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

Leo M.Y. Lee, Selina T.K. Tam, Peter J. McCaffery, Alisa S.W. Shum

Research output: Chapter in book / Conference proceedingChapter in an edited book (as author)Academic researchpeer-review

2 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages181-192
Number of pages12
DOIs
Publication statusPublished - 2019
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2019
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cell culture
  • CYP26
  • Embryo
  • RALDH
  • Retinoic acid
  • Retinoic acid metabolism
  • Retinoic acid reporter cell line
  • Retinoic acid synthesis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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