Retinoid metabolism in cultured human retinal pigment epithelium.
Uptake, esterification and release of all-trans-retinol in primary cultures of human retinal epithelium were studied. Cultured cells were supplemented with 3H-labelled 11,12-all-trans-retinol, using fatty-acid-free albumin as the carrier. This led to incorporation of retinal and the formation of all-trans- and 11-cis-retinyl palmitate. The metabolism of the all-trans ester was monitored in a medium containing various concentrations of foetal-bovine serum (FBS). In 20% (v/v) FBS, the ester was hydrolysed, and all-trans-retinol was released into the culture medium. In the absence of FBS, little ester was hydrolysed and no retinol was found in the medium. Dialysed or heat-inactivated FBS or fatty-acid-free albumin was as effective as FBS in provoking ester hydrolysis and retinol release. The concentration-dependency of this effect on FBS was matched by the corresponding concentrations of albumin alone. A linear relationship was also found between interphotoreceptor retinoid-binding protein and retinoid release. Haemoglobin, which does not bind retinoids, is ineffective in this capacity. It is concluded that lipid-binding substances, mainly albumin, in FBS act as acceptors for retinol and drain the cultured cells of this molecule. The release of the retinol is coupled to the hydrolysis of retinyl esters in the cell, so that there is little or no net hydrolysis of ester if there is no acceptor for retinol in the culture medium. This effect explains why cultured human retinal epithelial cells are depleted of their stores of retinoids when maintained in medium supplemented with FBS.
Das
S R
SR
Department of Ophthalmology, Columbia University, NY 10032.
Gouras
P
P
D001769:Blood; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004224:Diterpenes; D005136:Eye Proteins; D006801:Humans; D006868:Hydrolysis; D010857:Pigment Epithelium of Eye / Q000187:drug effects / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000494:pharmacology; D000084562:Retinyl Esters; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D014801:Vitamin A / Q000031:analogs & derivatives / Q000378:metabolism*; D000099214:Retinol-Binding Proteins, Interstitial
D004224:Diterpenes; D005136:Eye Proteins; D012177:Retinol-Binding Proteins; D000084562:Retinyl Esters; D012710:Serum Albumin, Bovine; D014801:Vitamin A; D000099214:Retinol-Binding Proteins, Interstitial; C014794:retinol palmitate