Ora Goldberg

Barbiturates, alcohols and the CNS excitatory neurotransmission: Specific effects on the kainate and quisqualate receptors

The effects of barbiturates and straight-chain aliphatic alcohols on the responses of rat striatal neurons to excitatory amino acids have been investigated. The responses to N-methyl-D-aspartate, quisqualate, kainate, L-glutamate and L-aspartate were measured by the increase in 22Na+ efflux rate that they produce in brain slices. The responses to quisqualate and kainate, measured in the 22Na+ efflux assay, were found to be partially blocked by barbiturates whereas the responses to N-methyl-D-aspartate, glutamate and aspartate were not. The kainate and quisqualate-induced increases in 22Na+ efflux rate were much more readily blocked by the presence of aliphatic alcohols than were the responses to N-methyl-D-aspartate, glutamate and aspartate. These results strengthen the idea of the existence of 4 distinct receptors for excitatory amino acids in the rat striatum. They are consistent with the presence on the kainate and quisqualate receptors, but not on the N-methyl-D-aspartate and glutamate/aspartate receptors of a hydrophobic domain which would provide a site of interaction for barbiturates and alcohols. They suggest that receptors for excitatory amino acids can be targets for the actions of barbiturates and alcohols on the central nervous system, and may mediate some of the anesthetic and hypnotic effects of these drugs.

An evaluation of selected brain constituents as putative excitatory neurotransmitters

Searching for the endogenous ligands of the 4 classes of excitatory amino acid receptors detected in the mammalian CNS, we have measured, using a 22Na+ efflux receptor assay, the excitatory activity of 42 brain constituents or analogs and established the receptor specificity of those substances which possess excitatory properties. Among the substances tested were methyltetrahydrofolate and N-acetylaspartylglutamate, two putative ligands of the kainate and glutamate receptors. These compounds were found to have very little or no excitatory activity, respectively. The 8 brain constituents possessing excitatory properties displayed a receptor specificity similar to either that of N-methyl-D-aspartate (e.g. quinolinate) or glutamate (e.g. cysteine sulfinate) but not of kainate or quisqualate. These results are discussed in relation with the problem of the identification of brain excitatory neurotransmitters.

The stimulation of ion fluxes in brain slices by glutamate and other excitatory amino acids

SummaryThe stimulation of ion movements by excitatory amino acids in brain slices allows the study of various events related to the process of excitatory neurotransmission. Presynaptic mechanisms of uptake of putative neurotransmitters can be followed by the influx of Na+ ions. Postsynaptic depolarizations due to the activation of action potentials or of ionophores associated with specific receptors can be monitored by measurements of the rate of efflux of radioactive tracer ions. Thus, the pharmacological properties of the excitatory amino acid receptors can be investigated as well as those of their putative endogenous effectors.

Anticonvulsant properties of 3-hydroxy-2-quinoxalinecarboxylic acid, a newly found antagonist of excitatory amino acids

Various aromatic acids have been investigated as putative ligands of excitatory amino acid receptors. 3-Hydroxy-2-quinoxalinecarboxylic acid (HQC), at 1 mM, was found to antagonize the 22Na+ efflux produced in 22Na+-preloaded brain slices by N-methyl-D-aspartate and kainate. The response to glutamate was also affected but not that to quisqualate. The apparent KI value of HQC for suppression of the N-methyl-D-aspartate response was 0.27 mM. The anticonvulsant action of HQC was investigated in mice and rats. HQC administered intracerebroventricularly caused a dose dependent delay in the occurrence of picrotoxin induced convulsions and afforded protection against picrotoxin induced death. These results confirm the proposition that antagonists of excitatory amino acids possess anticonvulsant properties.

Bicyclic lactones derived from kainic acid as novel selective antagonists of neuroexcitatory amino acids.

