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Analytical developments for the speciation of uranium in the zebrafish (Danio rerio) gills after exposure

Guillaume Bucher has defended his thesis on 22th November 2013 in Pau.

Document type > *Mémoire/HDR/Thesis

Keywords >

Research Unit > IRSN/PRP-ENV/SERIS/L2BT

Authors > BUCHER Guillaume

Publication Date > 22/11/2013

Summary

The objective of this thesis is to study the cellular compartmentalization and the chelation of uranium (U) by cytosolic proteins of gill cells of the zebrafish (Danio rerio, model specie in aquatic toxicology) under different direct exposure conditions (chronic vs. acute, 20 and 250 μg.L-1). This study required the development of hyphenated techniques (SEC, IEF off-gel, RP-UHPLC for the separation, ICP-SFMS, ESI-FTMS/MS for the detection) with the main challenges of maintaining the non-covalent U-biomolecule interactions and enhancing sensitivity for the analysis of environmentally relevant samples. After extraction, 24% to 32% of the total U detected in the gills were present in the cytosolic fraction, in which the U distribution on the biomolecules (as a function of their MW and pI) varied depending on the exposure level. Finally, U target biomolecules mapping allowed us (i) to highlight a particular affinity of U for acidic and/or P-containing proteins and (ii) to identify 24 protein candidates for U binding.

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Involved IRSN laboratory

Biochemistry, bioavailability and radionuclide transfer laboratory (L2BT)

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