Highly Sensitive Quantitative Determination of Retinoic Acid Levels, Retinoic Acid Synthesis, and Catabolism in Embryonic Tissue Using a Reporter Cell-Based Method

Autor: Alisa S.W. Shum, Selina T K Tam, Peter McCaffery, Leo M Y Lee
Rok vydání: 2019
Předmět:
Zdroj: Methods in Molecular Biology ISBN: 9781493995844
DOI: 10.1007/978-1-4939-9585-1_13
Popis: 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.
Databáze: OpenAIRE