Glutamate uptake was measured in primary rat cortical astrocyte cultures exposed to sodium azide, 2,4-dinitrophenol, or antimycin A to assess the ability of astrocytes to function under hypoxic conditions. Uptake was maintained at 54-63% of control values despite maximal inhibition of oxidative ATP production. In contrast, the glycolytic inhibitor sodium fluoride (20 mM) reduced glutamate uptake by more than 95% when glucose was the only available substrate. These data suggest that glutamate uptake is largely maintained during hypoxia provided glucose remains available. Astrocyte glutamate uptake may aid neuronal survival during conditions such as incomplete ischemia where oxygen but not glucose is depleted.