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Int. J. Mol. Sci. 2016, 17(11), 1925; doi:10.3390/ijms17111925

3D Visualization of Developmental Toxicity of 2,4,6-Trinitrotoluene in Zebrafish Embryogenesis Using Light-Sheet Microscopy

1
Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea
2
Division of Environmental Research Center, Kangwon National University, Chuncheon 24341, Korea
3
Korea Basic Science Institute, Chuncheon Center, Chuncheon 24341, Korea
4
Department of Molecular Bioscience, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Academic Editors: Guido R.M.M. Haenen and Juliette Legler
Received: 31 August 2016 / Revised: 31 October 2016 / Accepted: 11 November 2016 / Published: 17 November 2016
(This article belongs to the Special Issue Zebrafish: A Model for Toxicological Research)
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Abstract

Environmental contamination by trinitrotoluene is of global concern due to its widespread use in military ordnance and commercial explosives. Despite known long-term persistence in groundwater and soil, the toxicological profile of trinitrotoluene and other explosive wastes have not been systematically measured using in vivo biological assays. Zebrafish embryos are ideal model vertebrates for high-throughput toxicity screening and live in vivo imaging due to their small size and transparency during embryogenesis. Here, we used Single Plane Illumination Microscopy (SPIM)/light sheet microscopy to assess the developmental toxicity of explosive-contaminated water in zebrafish embryos and report 2,4,6-trinitrotoluene-associated developmental abnormalities, including defects in heart formation and circulation, in 3D. Levels of apoptotic cell death were higher in the actively developing tissues of trinitrotoluene-treated embryos than controls. Live 3D imaging of heart tube development at cellular resolution by light-sheet microscopy revealed trinitrotoluene-associated cardiac toxicity, including hypoplastic heart chamber formation and cardiac looping defects, while the real time PCR (polymerase chain reaction) quantitatively measured the molecular changes in the heart and blood development supporting the developmental defects at the molecular level. Identification of cellular toxicity in zebrafish using the state-of-the-art 3D imaging system could form the basis of a sensitive biosensor for environmental contaminants and be further valued by combining it with molecular analysis. View Full-Text
Keywords: zebrafish embryos; pink water; trinitrotoluene; single plane illumination microscopy; light-sheet microscopy; 3D live imaging; cardiac defect; fetal defect zebrafish embryos; pink water; trinitrotoluene; single plane illumination microscopy; light-sheet microscopy; 3D live imaging; cardiac defect; fetal defect
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Eum, J.; Kwak, J.; Kim, H.J.; Ki, S.; Lee, K.; Raslan, A.A.; Park, O.K.; Chowdhury, M.A.U.; Her, S.; Kee, Y.; Kwon, S.-H.; Hwang, B.J. 3D Visualization of Developmental Toxicity of 2,4,6-Trinitrotoluene in Zebrafish Embryogenesis Using Light-Sheet Microscopy. Int. J. Mol. Sci. 2016, 17, 1925.

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