Identification of biological ligands of uranium in Danio rerio gonads : Impact on their function
Thesis: Uranium (U) is naturally presents at trace level (µg.L-1) in aquatic environment; its concentration can increase up to a few mg.L-1 due to human activities. Several ecotoxicological studies have shown uranium toxicity in contaminated zebrafishes Danio rerio, e.g. oxidative stress, genotoxicity and reprotoxicity (i.e. lower number of spawn and eggs laid) but mechanisms are not well known.The objective of this study is to contribute to the understanding of uranium reprotoxicity by elucidating the disrupted molecular mechanisms after contamination. Therefore, investigations have been carried out on ovaries from reproduced (R) and non-reproduced (NR) zebrafishes after waterborne exposure in laboratory conditions at environmentally relevant concentrations.This project was divided into two parts. Firstly, analytical investigations were carried out to continue the development of non-denaturing methods for U-protein identification by coupling separative techniques (size exclusion chromatography SEC, off gel electrophoresis OGE) with elemental (ICP MS) and molecular (ESI MS) sensitive mass spectrometry detection. Secondly, studies of U reprotoxicity were investigated by studying i) native U-protein complexes (metallomics approach) and ii) differential analysis of protein expression (proteomics approach)Analytical developments allowed keeping the physiological and non-denaturing extraction buffer for OGE separation step, improving U recovery. In ecotoxicology, the major results showed that ovary is an U accumulating organ and that the reproduction status modifies the accumulation level (R
Author(s)
Yvan Eb-Levadoux
Date of publication
- 2017
Keywords
- Ecotoxicology
- Uranium
- Reprotoxicity
- Zebrafish Danio rerio
- Uranium-protein complex
- Metallomics
- Elemental and molecular mass spectrometry coupling
- Non-denaturing analysis
- Proteomics
- Vitellogenin
- Oxidative stress
- Off gel electrophoresis
- SEC-ICP SFMS
- LC-ESI FTMS
Issuing body(s)
- Université de Pau et des Pays de l’Adour
Date of defense
- 03/04/2017
Thesis director(s)
- Ryszard Łobiński
URL of the HAL notice
Version
- 1