesicular glutamate uptake, and acted therefore on the glutamate transporter. Their IC50 values for the vesicular uptake of gamma-aminobutyric acid (GABA) were all higher than 20 microM. They had no effect on synaptosomal uptake of glutamate. Furthermore, we have also found several other dyes with IC50 values for the vesicular uptake of glutamate ranging between 1 and 30 microM and for gamma-aminobutyric acid higher than 50 microM (series 2). The most potent inhibitor Trypan Blue contains a biphenyl group, linked by azo groups to side chains containing sulphonic, amino and/or hydroxyl groups coupled to a naphthalene ring system. Trypan Blue and Evans Blue are by molecular mechanics, shown to have planar structures with conjugated double bonds throughout the structure. The other dyes, which were less effective, had phenyl and/or naphthalene groups linked by an azo group. We have also tested a series of amino and/or hydroxyl naphthalene di-/sulphonic acids that correspond to the side chains of the most potent dyes, but they had no effect on glutamate nor on gamma-aminobutyric acid uptake. We conclude that the inhibitory action of these compounds is strictly dependent of the complete molecule.
authors.affiliation: Norwegian Defense Research Establishment, Division for Environmental Toxicology, Kjeller.
authors.forename: S
authors.initials: S
authors.lastname: Roseth
authors.forename: E M
authors.initials: EM
authors.lastname: Fykse
authors.forename: F
authors.initials: F
authors.lastname: Fonnum
mesh_terms: D000818:Animals; D004396:Coloring Agents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005070:Evans Blue / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology*; D018698:Glutamic Acid / Q000378:metabolism*; D008297:Male; D008968:Molecular Conformation; D051381:Rats; D017208:Rats, Wistar; D013329:Structure-Activity Relationship; D013572:Synaptic Vesicles / Q000378:metabolism*; D014343:Trypan Blue / Q000494:pharmacology
chemical_list: D004396:Coloring Agents; D018691:Excitatory Amino Acid Antagonists; D018698:Glutamic Acid; D005070:Evans Blue; D014343:Trypan Blue