Effects of pregnancy and lactation on lipid metabolism in mouse mammary fat pads and nonmammary adipose tissues have been studied. In order to address the question whether the influence of hormonal milieu on lipid metabolism in mammary epithelial cells during pregnancy and lactation is the same as in fat cells, we have studied the mobilization of lipids and metabolism of fatty acids in the intact mammary glands, parenchyma-free mammary fat pads and in the perimetrial fat tissues of virgin, pregnant and lactating mice. Compared to parenchyma-free mammary fat pads, the perimetrial adipose tissues accumulated 5-fold higher levels of triglycerides during pregnancy. Mammary fat cells maintained overall lipid levels during pregnancy and lactation (16-20 micrograms/fat pad). In contrast, lactation depleted total lipid stores from 108 +/- 5 to 24 +/- 4.5 micrograms/fat pad in perimetrial fat pads. Results of comparative analysis of fatty acid composition of mammary fat pads, with and without epithelial tissue, from virgin and lactating mice showed stimulation of 18:2 omega 6 metabolism leading to 130% increase in the ratio 20:4 omega 6 to 18:2 omega 6 in the epithelial compartment. Pregnancy and lactation resulted in the elevation of 20:4 omega 6 levels probably due to a 4-fold increase in delta 5 desaturase activity and a decrease in oxidative degradation of 18:2 omega 6. These results suggest that, unlike other adipose tissues, the metabolic pathways in mammary fat cells are not dedicated to sequestration and accumulation of dietary lipids during pregnancy. Lactation favors mammary epithelial cell-stimulated production of precursors of eicosanoids which are known to have agonist-like effect on mammary epithelial cells.