title: Mechanisms of oxygen glucose deprivation-induced glutamate release from cerebrocortical slice cultures.
abstract: Glutamate has been recognized to mediate ischemia-induced neuronal injury in the brain, but the source of extracellular glutamate during ischemic insults remains controversial. We investigated the mechanisms of glutamate release in organotypic cerebrocortical slice cultures prepared from rat neonates, using oxygen glucose deprivation (OGD) as an in vitro ischemia model. Slice cultures were submerged in glucose-free deoxygenated buffer for 20-60 min and glutamate released into the extracellular buffer was quantified. Cell injury was assessed by uptake of propidium iodide 24 h after OGD insult. OGD-induced time-dependent glutamate release and cell injury, both of which were potently inhibited by a sodium channel blocker tetrodotoxin (1 microM). Application of voltage-dependent Ca2+ channel blockers or of an inhibitor of vacuolar-ATPase significantly reduced OGD-induced glutamate release and cell injury. On the contrary, inhibitors of glutamate transporters exacerbated OGD-induced glutamate release and cell injury. Volume sensitive organic anion channel blockers also augmented OGD-induced glutamate release and cell injury. In addition, OGD-induced glutamate release was markedly reduced in neuron-depleted slice cultures that were pretreated with 100 microM NMDA. These results suggest that vesicular release of neuronal origin constitutes a crucial component of extracellular glutamate increase during ischemic insults, which triggers neuronal injury.
authors.affiliation: Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, 46-29 Yoshida-shimoadachi-cho, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Fujimoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Katsuki
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Kume
authors.forename: Shuji
authors.initials: S
authors.lastname: Kaneko
authors.forename: Akinori
authors.initials: A
authors.lastname: Akaike
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000378:metabolism*; D002545:Brain Ischemia / Q000378:metabolism / Q000503:physiopathology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology*; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005947:Glucose / Q000172:deficiency*; D018698:Glutamic Acid / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D009924:Organ Culture Techniques; D010100:Oxygen / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism
chemical_list: D002121:Calcium Channel Blockers; D004791:Enzyme Inhibitors; D018691:Excitatory Amino Acid Antagonists; D016194:Receptors, N-Methyl-D-Aspartate; D018698:Glutamic Acid; D005947:Glucose; D010100:Oxygen