The present study was undertaken to investigate the neuronal protective effect of ceftriaxone (Cef), a β-lactam antibiotic, against delayed neuronal death in hippocampal CA1 subregion normally induced by global brain ischemia in rats, and the role of glial glutamate transporter-1 (GLT-1) in the process. Neuropathological evaluation using thionin staining indicated that pre-administration of Cef once daily for 5 consecutive days significantly prevented the delayed neuronal death in the CA1 subregion normally induced by global brain ischemia in a dose dependent manner, while post-administration of Cef once daily for 5 consecutive days only provided mild neuronal protection against global brain ischemia in the maximal dose. Accompanied with the neuronal protective effect, western blot, immunohistochemistry analysis and ³H-glutamate uptake assay showed that pre-administration of Cef significantly upregulated the expression and glutamate uptake of GLT-1 in hippocampal CA1 subregion. Intracerebroventricular administration of GLT-1 antisense oligodeoxy- nucleotides down-regulating the expression of GLT-1 significantly inhibited the Cef-induced upregualtion in glutamate uptake of GLT-1 and neuronal protective effect against global brain ischemia. Meanwhile, intracerebroventricular administration of dihydrokainate, a selective inhibitor of GLT-1 function, also significantly inhibited the neuronal protective effect of Cef. Thus, it might be concluded that Cef protects neurons against global brain ischemia via upregulating the expression and glutamate uptake of GLT-1.