Metabolomic Fingerprint Assay in Zebrafish Embryos

Abstract

In order for new drugs to enter the market, extensive studies are needed to examine toxic effects. Among others, teratogenicity studies are of paramount importance. Of even higher importance is to gain knowledge on the biological responses that take place upon drug exposure, so as to have a better understanding of the molecular mechanisms that govern developmental changes. Metabolomics is the research field that studies the changes in the chemical composition of metabolites contained within cells. Conducting metabolomics studies results in valuable information. Zebrafish is a vertebrate model organism that bridges in vivo assays and in vivo studies. In this chapter, we propose a metabolomic fingerprint assay for the study of metabolic changes in zebrafish embryos upon exposure to various drugs. The metabolome of zebrafish is extracted, and the ¹H-NMR spectrum is recorded. Using open-access metabolomic databases, a list of tentative metabolites is retrieved. The presence of the tentative metabolites is further confirmed by UHPLC-HRMS. Ultimately, after a metabolic pathway analysis, the metabolic network is revealed and useful conclusions can be drawn.

Keywords: 1H-NMR; Embryos; Fingerprint; Metabolite extraction; Metabolomics; Open-access databases; UHPLC-HRMS; Zebrafish.