PUBLICATION

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

Authors
Eum, J., Kwak, J., Kim, H.J., Ki, S., Lee, K., Raslan, A.A., Park, O.K., Chowdhury, M.A., Her, S., Kee, Y., Kwon, S.H., Hwang, B.J.
ID
ZDB-PUB-161122-6
Date
2016
Source
International Journal of Molecular Sciences   17(11): (Journal)
Registered Authors
Chowdhury, Ashraf U., Eum, Juneyong, Kee, Yun, Kwak, Jina, Raslan, Ahmed A.
Keywords
3D live imaging, cardiac defect, fetal defect, light-sheet microscopy, pink water, single plane illumination microscopy, trinitrotoluene, zebrafish embryos
MeSH Terms
  • Animals
  • Apoptosis/drug effects
  • Embryo, Nonmammalian/drug effects
  • Embryo, Nonmammalian/pathology
  • Embryonic Development/drug effects*
  • Explosive Agents/toxicity*
  • Gene Expression/drug effects
  • Heart/drug effects
  • Heart/embryology
  • Heart Defects, Congenital/chemically induced
  • Heart Defects, Congenital/pathology
  • Intravital Microscopy
  • Melanocytes/drug effects
  • Trinitrotoluene/toxicity*
  • Water Pollutants, Chemical/toxicity*
  • Zebrafish
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed
27869673 Full text @ Int. J. Mol. Sci.
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.
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