| Summary: | Since the beginning of the 1980s, stocks of European eel have been declining in most of their geographical distribution area. Many factors can be attributed to this decline such as pollution by xenobiotics released into the environment through agricultural, industrial and domestic activities. Because the New European Chemicals Legislation (REACh) is asking for alternatives to animal testing and reduction of animals sacrified in ecotoxicology and in accordance with conservation biology considerations, we have developed an appropriate and reproducible methodology to obtain a post-nuclear fraction of isolated European eel peripheral blood mononuclear cells (PBMC) in order to evaluate the toxicity of xenobiotics using a subproteomic approach. In a first study, we have studied the in vitro toxicity of perfluorooctane sulfonate (PFOS) in eel PBMC exposed during 48 h to sublethal concentrations (10 µg and 1 mg PFOS/L). A proteomic analysis using 2D-DIGE was performed to compare PBMC from the control group with cells exposed to the pollutant. A total of 48 proteins displaying significant changes in abundance were identified and categorized according to their functional classes. Besides providing clues on the cellular pathways mainly affected by PFOS, results allowed the identification of proteins rarely found in other ecotoxicological proteomic studies. These proteins could constitute potential biomarkers of exposure to PFOS in fish. In order to determine the specificity of the proteomic pattern observed after in vitro PFOS contaminations, we have completed the set of data with in vitro exposures to two other xenobiotics, dichlorodiphenyl-trichloroethane (DDT) and cadmium, using exactly the same methodologies as for the PFOS experiments. The aim of this new study was the discovery of protein expression signatures specific of different classes of pollutants. The identification of the proteins of interest by mass spectrometry allowed selecting four candidates for a minimal common signature between the three experiments. ...
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