In the present study we evaluated a new method to assess the behavioural and biochemical effects of a brief period of acute hypoxia in the brain. In this method, cyanide is injected into the lateral ventricles. Spatial navigation performance in a Morris task was found to be impaired 1 and 5 min after an i.c.v. injection of 5.0 μg cyanide but not after 2.5 μg cyanide. Increased rate of phosphatidic acid formation, reflecting increased phospholipase C activity, were observed after injection of 5.0 μg cyanide, indicating that energy-dependent phosphoinositide metabolism was affected. Chronic treatment with acetyl-carnitine attenuated the cyanide-induced behavioural deficit, but had no effect on energy-dependent phophoinositide metabolism. The results suggest that, in this model, acetyl-l-carnitine may act via free fatty acid metabolism, by increasing the reservoir of activated acyl groups which are involved in the reacylation of membrane phospholipids.