Ischemia in neuronal PC12 cells, role of free radical generation and cellular damage

grantor: University of TorontoOxidative stress causing impaired cell function underlies many neurodegenerative disorders. I used an in-vitro model, neuronal PC12 cells to study the effects of Oxygen-Glucose-Deprivation (OGD/GD/OD). Cytotoxicity in culture medium (DNEM) was significantly higher following OGD/GD, immediately and 24hrs later, than the typically used--balanced salt solution (BSS). Cell death significantly decreased with addition of a metabolic substrate, inhibition of glycolysis significantly increased cell death. Free radical generation (ROS) studied with 5'6'CMH2DCF-DA, using confocal-microscopy and flow-cytometry, showed a significant early and persistent increase during OGD and early reperfusion, in DMEM. Lipid-peroxidation (TBARS assay) was increased in DMEM following OGD. Changes in mitochondrial morphology during OGD, coincided with the rise in ROS. NGF, hypothermia and anti-oxidants improved cell survival. These results indicate that oxidative stress may be an important mediator of cytotoxicity in neuronal PC12 cells during OGD in DMEM. Neuroprotective treatments must target strategies to decrease oxidative stress in the brain.M.Sc