Brain docosahexaenoic acid (DHA, 22:6n–3) levels are associated with learning memory performance, but it is not known the mechanism of DHA on enhancing memory effect. The aim of this study was to examine effect of brain DHA levels on cytoskeleton expression. Rats were fed a chow or sunflower oil-based n–3 fatty acid- deficient diet supplemented with or without fish oil starting from embryo and through postnatal day 140. The various DHA levels were from 5.0% to 15.6% of total fatty acids in hippocampus, 3.9% to 13.7% in visual cortex, and 5.3% to 14.4% in olfactory bulbs. The expression of the cytoskeleton markers tyrosine tubulin, acetylated tu- bulin, and β-actin in the hippocampus, visual cortex and olfactory bulb was not affected by brain DHA levels.