Ocular tissues are highly dependent on lipid turnover and metabolism, which requires an uptake mechanism for fatty acids from lipoproteins. We studied the activity and expression of lipoprotein lipase (LPL), which catalyzes the hydrolysis of plasma triglycerides, in different ocular regions. Human and bovine eyes were dissected and various specialized anatomical areas were assayed for LPL activity, mRNA, and immunoreactivity. Variable levels of LPL activity were detected in all structures in human and bovine eyes. LPL activity was much higher in vascularized structures, such as ciliary body, iris, and retina than in avascular eye structures, such as vitreous body, lens, and cornea. In both human and bovine eyes, ciliary body contained the highest LPL lipolytic activity. LPL mRNA was detected by reverse transcription followed by polymerase chain reaction (RT-PCR) in all regions of human eyes. By RT-PCR analysis it was shown that bovine eyes contained high levels of LPL mRNA in ciliary body and iris, lower levels were found in retina, optic nerve, and lens, whereas no LPL mRNA could be found in bovine cornea. RT-PCR data, obtained in bovine eyes, agree with the results obtained by Northern blot experiments, confirming the high levels of LPL mRNA in iris and ciliary body. Immunofluorescence experiments performed on human eye samples indicated that the LPL protein is mostly distributed on the choroides, the choriocapillaris, and on the vessels of ciliary body, iris, optic nerve, and retina. The present study demonstrates that active LPL protein is synthesized, secreted, and located among microvessels in several specialized regions of the eye, and suggests that LPL could be involved in the uptake of fatty acids by the ocular tissues.