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Our previous study showed that 3-min cerebral ischemic preconditioning (CIP) up-regulated the expression of glial glutamate transporter-1 (GLT-1) protein, which protects pyramidal neurons, allowing them to survive an 8-min ischemic insult that usually induces severe delayed neuronal death in the hippocampal CA1 subfield. In the present study, in situ hybridization and immunohistochemistry were used to observe whether GLT-1 mRNA is modulated and whether actrocytes and/or neurons express GLT-1 mRNA during the induction of brain ischemic tolerance. We observed that GLT-1 mRNA is expressed in neurons and astrocytes in the hippocampal CA1 subfield. The expression of GLT-1 mRNA was significantly down-regulated in both neurons and astrocytes after the 8-min lethal ischemic insult. CIP for 3 min increased the expression of GLT-1 mRNA in neurons and astrocytes, and induced the elongation of the astrocytic processes around pyramidal neurons. It may be concluded that CIP performed 2 days before lethal ischemic insult activated astrocytes, which resulted in an increased number of lengthened processes expressing high levels of GLT-1, which protected the neurons and allowed them to survive 8-min ischemic insult that is usually lethal to neurons in the hippocampal CA1 subfield.