Role of glutathione metabolism in the glutamate-induced programmed cell death of neuronal-like PC12 cells.
In addition to its well-known interaction with ionotropic and metabotropic receptors, glutamate may, at high concentrations, interfere with a cystine-glutamate antiport designated as Xc- and lead to a significant decrease in cystine uptake and intracellular glutathione level. These effects, in turn, may induce death in various cellular bodies including astrocytes, rat glioma cells and cortical neurons in culture. In the present paper we demonstrate that the toxicity evoked by glutamate in a neuronal-like model is indeed related to the metabolism of glutathione since glutamate toxicity is preceded by a significant depletion of intracellular glutathione and is abolished in the presence of precursors of glutathione synthesis such as cystine and N-acetylcysteine. It also appears that prolonged incubation in cystine-free medium leads to cell detachment and death, a phenomenon which is progressively abolished in the presence of increasing concentrations of cystine. In addition, buthionine sulfoximine, a known inhibitor of glutathione synthesis, also induces cell lysis with a time-course very similar to that of glutamate. However, depletion of glutathione is probably not sufficient to trigger the death signal since cycloheximide, which inhibits the toxic effect of both glutamate and buthionine sulfoximine, does not block the decrease in cellular glutathione content induced by these drugs. Our results therefore confirm that oxidative stress and intracellular glutathione depletion are able to trigger programmed cell death in neuronal-like cells, although the exact nature of the death mechanisms remains largely unknown.
Froissard
P
P
Université de Caen, Laboratoire de Neurosciences, CNRS URA 1829, France.
Monrocq
H
H
Duval
D
D
D000111:Acetylcysteine / Q000378:metabolism; D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D016895:Culture Media, Serum-Free; D003513:Cycloheximide / Q000494:pharmacology; D003553:Cystine / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology*; D005978:Glutathione / Q000096:biosynthesis / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000648:ultrastructure; D018384:Oxidative Stress / Q000502:physiology; D016716:PC12 Cells; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats
D000964:Antimetabolites, Antineoplastic; D016895:Culture Media, Serum-Free; D011500:Protein Synthesis Inhibitors; D018698:Glutamic Acid; D003553:Cystine; D019328:Buthionine Sulfoximine; D003513:Cycloheximide; D007770:L-Lactate Dehydrogenase; D005978:Glutathione; D000111:Acetylcysteine