title: 3D Visualization of Developmental Toxicity of 2,4,6-Trinitrotoluene in Zebrafish Embryogenesis Using Light-Sheet Microscopy.
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.
authors.affiliation: Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea. eum1230@naver.com.
authors.forename: Juneyong
authors.initials: J
authors.lastname: Eum
authors.affiliation: Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea. wlswns02@naver.com.
authors.forename: Jina
authors.initials: J
authors.lastname: Kwak
authors.affiliation: Division of Environmental Research Center, Kangwon National University, Chuncheon 24341, Korea. polarbear723@naver.com.
authors.forename: Hee Joung
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea. tjdud729@naver.com.
authors.forename: Seoyoung
authors.initials: S
authors.lastname: Ki
authors.affiliation: Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea. lky@kangwon.ac.kr.
authors.forename: Kooyeon
authors.initials: K
authors.lastname: Lee
authors.affiliation: Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea. ahmed.raslan8080@gmail.com.
authors.forename: Ahmed A
authors.initials: AA
authors.lastname: Raslan
authors.affiliation: Korea Basic Science Institute, Chuncheon Center, Chuncheon 24341, Korea. okpark81@kbsi.re.kr.
authors.forename: Ok Kyu
authors.initials: OK
authors.lastname: Park
authors.affiliation: Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea. ashraf.rx@gmail.com.
authors.forename: Md Ashraf Uddin
authors.initials: MA
authors.lastname: Chowdhury
authors.affiliation: Korea Basic Science Institute, Chuncheon Center, Chuncheon 24341, Korea. swher@kbsi.re.kr.
authors.forename: Song
authors.initials: S
authors.lastname: Her
authors.affiliation: Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea. yunkee@kangwon.ac.kr.
authors.forename: Yun
authors.initials: Y
authors.lastname: Kee
authors.affiliation: Korea Basic Science Institute, Chuncheon Center, Chuncheon 24341, Korea. kwonsh@kbsi.re.kr.
authors.forename: Seung-Hae
authors.initials: SH
authors.lastname: Kwon
authors.affiliation: Department of Molecular Bioscience, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea. bjhwang@kangwon.ac.kr.
authors.forename: Byung Joon
authors.initials: BJ
authors.lastname: Hwang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000473:pathology; D047108:Embryonic Development / Q000187:drug effects*; D053834:Explosive Agents / Q000633:toxicity*; D015870:Gene Expression / Q000187:drug effects; D006321:Heart / Q000187:drug effects / Q000196:embryology; D006330:Heart Defects, Congenital / Q000139:chemically induced / Q000473:pathology; D000069416:Intravital Microscopy; D008544:Melanocytes / Q000187:drug effects; D014303:Trinitrotoluene / Q000633:toxicity*; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D053834:Explosive Agents; D014874:Water Pollutants, Chemical; D029961:Zebrafish Proteins; D014303:Trinitrotoluene
keywords: 3D live imaging; cardiac defect; fetal defect; light-sheet microscopy; pink water; single plane illumination microscopy; trinitrotoluene; zebrafish embryos