ZFIN ID: ZDB-PUB-131122-19
(13)C-isotope-based protocol for prenyl lipid metabolic analysis in zebrafish embryos
Mugoni, V., Medana, C., and Santoro, M.M.
Date: 2013
Source: Nature Protocols   8(12): 2337-2347 (Journal)
Registered Authors: Mugoni, Vera, Santoro, Massimo
Keywords: none
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
PubMed: 24177291 Full text @ Nat. Protoc.
ABSTRACT

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 13C-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.

ADDITIONAL INFORMATION