Lipoprotein lipase (LP lipase, triacylglyceroprotein acyihydrolase EC 3.1.1.34) activity was found in four dissimilar brain regions (hypothalamus, cortex, cerebellum, and midbrain) of adult male rats. Progressive accumulation of LP lipase activity in cultured fetal rat hypothalamic cells was also observed, indicating de novo synthesis of the lipase. The brain LP lipase activity was serum-dependent and was inhibited by 1 (2) or assimilated from the plasma compartment either from albumin-bound nonesterified fatty acids or from fatty acids liberated by triacylglycerol-rich lipoprotein (chylomicron or very low density lipoprotein) triacylglycerol hydrolysis. This hydrolysis would be dependent on lipoprotein lipase (LP lipase, triacylglycero-protein acylhydrolase, EC 3.1.1.34), an enzyme produced in a number of extracranial tissues, including muscle, adipose tissue, and lactating breast (3). Smaller amounts of LP lipase have been detected in rat brain (4). However, in studies of rabbit brain microvessels, the enzyme was examined only on the endothelial surface (5), the site of triacylglycerol-rich lipoprotein triacylglycerol hydrolysis in other tissues (3).Brain LP lipase could not only be an important source of fatty acids for brain cell lipid but could also serve as a metabolic regulator. For instance, increases in LP lipase-mediated fatty acid delivery to the hypothalamus could be a mechanism by which alterations in food intake are regulated. We provide evidence that LP lipase is present in several brain regions, is synthesized by and functional in brain cells in vitro, and is subject to regulation in vivo by fasting. It is the latter observation that strongly suggests a regulatory role of the enzyme in the brain. Hypothalamic Cell Cultures. Hypothalamic cell cultures were prepared from the ventral diencephalons of 16-day-old rat embryos, using techniques previously reported for culturing rat telencephalic cells (7). The hypothalamic cells were prepared by enzymatic (neutral protease/papain/deoxyribonuclease) dispersal of a fragment of embryonic brain obtained as follows: fetal brains were placed on their dorsal surfaces and 1-mm-deep cuts were made along the lateral hypothalamic sulci, anterior to the optic chiasm, and caudal to the mammillary bodies. Choroid lining the third ventricle and surface meninges were removed prior to enzymatic dispersal. Polystyrene flasks (Coming, 25 cm2) were each seeded with 5 x 106 cells on day 0. Cultures were maintained in minimal essential medium (Eagle) with Earle's salts plus 10% fetal calf serum at 370C in a 5% CO2 atmosphere. After 96 hr, the fetal calf serum was replaced with heat-inactivated horse serum. (All media and sera were obtained from GIBCO.) These cultures are morphologically heterogeneous, containing neuronal, glial, meningeal, choroidal, and, possibly, vascular cells. The non-neuronal cells proliferate and cover the surface of the flask by 96 hr. Neuronal morphology is mature and stable between days 8 and 20 in vitro. Cells from one 8-day-old culture con...