Emerging data indicate that neurotrophic factors and cytokines utilize similar signal transduction mechanisms. Although neurotrophic factors can protect CNS neurons against a variety of insults, the role of cytokines in the injury response is unclear. We now report that TNFβ and TNFα (1–100 ng/ml) can protect cultured embryonic rat hippocampal, septal, and cortical neurons against glucose deprivation-induced injury and excitatory amino acid toxicity. The elevation of intracellular calcium concentration ([Ca²⁺]_i),) induced by glucose deprivation, glutamate, NMDA, or AMPA was attenuated in neurons pretreated with TNFβ. The mechanism whereby TNFs stabilize [Ca²⁺]_i may involve regulation of the expression of proteins involved in maintaining [Ca²⁺]_i homeostasis, since both TNFβ and TNFα caused a 4- to 8-fold increase in the number of neurons expressing the calcium-binding protein calbindin-D_28k. These data suggest a neuroprotective role for TNFs in the brain's response to injury.