Herein, studies concerning the biosynthesis, intracellular transport and utilization of isoprenoid lipids in vertebrate retinas are reviewed, with particular regard to rod photoreceptor cells and the assembly of rod outer segment (ROS) disk membranes. Initial in vitro studies with bovine retinas showed that [³H]mevalonate is metabolized primarily to squalene and ‘methylated’ sterols, rather than to cholesterol. Subsequently, similar results were obtained with frog retinas using [³H]acetate as a precursor, and the absolute rate of the sterol pathway was determined in vitro with ³H₂O. With the aid of vesicular transport inhibitors, energy poisons, and reduced temperature, it was demonstrated that lipid and protein trafficking mechanisms in the rod cell are separate and independent from one another. In vivo, the majority of newly synthesized squalene in the frog retina is not metabolized to sterols; rather, it is transported to the ROS, where it turns over in parallel with the disk membranes. The remaining squalene is converted slowly to cholesterol, much of which becomes incorporated into the ROS. In contrast, the in vivo metabolism of [³H]acetate to cholesterol in the rat retina is relatively efficient and rapid. However, in both frog and rat, retinal cholesterol turnover is slow (> 60 days), suggesting the existence of a retention mechanism that minimizes the need for de novo biosynthesis. The use of pharmacological approaches to assess the biological roles of isoprenoid lipids and protein prenylation in the retina and the mechanism of retinal cholesterol homeostasis are discussed.