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We here demonstrate that general anesthesia with isoflurane can have profound effects on the brain of mice long after the anesthetic has been discontinued. Three hours of exposure to 1% isoflurane induced rapid and longlasting protection against 60 min transient focal cerebral ischemia induced by filament occlusion of the middle cerebral artery (MCAO). Mean infarct volumes were significantly smaller in animals pretreated with isoflurane 0, 12, and 24 h before MCAO (−38%, −31%, −24%, respectively). Mild hypoxia (17% O2) during or 5 mg/kg desferrioxiamine administered at the onset of isoflurane pretreatment completely abrogated the development of delayed tolerance (12 h) against focal cerebral ischemia, suggesting that the signaling of delayed protection induced by isoflurane is sensitive to the intracellular oxygenation state.