Intracellular generation of free radicals and modifications of detoxifying enzymes in cultured neurons from the developing rat forebrain in response to transient hypoxia
Section snippets
Neuronal cell culture and exposure to hypoxia
Animal experimentation was carried out with the highest standards of animal care and housing, according to the N.I.H. Guide for the Care and Use of Laboratory Animals. Primary cultured neurons were obtained from 14-day-old rat embryo forebrains as previously described (Chihab et al., 1998a, Chihab et al., 1998b, Bossenmeyer-Pourié et al., 1999b, Bossenmeyer-Pourié et al., 2000b, Lièvre et al., 2000). When they were in the proestrus period, as shown by the observation of daily vaginal smears,
Neuronal injury
By using antibodies raised against neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP), immunohistochemical characterization of culture preparations grown for 6 days in chemically defined medium confirmed the high proportion of differentiated neuronal cells, showing 92.5±2.8% of NSE-positive cells, with 7.6±1.6% of GFAP-positive cells (data not shown).
Transient exposure to the anaerobic gas mixture usually reduced the partial pressure of oxygen (PO2) in the culture medium
Discussion
Due to its metabolic properties, oxidative capacities and specific composition, the brain is particularly sensitive to oxidative damage. Within the CNS, the response to oxidative stress has been shown to vary according to the various cellular phenotypes, and the antioxidant capacity of neurons is lower than that of glia (Makar et al., 1994). Furthermore, it has been reported that the immature brain exhibits reduced defense mechanisms against oxidative events (Mishra and Delivoria-Papadopoulos,
Acknowledgements
V.L. wishes to express her gratitude to Mrs. Catherine Charoy for her generous support.
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