We investigated the effect of glutathione (GSH) depletion on mitochondrial function and generation of reactive oxygen intermediates (ROI) in PC12 cells in vitro. Direct depletion of cellular GSH using ethacrynic acid (EA, 500 mM) resulted in a concentration-dependent generation of ROI and cell death within 24 h. Treatment with 500 μ M l -buthionine sulfoximine (BSO), which inhibits GSH synthesis, reduced cellular GSH but did not lead to generation of ROI. Furthermore, cells remained viable up to 72 h. Analysis of subcellular fractions revealed complete loss of cytosolic and mitochondrial GSH within 4 h of EA treatment. In contrast, BSO-treated cells still maintained 100% GSH in the mitochondrial fraction for 4 h and 6% for 48 h. Mitochondrial complex II/III and IV activities were not significantly decreased up to 48 h of BSO treatment while EA treatment resulted in a complete loss of complex II/III activity and a 70% reduction of complex IV activity within 4 h. These findings suggest that mitochondrial GSH is critical for the maintenance of mitochondrial function and cellular viability.
Neuroscience Letters – Elsevier
Published: Apr 2, 1999
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