A brief sub-lethal ischaemic stimulus has been reported to protect against subsequent ischaemic damage in vivo, and in vitro following periods of hypoxia or oxygen-glucose deprivation (OGD). Preconditioning against neurotoxic stimuli has been linked to N-methyl-d-aspartate (NMDA) receptors, since receptor blockade prevents the protection afforded by OGD, and low doses of NMDA treatment are capable of preconditioning. The current study demonstrated that NMDA preconditioning also protects against 3-nitropropionic acid (3-NPA), a generator of both excitotoxic and oxidative damage, in addition to glutamate. Cerebellar granule neuronal (CGN) cultures prepared from 8-day neonatal Sprague-Dawley rats were maintained for 8 days prior to NMDA stimulation for 6h. At 9 days in vitro (DIV), preconditioned and control cultures were subjected to a toxic insult (1 microM-10 mM glutamate or 1 microM-10 mM 3-NPA). Neuronal viability was assessed by use of a fluorescein diacetate assay. Protection was effective with 100 microM NMDA preconditioning for 6 h against 1-100 microM glutamate, and also against 1-500 microM 3-NPA. The study demonstrates that NMDA preconditioning can be beneficial against excitotoxic treatments, even when these are potentially complicated by associated oxidative damage and metabolic compromise, as is the case for 3-NPA.