Abstract
The effect of all-trans retinoic acid (RA) on embryogenesis is tissue specific and highly concentration dependent. Using a liquid chromatography/mass spectrometry-based method to quantify trace amounts of RA in embryonic tissue requires expensive specialist facilities. Here, we describe the use of a RA response element (RARE)-lacZ reporter cell-based method, which is simple and cost effective, to measure RA levels in small pieces of tissue from the embryo. We further apply this method to quantitatively assay activities of RA-synthesizing and RA-catabolizing enzymes, the key regulators of RA bioavailability in tissues and developing organs of the embryo.
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Acknowledgments
This work was supported by the Hong Kong Research Grants Council General Research Fund Ref No. 14113314 and 14174017 to A.S.W.S. and P.J.M. We thank Dr. Michael Wagner (SUNY Downstate Medical Center) for the gift of the F9 RARE-lacZ cell line, and Johnson & Johnson for the gift of R115866.
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Lee, L.M.Y., Tam, S.T.K., McCaffery, P.J., Shum, A.S.W. (2019). Highly Sensitive Quantitative Determination of Retinoic Acid Levels, Retinoic Acid Synthesis, and Catabolism in Embryonic Tissue Using a Reporter Cell-Based Method. In: Ray, S. (eds) Retinoid and Rexinoid Signaling . Methods in Molecular Biology, vol 2019. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9585-1_13
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DOI: https://doi.org/10.1007/978-1-4939-9585-1_13
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