The bicyclic [2S-(2 alpha,3 beta,4 beta)]-2-carboxy-4-(1-hydroxy-1-methylethyl)-3- pyrrolidineacetic acid delta-lactone (4), as well as its 4-[1-hydroxy-1-(iodomethyl)ethyl], 4-[1-hydroxy-1-(hydroxymethyl)ethyl], and 4-[1-hydroxy-1-[(phenyl-thio)methyl]ethyl] analogues, 6, 7, and 9, respectively, were designed and synthesized as potential selective antagonists of neuroexcitatory amino acids. When applied to rat brain slices, these lactones, which are chemically derived from kainic acid, inhibit the stimulation of Na+ fluxes induced by the neuroexcitants kainic acid and N-methyl-D-aspartic acid. Lactone 4 and the hydroxy lactone 7 block preferentially the response to N-methyl-D-aspartic acid, while the iodo lactone 6 and the phenylthio lactone 9 are mainly kainic acid antagonists. Total inhibitions can be obtained, half of the maximal effect being observed at lactone concentrations in the range of 0.2-3 mM.

Synthesis of novel neuroexcitatory amino acids derived from kainic acid

Abstract The synthesis of the novel epimeric amino acids 5a and 7a by oxidation of methyl ketones derived from kainic acid is described. On biological testing both compounds exhibit a strong neuroexcitatory activity.

Differential sensitivity of selected brain areas to excitatory amino acids

The 22Na+ efflux stimulated by selected agonists of the 4 excitatory amino acid receptors, previously detected in the striatum, has been studied on 22Na+-preloaded slices prepared from 10 major areas of the rat brain. All brain areas were found to be sensitive, albeit to varying extents, to excitatory amino acids. The cerebellum was exceptional in its high sensitivity to kainate and quisqualate and in the absence of effect of N-methyl-D-aspartate. These results support the suggestion that excitatory amino acids interact with heteregenous receptors which differ from each other not only in their pharmacological properties but also in their regional distribution.

Kainyl-bovine serum albumin: a novel ligand of the kainate subtype of glutamate receptor with a very high binding affinity

Bovine serum albumin has been conjugated with kainylaminooxyacetylglycine to afford a multivalent kainylated protein called kainyl-bovine serum albumin (KA-BSA). This derivative, radiolabelled with 125I to more than 5000 Ci/mmol, was found to interact in the chick, goldfish and rat brain to specific membranous sites displaying the pharmacological properties attributed to the kainate sub-type of glutamate receptor. Measurements of the kinetics of association and dissociation of KA-BSA showed a quasi-irreversible binding with dissociation constants in the subpicomolar and nanomolar range. The chemical properties and the binding characteristics of KA-BSA suggest that it interacts mainly with kainate binding sites present in clusters in the membrane. Localization of the KA-BSA binding sites, by autoradiography in the chick cerebellum and by immunoperoxidase staining in the goldfish cerebellum, revealed an exclusive association with the molecular layer.

Lactones derived from kainic acid: Novel selective antagonists of amino acid-induced Na+ fluxes in rat striatum slices

The effects of amino acid lactones chemically derived from the neuroexcitant kainic acid on the response of rat striatal slices to excitatory amino acids, were studied. These compounds antagonize to varying extents the effects of kainic acid and N-methyl-D-aspartic acid but have no effect on the responses to glutamic or quisqualic acid. Some of the lactones antagonize preferentially the effects of kainic acid. This study further confirms the existence of heterogenous populations of excitatory amino acid receptors in the rat striatum.

Peptides derived from kainic acid as antagonists of N-methyl-d-aspartate-induced neuroexcitation in rat brain

The effects of dipeptides and an amide chemically derived from kainic acid (KA) on the response of rat striatal slices to excitatory amino acids were studied. Some of the gamma-peptides of KA were found to antagonize the response to N-methyl-D-aspartate (NMDA) more than that to quisqualate and glutamate and not to have any effect on the response to kainate. The least potent antagonists among the tested compounds were the gamma-amide of KA and the peptides of KA with beta-alanine and gamma-aminobutyric acid, whereas the gamma-kainyl peptides of the alpha-amino acids glycine, tyrosine, glutamate and KA were more active. The latter are the best blockers of the response to NMDA among the tested compounds.