Use of fish otoliths as a temporal biomarker of field uranium exposure.

This study aimed to determine uranium (U) pollution over time using otoliths as a marker of fish U contamination. Experiments were performed in field contamination (~20 µg L-1: encaged fish: 15d, 50d and collected wild fish) and in laboratory exposure conditions (20 and 250 µg L-1, 20d). We reported the U seasonal concentrations in field waterborne exposed roach fish (Rutilus rutilus), in organs and otoliths. Otoliths were analyzed by ICPMS and LA-ICP SF MS of the entire growth zone. Concentrations were measured on transects from nucleus to the edge of otoliths to characterize environmental variations of metal accumulation. Results showed a spatial and temporal variation of U contamination in water (from 51 to 9.4 µg L-1 at the surface of the water column), a high and seasonal accumulation in fish organs, mainly the digestive tract (from 1000 to 30,000 ng g-1, fw), the gills (from 1600 to 3200 ng g-1, fw) and the muscle (from 144 to 1054 ng g-1, fw). U was detected throughout the otolith and accumulation varied over the season from 70 to 350 ng g-1, close to the values measured (310 ng g-1) after high exposure levels in laboratory conditions. U in otoliths of encaged fish showed rapid and high U accumulation from 20 to 150 ng g-1. The U accumulation signal was mainly detected on the edge of the otolith, showing two U accumulation peaks, probably correlated to fish age, i.e. 2 years old. Surprisingly, elemental U and Zn signatures followed the same pattern therefore using the same uptake pathways. Laboratory, caging and field experiments indicated that otoliths were able to quickly accumulate U on the surface even for low levels and to store high levels of U. This study is an encouraging first step in using otoliths as a marker of U exposure.

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

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).