(13)C-isotope-based protocol for prenyl lipid metabolic analysis in zebrafish embryos
- Authors
- Mugoni, V., Medana, C., and Santoro, M.M.
- ID
- ZDB-PUB-131122-19
- 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.
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.