A detailed analysis of hydrogen peroxide-induced cell death in primary neuronal culture

A variety of neurodegenerative disease states have been associated with oxidative damage or stress. Such stress is thought to be mediated by excessive exposure of cells to reactive oxygen species such as free radicals, which can be generated following cell lysis, oxidative burst (as part of the immu...

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Bibliographic Details
Published in:Neuroscience 1995-08, Vol.67 (4), p.921-932
Main Authors: Whittemore, E.R., Loo, D.T., Watt, J.A., Cotmans, C.W.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Calcium - metabolism
Cell Death
Cells, Cultured - drug effects
DNA Probes
Fundamental and applied biological sciences. Psychology
Fura-2
Hydrogen Peroxide - pharmacology
Isolated neuron and nerve. Neuroglia
Microscopy, Electron
Models, Neurological
Neurons - drug effects
Rats
Rats, Inbred Strains
Time Factors
Vertebrates: nervous system and sense organs
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Summary:A variety of neurodegenerative disease states have been associated with oxidative damage or stress. Such stress is thought to be mediated by excessive exposure of cells to reactive oxygen species such as free radicals, which can be generated following cell lysis, oxidative burst (as part of the immune response) or by the presence of an excess of free transition metals. Since the neuronal death observed in neurodegenerative disease may be related to free radical damage, we were interested in developing a model system to investigate the mechanisms by which reactive oxygen species may damage or kill neurons. To this end, we have recently reported that brief exposure of cultured cortical neurons to H 2O 2 can induce neuronal death that proceeds via an apoptotic cell suicide pathway. The studies reported here investigate H 2O 2-induced cell death in more detail. Our data suggest that exposure of cultured cortical neurons to H 2O 2 can induce apoptotic cell death within 3 h, as assessed by cell viability, morphological and ultrastructural measures. In addition, experiments presented show that exposure to high concentrations of H 2O 2 (100 μM) causes increases in intracellular free calcium within 3 h, suggesting that increased intracellular calcium may be associated with some aspects of H 2O 2-induced cell death. However, at intermediate concentrations of H 2O 2 (30 μM), intracellular calcium remained stable during a 3 h exposure, during which time membrane blebbing was observed in ultrastructural studies. This suggests that some aspects of apoptotic cell death induced by H 2O 2 may not be associated with increased intracellular free calcium. Thus, this model appears valuable for studies of the mechanism(s) by which oxidative injury may induce apoptotic cell death and damage to neurons in the CNS.
ISSN:0306-4522
1873-7544
DOI:10.1016/0306-4522(95)00108-U