benzyloxycarbonylvalyl-alanyl-aspartyl-fluoromethyl-ketone and Brain-Diseases

The mechanisms underlying kainate (KA) neurotoxicity are still not well understood. We previously reported that KA-mediated neuronal damage in organotypic cultures of hippocampal slices was associated with p53 induction. Recently, both bax and caspase-3 have been demonstrated to be key components of the p53-dependent neuronal death pathway. Caspase activation has also been causally related to the release of mitochondrial cytochrome c (Cyto C) in the cytoplasm as a result of the collapse of the mitochondrial membrane potential (Deltapsi(M)) and the opening of mitochondrial permeability transition pores (mPTP). In the present study, we observed a rapid induction of bax in hippocampal slice cultures after KA treatment. In addition, the levels of Cyto C and caspase-3 were increased in the cytosol while the level of the caspase-9 precursor was decreased. There was also a complete reduction of Rhodamine 123 fluorescence after KA treatment, an indication of Deltapsi(M) dissipation. Furthermore, inhibition of mPTP opening by cyclosporin A partially prevented Cyto C release, caspase activation and neuronal death. These data suggest the involvement of bax, several caspases, as well as Cyto C release in KA-elicited neuronal death. Finally, inhibition of caspase-3 activity by z-VAD-fmk only partially protected neurons from KA toxicity, implying that multiple mechanisms may be involved in KA excitotoxicity.

Topics: Amino Acid Chloromethyl Ketones; Animals; Animals, Newborn; Apoptosis; bcl-2-Associated X Protein; Brain Diseases; Caspase Inhibitors; Caspases; Cells, Cultured; Cyclosporine; Cytochrome c Group; Cytosol; Enzyme Inhibitors; Excitatory Amino Acid Agonists; Hippocampus; Immunohistochemistry; Kainic Acid; Membrane Potentials; Mitochondria; Nerve Degeneration; Neurons; Neuroprotective Agents; Neurotoxins; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Pyramidal Cells; Rats; RNA, Messenger