Retinol Research Today is a free monthly online journal that collates and summarizes the latest research about Retinol, including details on vitamin a, uses, wrinkle treatment, anti-aging.

Development of a versatile reporter assay for studies of retinol uptake and metabolism in vivo.

Lidén M, Eriksson U

Ludwig Institute for Cancer Research, Stockholm Branch, Box 240, S-171 77 Stockholm, Sweden.

The two isomers of retinoic acid (RA), all-trans RA and 9-cis RA, are produced in several tissues in order to allow specific control of target gene transcription. Given the high potency of these receptor ligands, it seems likely that the cellular uptake and metabolic activation of the precursor, retinol (vitamin A), should be a highly regulated process. Several retinol dehydrogenases and components involved in the downstream events have been identified and partially characterized. However, less is known about the cellular uptake of retinol, and the isomerase activity giving rise to the 9-cis and 11-cis branches of the pathway. In this work, we show that the 9-cis RA biosynthesis pathway can be fully reconstituted in cultured HEK293A cells expressing a reporter system, including an endogenous isomerase activity converting all-trans retinol into 9-cis retinol. This assay allows for functional studies of known components, as well as screening for yet unidentified genes involved in the pathway. In addition to free all-trans retinol, we find that these cells can take up retinol from plasma retinol binding protein (RBP) by a mechanism that can be efficiently inhibited by blocking antibodies, suggesting that the uptake may involve a cellular receptor. We also demonstrate that overexpression of CRBPI can drive the accumulation of intracellular retinol from unbound retinol added to the medium. Thus, this versatile cellular assay can be used to study several aspects of retinol uptake and metabolism in vivo.

Published 21 October 2005 in Exp Cell Res, 310(2): 401-8.
Full-text of this article is available online (may require subscription).

© 2004-2008 Retinol Research Today. All Rights Reserved.