The present study was designed to determine whether the inhibition of glutamate release in organum vasculosum laminae terminalis (OVLT) of rabbit brain by acetaminophen might be protective in a whole-animal model of staphylococcal enterotoxin A (SEA) fever. Unanesthetized rabbits were administered intravenously with SEA, and both colonic temperature (Tco) and glutamate release in OVLT were measured simultaneously. The glutamate release in OVLT was measured with a microdialysis probe previously implanted. Both the Tco and glutamate release in OVLT were simultaneously increased following intravenous administration of SEA. The SEA-induced rise in both the Tco and the levels of glutamate release in OVLT were suppressed by pretreatment with intravenous injection of acetaminophen (1, 5 or 10 mg/kg). Furthermore, treatment of OVLT with acetaminophen (50-150 microg) attenuated the fever-like hyperthermia induced by intra-OVLT injection of glutamate. Our results show acetaminophen may reduce glutamate release in OVLT and result in antipyresis.