Reducing Conditions Produce a Loss of Neuroprotective Efficacy of Competitive but not non-Competitive Antagonists in a Model of NMDA-Mediated Excitotoxicity in Organotypic Hippocampal Slice Cultures

Abstract

Experimental data indicate that NMDA receptor activation is strongly implicated in the pathogenesis of cerebral ischaemia. However, the results from in vivo studies are equivocal, with NMDA antagonists being active in only some models. It has recently been demonstrated that competitive and non-competitive NMDA antagonists behave differently under normal and ischaemic conditions. These studies have used organotypic hippocampal slice cultures to investigate whether this disparity is due to redox-modulation of the NMDA receptor which occurs in ischaemia. NMDA-mediated toxicity was concentration dependent with little damage occurring with less than 10 µM NMDA and maximal damage produced by 30 µM. NMDA toxicity was significantly enhanced by pre-treatment with 1 mM dithiothreitol, a reducing agent, such that damage occurred at 1 nM NMDA, and maximal damage was produced by 10 µM. The efficacy of MK-801 was not altered by reducing conditions, but the EC50 of the competitive antagonist APV was increased by 20-fold. These data strongly suggest that the neuroprotective efficacy of NMDA antagonists is significantly altered under ischaemic conditions, and that more beneficial effects will be obtained with antagonists having a higher affinity for the receptor in the reduced configuration.