Uptake, processing and release of retinoids by cultured human retinal pigment epithelium.
Upon absorption of a photon, the 11-cis retinaldehyde chromophore of rhodopsin is isomerized and reduced to all-trans retinol (vitamin A) in the photoreceptor outer segments, whereupon it leaves the photoreceptors, and moves to the retinal pigment epithelium (RPE). To clarify the function of the RPE in the regeneration of 11-cis retinaldehyde, we delivered all-trans retinol to monolayer cultures of human RPE. During delivery the retinol was associated with its putative natural carrier, interphotoreceptor retinoid binding protein (IRBP). IRBP has been proposed as a carrier protein involved in the exchange of retinoids between the photoreceptors and the retinal pigment epithelium. The retinoid composition of RPE cells and culture medium was analyzed by HPLC following several incubation periods. The RPE monolayer was found to process all-trans retinol into two distinct end-products: all-trans retinyl palmitate, which remained within the RPE monolayer: and 11-cis retinaldehyde which was released into the culture medium. These results demonstrate retinoid isomerase, retinol oxidoreductase and retinyl ester synthetase activity in human RPE cells cultured under the appropriate conditions. They show that IRBP can serve as a carrier of retinol through an aqueous medium to the RPE, and they illustrate that the visual cycle can be studied in vitro.
Flannery
J G
JG
Jules Stein Eye Institute UCLA School of Medicine 90024.
O'Day
W
W
Pfeffer
B A
BA
Horwitz
J
J
Bok
D
D
D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004224:Diterpenes; D005136:Eye Proteins / Q000378:metabolism; D006801:Humans; D010857:Pigment Epithelium of Eye / Q000737:chemistry / Q000378:metabolism*; D012172:Retinaldehyde / Q000378:metabolism; D012176:Retinoids / Q000032:analysis / Q000493:pharmacokinetics*; D012177:Retinol-Binding Proteins / Q000378:metabolism; D000084562:Retinyl Esters; D013997:Time Factors; D014801:Vitamin A / Q000031:analogs & derivatives / Q000378:metabolism / Q000493:pharmacokinetics; D000099214:Retinol-Binding Proteins, Interstitial
D004224:Diterpenes; D005136:Eye Proteins; D012172:Retinaldehyde; D012176:Retinoids; D012177:Retinol-Binding Proteins; D000084562:Retinyl Esters; D014801:Vitamin A; D000099214:Retinol-Binding Proteins, Interstitial; C014794:retinol palmitate