Rapid quantification of neutral lipids and triglycerides during zebrafish embryogenesis.

P. Yoganantharjah, Avinesh Byreddy, Daniel Fraher, Munish Puri, Yann Gibert

    Research output: Contribution to journalArticle

    8 Citations (Scopus)

    Abstract

    The zebrafish is a useful vertebrate model to study lipid metabolism. Oil Red-O (ORO) staining of zebrafish embryos, though sufficient for visualizing the localization of triglycerides, was previously inadequate to quantify neutral lipid abundance. For metabolic studies, it is crucial to be able to quantify lipids during embryogenesis. Currently no cost effective, rapid and reliable method exists to quantify the deposition of neutral lipids and triglycerides.Thin layer chromatography (TLC), gas chromatography and mass spectrometry can be used to accurately measure lipid levels, but are time consuming and costly in their use. Hence, we developed a rapid and reliable method to quantify neutral lipids and triglycerides. Zebrafish embryos were exposed to Rimonabant (Rimo) or WIN 55,212-2 mesylate (WIN), compounds previously shown to modify lipid content during zebrafish embryogenesis. Following this, ORO stain was extracted out of both the zebrafish body and yolk sac and optical density was measured to give an indication of neutral lipid and triglyceride accumulation. Embryos treated with 0.3 µM WIN resulted in increased lipid accumulation, whereas 3 µM Rimo caused a decrease in lipid accumulation during embryogenesis. TLC was performed on zebrafish bodies to validate the developed method. In addition, BODIPY free fatty acids were injected into zebrafish embryos to confirm quantification of changes in lipid content in the embryo. Previously, ORO was limited to qualitative assessment; now ORO can be used as a quantitative tool to directly determine changes in the levels of neutral lipids and triglycerides.

    Original languageEnglish
    Pages (from-to)105-111
    Number of pages7
    JournalINTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY
    Volume61
    Issue number1-2
    DOIs
    Publication statusPublished - 2017

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