title: Endobain E, a brain endogenous factor, is present and modulates NMDA receptor in ischemic conditions.
abstract: We have isolated from rat cerebral cortex an endogenous Na(+), K(+)-ATPase inhibitor, termed endobain E, which modulates glutamatergic N-methyl-d-aspartate (NMDA) receptor. This endogenous factor allosterically decreases [(3)H]dizocilpine binding to NMDA receptor, most likely acting as a weak channel blocker. In the present study we investigated whether endobain E is present in the cerebral cortex of rats subjected to ischemia and modulates NMDA receptor exposed to the same conditions. Ischemia-reperfusion was carried out by bilateral occlusion of common carotid arteries followed by a 15-min reperfusion period. Elution profile of brain soluble fraction showed that endobain E is present in cerebral cortex of ischemia-reperfusion rats. On assaying its effect on synaptosomal membrane Na(+), K(+)-ATPase activity and [(3)H]dizocilpine binding to cerebral cortex membranes prepared from animals without treatment, it was found that the endogenous modulator isolated from ischemia-reperfusion rats was able to inhibit both enzyme activity and ligand binding. On the other hand, endobain E prepared from rats without treatment also decreased binding to cerebral cortex or hippocampal membranes obtained from animals exposed to ischemia-reperfusion. Since ischemia decreases tissue pH and NMDA receptor activity varies according to proton concentration, pH influence on endobain E effect was tested. Endobain E ( approximately 80 mg original tissue) decreased [(3)H]dizocilpine binding 25% at pH 7.4 or 8.0 but 90% at pH 6.5. These results demonstrate that endobain E is present and also able to modulate NMDA receptor in the short-term period that follows cerebral ischemia and that its effect depends on proton concentration, suggesting greater NMDA receptor modulation by endobain E at low pH, typical of ischemic tissues.
authors.affiliation: Instituto de Biología Celular y Neurociencias "Prof. E. De Robertis", Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: A
authors.initials: A
authors.lastname: Reinés
authors.forename: S
authors.initials: S
authors.lastname: Zárate
authors.forename: C
authors.initials: C
authors.lastname: Carmona
authors.forename: G
authors.initials: G
authors.lastname: Negri
authors.forename: C
authors.initials: C
authors.lastname: Peña
authors.forename: G
authors.initials: G
authors.lastname: Rodríguez de Lores Arnaiz
mesh_terms: D000818:Animals; D002545:Brain Ischemia / Q000150:complications / Q000201:enzymology / Q000378:metabolism*; D002540:Cerebral Cortex / Q000201:enzymology / Q000378:metabolism*; D004195:Disease Models, Animal; D016291:Dizocilpine Maleate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000201:enzymology / Q000378:metabolism; D008297:Male; D010042:Ouabain / Q000031:analogs & derivatives* / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D015427:Reperfusion Injury / Q000201:enzymology / Q000209:etiology / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology / Q000378:metabolism
chemical_list: D016194:Receptors, N-Methyl-D-Aspartate; C119205:endobain E; D010042:Ouabain; D016291:Dizocilpine Maleate; D000254:Sodium-Potassium-Exchanging ATPase