Chloride-dependent acute excitotoxicity in adult rat retinal ganglion cells

Mechanisms of excitotoxic degeneration of retinal ganglion cells (RGCs) remain controversial, due to the lack of suitable in vitro experimental systems for evaluation of RGC death. In this study, we investigated acute excitotoxicity in RGCs using eyecup preparations obtained from adult rats, with special reference to ionic dependence of N-methyl-D-aspartate (NMDA) and kainate toxicity. Retrograde labeling of RGCs with a fluorescent tracer diamidino yellow, combined with labeling of dead cells by propidium iodide, enabled us to discriminate dead RGCs from other cells in the ganglion cell layer. Exposure of eyecups to NMDA or kainate for 30min followed by 6h post-incubation caused cell death in a subpopulation of RGCs as well as other (presumably displaced amacrine) cells. RGCs in the peripheral area of the retina were less sensitive to NMDA toxicity than those in the central area. Death of RGCs and other retinal cells by NMDA or kainate was largely abolished by substitution of extracellular Cl(-), whereas chelation of extracellular Ca(2+) did not inhibit NMDA or kainate toxicity in RGCs. Strychnine but not bicuculline partially inhibited NMDA-induced RGC death, although these drugs were not effective against kainate-induced RGC death. On the other hand, niflumic acid, a Cl(-) channel blocker, markedly inhibited RGC death induced by kainate as well as by NMDA. These results underscore the important role of Cl(-) in acute excitotoxicity in adult rat RGCs.