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Damage to central nervous system white matter is observed following hypoglycemia, raising the possibility that hypoglycemia influences oligodendrocytes and myelination. To examine effects of hypoglycemia on oligodendrocytes and myelin formation, we studied cultured oligodendrocyte precursor cells and cerebellar slice cultures. We observed that with decreasing concentrations of glucose, oligodendrocyte precursor cell proliferation, maturation, and migration decreased. We also observed that hypoglycemia induced apoptotic cell death and activation of caspase-3 in oligodendrocyte precursor cells. Slice culture studies showed that glucose is required for myelinated fiber formation, as with reduction in the glucose concentration, the density of myelinated fibers decreased. Collectively, these data show that hypoglycemia inhibits oligodendrocyte development and myelination and that hypoglycemia triggers apoptotic cell death in oligodendrocyte precursor cells.