Toxic Neuronal Death by Glyeraldehyde-3-Phosphate Dehydrogenase and Mitochondria

This proposal was designed to examine the role of glyceraldehyde-3-phosphate dehydrogenase GAPDH, mitochondrial permeability and mitochondrial membrane potential in some forms of toxic neuronal death. Mitochondria have been shown to play a critical decisional role in some forms of cell death, namely apoptotic cell death. Determining the mechanism of the toxin induced apoptosis night reveal potential therapeutic targets which could slow or alleviate the apoptosis. To address these issues, we proposed a series of experiments which include and in vitro study of neurons and neuron-like cells in culture that would be immunostained to determine which toxic insults involved changes in GAPDH levels or distribution, changes mitochondrial membrane potential and permeability transition pore opening. We proposed to examine mutated cell lines of PC12 and 3T3 cells with inducible GAPDH expression. The direct actions of GAPDH and NAD will be examined using a cell free system of nuclear mitochondria and cystolic subfractions. Finally, we proposed to examine whether GAPDH alters the transcription or translation of specific subsets of genes andor their RNA targets using a lysate system. At this point, we addressed many of the objectives and are on schedule for the completion of the study by the end of the funding period.