This study examined the effects of n-3 and n-6 polyunsaturated fatty acid alimentation on murine peritoneal macrophage phospholipids. Mice were fed complete diets supplemented with either corn oil predominantly containing 18:2n-6, borage oil containing 18:2n-6 and 18:3n-6, fish/corn oil mixture containing 18:2n-6, 20:5n-3 and 22:6n-3, or fish/borage oil mixture containing 18:2n-6, 18:3n-6, 20:5n-3 and 22:6n-3. After two weeks, the fatty acid levels of glycerophosphoserines (GPS), glycerophosphoinositols (GPI), sphingomyelin (SPH), and of the glycerophosphocholine (GPC) and glycerophosphoethanolamine (GPE) phospholipid subclasses were determined. We found that mouse peritoneal macrophage GPC contain primarily 1-O-alkyl-2-acyl (range for the dietary groups, 24.6-30.5 mol %) and 1,2-diacyl (63.2-67.2 mol %), and that GPE contains 1-O-alk-1'-enyl-2-acyl (40.9-47.4 mol %) and 1,2-diacyl (44.2-51.2 mol %) subclasses. In general, fish oil feeding increased macrophage 20:5n-3, 22:5n-3 and 22:6n-3 levels while simultaneously reducing 20:4n-6 in GPS, GPI, GPE and GPC subclasses except for 1-O-alk-1'-enyl-2-acyl GPC. Administration of 18:3n-6 rich diets (borage and fish/borage mixture) resulted in the accumulation of 20:3n-6 (2-carbon elongation product of 18:3n-6) in most phospholipids. In general, the novel combination of dietary 18:3n-6 and n-3 PUFA produced the highest 20:3n-6/20:4n-6 phospholipid fatty acid ratios. This study demonstrates that marked differences in the responses of macrophage phospholipid classes and subclasses exist following dietary manipulation.(ABSTRACT TRUNCATED AT 250 WORDS)