Oligodendrocyte precursors and astrocytes in 2-week-old rat primary glial cultures survived 24 h of anoxia, suggesting both cell types could survive using glycolysis for ATP synthesis; however, when the hypoxia developed gradually, the majority of oligodendrocyte precursor cells died within 24 h of the beginning of the experiment but astrocytes survived. Similarly when cultures were exposed to an atmosphere of 1% oxygen, but not 2% or greater, oligodendrocyte precursors died within 24 h. Much more lipid peroxidation was seen under conditions of hypoxia than under conditions of anoxia suggesting that oligodendrocyte precursors died under the former condition because of free radical-induced damage. Using 5-(and -6)-carboxy-2',7'-dichlorodihydrofluorescein (DCFH) as an intracellular probe of oxidative stress, we have demonstrated directly on living cells that oligodendrocyte precursors have a poorer ability to scavenge free radicals than astrocytes. Furthermore, when free radicals were induced to form in the cells either by cysteine auto-oxidation or menadione redox cycling, oligodendrocyte precursors were more readily damaged than astrocytes. We conclude that oligodendroglial precursor cells are exquisitively sensitive to reactive oxygen species.