Pathological activation of microglia, which reside quiescently in physiological CNS, is associated with various neurodegenerative diseases. Endogenous polyamines, spermidine and spermine, are known to be activators of cell proliferation and differentiation. We previously reported that both spermidine and spermine induce dose-dependent cell death in cultured rat brain microglia at a submicromolar concentration range via apoptotic process, whereas cultured astrocytes were less sensitive to these polyamines [Neuroscience 120 (2003) 961]. These polyamine effects were observed only in the presence of fetal bovine serum. In the present study we examined further the mechanism of polyamine-induced microglial cell death. Amine oxidase in fetal bovine serum produces hydrogen peroxide and an aminoaldehyde from spermine, and the latter generates acrolein spontaneously. Acrolein was found to be much more toxic to microglia than to astrocytes and the effective concentration of acrolein was similar to that of spermine, whereas hydrogen peroxide was marginally toxic. Aminoguanidine, an inhibitor of amine oxidase, blocked the toxic effects of spermine on microglia. Spermine cytotoxicity was also prevented by antioxidant reagents; glutathione (reduced form), cysteine, and N-acetylcysteine. These results suggest that polyamine-induced apoptotic cell death of microglia is triggered by an oxidative stress with acrolein, which is produced by amine oxidase from polyamine. The different toxicities of polyamine between two glial cells may regulate the balance of glial activation in some pathological conditions of CNS.