PUBLICATION

In vivo identification of potential uranium protein targets in zebrafish ovaries after chronic waterborne exposure

Authors
Eb-Levadoux, Y., Frelon, S., Simon, O., Arnaudguilhem, C., Lobinski, R., Mounicou, S.
ID
ZDB-PUB-170321-8
Date
2017
Source
Metallomics : integrated biometal science   9(5): 525-534 (Journal)
Registered Authors
Keywords
none
MeSH Terms
  • Animals
  • Chromatography, Gel
  • Female
  • Fish Proteins/analysis*
  • Fish Proteins/metabolism
  • Metalloproteins/analysis*
  • Metalloproteins/metabolism
  • Models, Molecular
  • Ovary/drug effects
  • Ovary/metabolism*
  • Proteomics/methods
  • Spectrometry, Mass, Electrospray Ionization
  • Uranium/analysis*
  • Uranium/metabolism
  • Uranium/toxicity
  • Water Pollutants, Chemical/analysis*
  • Water Pollutants, Chemical/metabolism
  • Water Pollutants, Chemical/toxicity
  • Zebrafish
PubMed
28317950 Full text @ Metallomics
Abstract
Ecotoxicological studies have indicated the reprotoxicity of uranium (U) in zebrafish, but its molecular mechanisms remain unclear. Due to the non-covalent nature of U-protein complexes, canonical proteomics approaches are often not relevant as they usually use denaturating reagents or solvents. In this study, non-denaturating (ND) methods were used to obtain insight into the nature of U potential targets in ovaries of reproduced and non-reproduced zebrafish after 20 days of exposure to an environmentally relevant U concentration (20 μg L-1). After the ND sample preparation, 1-dimensional (SEC) and 2-dimensional (OGE × SEC) separations followed by ICP-sector-field MS measurements (U, P, Fe, Cu, and Zn) enabled the determination of chemical characteristics (MW, pI) of the metal-protein complexes. Phosphorus and U coelution confirmed the affinity of U for P-containing proteins. In addition, 2D separation allowed the discrimination of Fe-metalloproteins as potential U targets. Finally, 20 protein candidates for U complexation were identified after tryptic digestion conditions by LC-ESI FT MS and a database search. Potential U targets were mainly involved in three biological processes: oxidative stress regulation (SOD, GST), cytoskeleton structure (actin) and embryo early development (vtg, initiation factor).
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping