13C-isotope-based protocol for prenyl lipid metabolic analysis in zebrafish embryos

Nat Protoc. 2013 Dec;8(12):2337-47. doi: 10.1038/nprot.2013.139. Epub 2013 Oct 31.


Metabolism has a decisive role in many fundamental biological processes, including organism development and tissue homeostasis. Here we describe a protocol for fast and reliable (13)C-isotope-based in vivo metabolic profiling. This protocol covers the loading of isotope precursor; extraction, preparation and quantification of the labeled lipid metabolites (e.g., the prenyl lipid CoQ10) by the means of HPLC-MS; and its analysis in zebrafish embryos. This protocol can be applied to different types of experimental settings, including tissue-specific metabolic analyses or dynamic metabolic changes that occur during vertebrate embryogenesis. The protocol takes 5-7 d to complete, requiring minimal equipment and analytical expertise, and it represents a unique alternative to the existing ex vivo (e.g., cell lines) isotope-based metabolic methods. This procedure represents a valuable approach for researchers interested in studying the effect of gene manipulation on lipid metabolism in zebrafish and in understanding the genetic conditions that result in metabolism dysfunction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbon Isotopes
  • Chromatography, High Pressure Liquid
  • Dimethylallyltranstransferase / genetics
  • Dimethylallyltranstransferase / metabolism
  • Lipid Metabolism*
  • Mass Spectrometry
  • Metabolome*
  • Metabolomics / methods*
  • Prenylation
  • Ubiquinone / analogs & derivatives
  • Ubiquinone / chemistry
  • Ubiquinone / metabolism
  • Zebrafish / genetics
  • Zebrafish / metabolism*
  • Zebrafish Proteins / genetics
  • Zebrafish Proteins / metabolism


  • Carbon Isotopes
  • Zebrafish Proteins
  • Ubiquinone
  • Dimethylallyltranstransferase
  • Ubiad1 protein, zebrafish
  • coenzyme Q10