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Effects of Methylmercury on Harbour Seal Peripheral Blood Leucocytes In Vitro Studied by Electron Microscopy

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Abstract

Methylmercury (MeHg) is highly immunotoxic and can alter the health status of the harbour seal, Phoca vitulina, from the North Sea. To investigate the mechanism of MeHg-induced toxicity in harbour seal lymphocytes, Concanavalin A (ConA)-stimulated peripheral blood leucocytes were exposed in vitro to sublethal concentrations of MeHgCl (0.2, 1, and 2 µM) for 72 h and then analysed for their viability and ultrastructure. After 72 h of incubation, cells were counted with a propidium iodide staining technique, a metabolic MTS assay was performed, and cells exposed to 1 µM of MeHgCl were observed by transmission electron microscopy (TEM). Alive cell numbers decreased with increased MeHgCl concentrations. In presence of ConA and 1 µM of MeHgCl, TEM images revealed a higher frequency of apoptotic cells. Exposed cells displayed condensation of the chromatin at the nuclear membrane and mitochondrial damages. The results suggest that in vitro MeHgCl-induced apoptosis in harbour seal lymphocytes through a mitochondrial pathway.

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Acknowledgments

Krishna Das and Joseph Schnitzler are supported by F.R.S. – FNRS Research Associate (Fonds de la Recherche Scientifique). Aurélie Dupont was supported by a FRIA grant (Fonds pour la formation à la Recherche dans l’Industrie et dans l’Agriculture). The seal capture was conducted under the funding of the Ministry of Renewable Energies, Agriculture, Environment and Rural Areas of Schleswig–Holstein (MELUR) and Schleswig–Holstein’s Government-Owned Company for Coastal Protection, National Parks and Ocean Protection (LKN-SH), Germany. The authors thank Dr. Jörg Driver for blood analyses and interpretation. Thanks also to Tanja Rosenberger (Seal Centre Friedrichskoog, Germany) and all volunteers involved in seal sampling. The authors also are grateful to Philippe Compère and Nicole Decloux (Laboratoire de Morphologie Fonctionnelle et Evolutive, ULg) for their valuable assistance in electron microscopy analysis. The authors are grateful to the anonymous referees for their useful comments on an earlier draft of this manuscript. This is a MARE publication 300.

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Authors and Affiliations

  1. Laboratory of Oceanology – MARE Center, University of Liege, B6c, Liège, Belgium

    Aurélie Dupont, Joseph Schnitzler & Krishna Das

  2. Mammalian Cell Culture Laboratory (GIGA-R), University of Liege, B6c, Liège, Belgium

    Marie-Claire De Pauw-Gillet

  3. Institute for Terrestrial and Aquatic Wildlife Research, University of Veterinary Medicine Hannover, Foundation, Werftstrasse 6, 25761, Buesum, Germany

    Ursula Siebert

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  1. Aurélie Dupont
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  5. Krishna Das
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Correspondence to Krishna Das.

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Dupont, A., De Pauw-Gillet, MC., Schnitzler, J. et al. Effects of Methylmercury on Harbour Seal Peripheral Blood Leucocytes In Vitro Studied by Electron Microscopy. Arch Environ Contam Toxicol 70, 133–142 (2016). https://doi.org/10.1007/s00244-015-0207-y

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