Incorporation of arachidonic and docosahexaenoic acids into phospholipids of rat brain membranes

Abstract

The incorporation of [ 3H]arachidonic acid (20:4n-6) into rat brain membranes and its mobilization in response to norepinephrine, a relevant neuromediator were studied. The most efficient [ 3H]20:4n-6 incorporation was in inositol glycerophospholipids (PI) where it reached a plateau after 10 min incubation, while this incorporation was very weak in choline glycerophospholipids (PC). In contrast, the esterification of docosahexaenoic acid, another polyunsaturated fatty acid occurring at high level in brain, was similar in PI and PC, the incorporation in PI being 8-fold lower than that of 20:4n-6. The newly esterified [ 3H]20:4n-6 was exclusively found in the 1,2-diacyl subclasses of PI and PC. The bulk of incorporation was in the 18:0 20:4 n-6 molecular species of 1,2-diacyl-glycerophoinositol and in 16:0 20:4 n-6 + 18:1 20:4 n-6 molecular species of 1,2-diacyl-glycerophosphocholine, which agrees with the usual location of 20:4n-6 in brain phospholipid classes. Upon norepinephrine treatment, [ 3H]20:4n-6 was not released from PC, but was dose-dependently decreased in PI, the release being significant from 10 −5 M of the agonist. These results suggest that 20:4n-6 exhibits a high specific turnover in brain PI and is mobilized from this class upon relevant neuromediator stimulation. The acellular system used preserved the specificity of enzymes catalyzing the polyunsaturated fatty acid incorporation and release and could be helpful for studying their turn over in brain.