In this study, we examined the effects of dietary fatty acids on the fatty acid composition of phospholipid fractions in regions of the brain and on behavioral development in rats. The Sprague Dawley rats were fed the experimental diets 3~4 wks prior to the conception. Experimental diets consisted of 10% fat(wt/wt) which were from either safflower oil (SO, poor in ω3 fatty acids), mixed oil (MO, P/M/S ratio = 1:1.4:1, ω6/ω3 ratio = 6.3), or mixed oil supplemented with vitamin E (+500 mg/kg diet). At 3 and 9 weeks of age, frontal cortex (FC), corpus striatum (CS), hippocampus (H), and cerebellum (CB) were dissected from the whole brain. The fatty acid content was determined in the different phospholipid fractions: phosphatidylcholine (PC), phosphatidylserine (PS), and phosphatidylethanolamine (PE) in the rat brain regions. In the visual discrimination test, the order of the cumulative errors made in Y-water maze test were SO>MO>ME. This suggested that the balanced diet supplemented with vitamin E had the most beneficial effect on learning ability. The overall characteristics of correlation between fatty acids and behavior development were that the frequency of cumulative errors were negatively correlated significantly with monounsaturated fatty acids (MUFAs), ie., 18:1 ω9 and 22:1 ω9. Docosahexaenoic acid (22:6ω3) of PS in frontal cortex (FC) was negatively correlated with the number of errors made in the Y-water maze test. 22:5ω6 PS in hippocampus (H), PC and PE in corpus striatum (CS), PC in cerebellum (CB) were positively correlated with cumulative errors. And these errors were negatively correlated with 20:4ω6 of PE in corpus striatum (CS) and PC in cerebellum (CB). Especially, Oleic acid (18:1ω9) in all phospholipid fractions (PC, PS, PE) of hippocampus was negatively correlated with the number of errors. These findings demonstrate that the MUFAs were might be essential for proper brain development, especially in hippocampus which is generally thought to be the regions of memory and learning.