DEATH OF CULTURED POSTNATAL RAT CNS NEURONS BY IN-VITRO HYPOXIA WITH SPECIAL REFERENCE TO N-METHYL-D-ASPARTATE-RELATED TOXICITY

We have established an in vitro hypoxia model using cultured central nervous system neurons from postnatal 4-day-old (P4) rats, in which death may be correlated with N-methyl-D-aspartate (NMDA) -related toxicity. P4 rat hippocampal and neocortical neurons in culture were prevented from death by the addition of MK-801, an NMDA receptor antagonist, and also partially by the removal of calcium ions from the medium, suggesting that NMDA receptors were associated with neuronal death in this in vitro hypoxia model. The neuronal death induced by the model was attenuated by the addition of cr-tocopherol, indicating that free radicals emerged after hypoxia. This event seems similar to the hypoxia-reoxygenation phenomenon in in vitro hypoxia. Continuous treatment with basic fibroblast growth factor (bFGF) during hypoxia, and bFGF pretreatment for 6 h and removal before hypoxia induced the resistance to hypoxia-mediated cell death.