Lipoxygenase Metabolism of Arachidonic Acid in Brain

Abstract: When blood‐free mouse brain slices were incubated with exogenous radiolabeled arachidonic acid, gas chromatography/mass spectrometry confirmed that the major radioactive lipoxygenase enzyme product of arachidonic acid was 12‐hydroxy‐5,8,10,14‐eicosatetraenoic acid (12‐HETE), with lesser amounts of 5‐hydroxy‐6,8,11,14‐eicosatetraenoic acid and 15‐hydroxy‐5,8,11,13‐eicosatetraenoic acid. When 12‐[2H]HETE was used to measure endogenous 12‐HETE in brain tissue frozen with liquid nitrogen, the level of 12‐HETE was 41 ± 6 ng/g of wet weight tissue. This frozen tissue level was not due to the presence of blood. When brain slices were incubated in vitro for 20 min, the 12‐HETE level increased to 964 ± 35 ng/g of wet weight tissue. Elimination of residual intravascular blood before tissue incubation reduced the brain slice 12‐HETE concentration by one‐half.

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