Fig. 1.

Astrocyte oxygen/glucose deprivation (OGD) paradigm. Rat cortical astrocyte cultures were replated onto 25-mm glass coverslips and maintained for a further 48 h at either 20 or 7% oxygen. Cells were then exposed to OGD in a glucose free ionic shift solution (ISS) with a medium free [O₂] as described in Methods. OGD was terminated by removing the cultures from the anaerobic chamber, replacing the ISS with DMEM/F12 serum free medium containing 5 mM glucose and transferring the cells to incubators at either 7 or 20% O₂. The experiments were terminated at 4 h reoxygenation for measurements of protein and DNA/RNA oxidation, or at 0, 24, or 48 h reoxygenation for measurements of cell death. Control cultures were exposed to serum free medium under 7 or 20% O₂ during the same period as OGD cultures. In some experiments, cells were exposed to only glucose deprivation or O₂ deprivation during the same period as OGD cultures. A–E represents the times when the cells were collected and analyzed.