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Volume 271, Number 25, Issue of June 21, 1996 pp. 14971-14980
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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An Analysis of Retinoic Acid-induced Gene Expression and Metabolism in AB1 Embryonic Stem Cells

(Received for publication, January 30, 1996, and in revised form, March 28, 1996)

From the Department of Pharmacology, Cornell University Medical College, New York, New York 10021

Murine embryonic stem cells such as the AB1 cell line undergo differentiation in the presence of retinoic acid (RA) into an extraembryonic epithelial cell type. This results in the activation of genes such as Hoxa-1, Hoxb-1, laminin, collagen IV(1), tissue plasminogen activator, RAR, and CRABPII. The CRABPI gene is regulated in an unusual fashion; CRABPI message and protein levels are induced at low concentrations of RA, but induction is diminished at higher concentrations. AB1 cells take up RA rapidly from the medium, and the addition of low, exogenous concentrations of RA to the culture medium results in very high intracellular RA concentrations. For example, AB1 stem cells cultured in 5 nM [³H]RA have an internal [³H]RA concentration of 1-2 µM within the first hour. AB1 cells also metabolize [³H]RA to more polar RA derivatives. The half-life of RA in AB1 cells not previously exposed to RA is about 2-2.5 h versus 40-45 min in cells cultured for 2-3 days in 1 µM exogenous RA. Thus, the enzyme(s) which metabolize RA are induced or activated by RA. Furthermore, the local concentration of RA required to elicit some biological responses may be higher than previously thought.

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