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Developmental toxicity and oxidative stress induced by gamma irradiation in zebrafish embryos

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Abstract

This study aimed to evaluate the biological effects of gamma irradiation on zebrafish embryos. Different doses of gamma rays (0.01, 0.05, 0.1, 0.5 and 1 Gy) were used to irradiate zebrafish embryos at three developmental stages (stage 1, 6 h post-fertilization (hpf); stage 2, 12 hpf; stage three, 24 hpf), respectively. The survival, malformation and hatching rates of the zebrafish embryos were measured at the morphological endpoint of 96 hpf. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione S-transferase (GST) were assayed. Morphology analysis showed that gamma irradiation inhibited hatching and induced developmental toxicity in a dose-dependent manner. Interestingly, after irradiation the malformation rate changed not only in a dose-dependent manner but also in a developmental stage-dependent manner, indicating that the zebrafish embryos at stage 1 were more sensitive to gamma rays than those at other stages. Biochemical analysis showed that gamma irradiation modulated the activities of antioxidant enzymes in a dose-dependent manner. A linear relationship was found between GPx activity and irradiation dose in 0.1–1 Gy group, and GPx was a suitable biomarker for gamma irradiation in the dose range from 0.1 to 1 Gy. Furthermore, the activities of SOD, CAT, GR and GPx of the zebrafish embryos at stage 3 were found to be much higher than those at other stages, indicating that the zebrafish embryos at stage 3 had a greater ability to protect against gamma rays than those at other stages, and thus the activities of antioxidant enzymes changed in a developmental stage-dependent manner.

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Abbreviations

BCA:

Bicinchoninic acid

hpf:

Hours post-fertilization

XOD:

Xanthine oxidase

SOD:

Superoxide dismutase

GSSG:

L-Glutathione oxidized

CAT:

Catalase

GSH:

Reduced glutathione

GR:

Glutathione reductase

NADPH:

Nicotinamide adenine dinucleotide phosphate

GPx:

Glutathione peroxidase

DTNB:

5,5′-Dithiobis-2-nitrobenzoic acid

GST:

Glutathione S-transferase

CDNB:

1-Chloro-2,4-dinitrobenzene

ROS:

Reactive oxygen species

LPO:

Lipid hydroperoxide

References

  • Bai W, Zhang Z, Tian W, He X, Ma Y, Zhao Y, Chai Z (2010) Toxicity of zinc oxide nanoparticles to zebrafish embryo: a physicochemical study of toxicity mechanism. J Nanopart Res 12:1645–1654

    Article  Google Scholar 

  • Barbazuk WB, Korf I, Kadavi C, Heyen J, Tate S, Wun E, Bedell JA, McPherson JD, Johnson SL (2000) The syntenic relationship of the zebrafish and human genomes. Genome Res 10:1351–1358

    Article  Google Scholar 

  • Bladen CL, Flowers MA, Miyake K, Podolsky RH, Barrett JT, Kozlowski DJ, Dynan WS (2007) Quantification of ionizing radiation-induced cell death in situ in a vertebrate embryo. Radiat Res 168:149–157

    Article  Google Scholar 

  • Bourrachot S, Simon O, Gilbin R (2008) The effects of waterborne uranium on the hatching success, development, and survival of early life stages of zebrafish (Danio rerio). Aquat Toxicol 90:29–36

    Article  Google Scholar 

  • Celik I, Tuluce Y, Turker M (2006) Antioxidant and immune potential marker enzymes assessment in the various tissues of rats exposed to indoleacetic acid and kinetin: a drinking water study. Pestic Biochem Physiol 86:180–185

    Article  Google Scholar 

  • Choi VWY, Yu KN (2015) Embryos of the zebrafish Danio rerio in studies of non-targeted effects of ionizing radiation. Cancer Lett 356:91–104

    Article  Google Scholar 

  • Choi VWY, Konishi T, Oikawa M, Iso H, Cheng SH, Yu KN (2010) Adaptive response in zebrafish embryos induced using microbeam protons as priming dose and X-ray photons as challenging dose. J Radiat Res 51:657–664

    Article  Google Scholar 

  • Choi VWY, Yum EHW, Konishi T, Oikawa M, Cheng SH, Yu KN (2012) Triphasic low-dose response in zebrafish embryos irradiated by microbeam protons. J Radiat Res 53:475–481

    Google Scholar 

  • Choi VWY, Konishi T, Oikawa M, Cheng SH, Yu KN (2013a) The threshold number of protons to induce an adaptive response in zebrafish embryos. J Radiol Prot 33:91–100

    Article  Google Scholar 

  • Choi VWY, Ng CYP, Kobayashi A, Konishi T, Suya N, Ishikawa T, Cheng SH, Yu KN (2013b) Bystander effect between zebrafish embryos in vivo induced by high-dose X-rays. Environ Sci Technol 47:6368–6376

    Google Scholar 

  • Chung HC, Kim SH, Lee MG, Cho CK, Kim TH, Lee DH, Kim SG (2001) Mitochondrial dysfunction by gamma-irradiation accompanies the induction of cytochrome P450 2E1 (CYP2E1) in rat liver. Toxicology 161:79–91

    Article  Google Scholar 

  • Clemente Z, Castro VL, Moura MA, Jonsson CM, Fraceto LF (2014) Toxicity assessment of TiO2 nanoparticles in zebrafish embryos under different exposure conditions. Aquat Toxicol 147:129–139

    Article  Google Scholar 

  • Cunliffe VT (2003) Zebrafish: a Practical Approach. Genet Res 82:79–79

    Article  Google Scholar 

  • Egami N, Shimada A, Hama-Furukawa A (1983) Dominant lethal mutation rate after gamma-irradiation of the fish, Oryzias latipes. Mutat Res 107:265–277

    Article  Google Scholar 

  • Fan J, Shi M, Huang JZ, Xu J, Wang ZD, Guo DP (2014) Regulation of photosynthetic performance and antioxidant capacity by 60Co gamma-irradiation in Zizania latifolia plants. J Environ Radioact 129:33–42

    Article  Google Scholar 

  • Foyer CH, Halliwell B (1976) The presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. Planta 133:21–25

    Article  Google Scholar 

  • Garner LV, Di Giulio RT (2012) Glutathione transferase pi class 2 (GSTp2) protects against the cardiac deformities caused by exposure to PAHs but not PCB-126 in zebrafish embryos. Comp Biochem Physiol C: Pharmacol Toxicol Endocrinol 155:573–579

    Google Scholar 

  • Geiger GA, Parker SE, Beothy AP, Tucker JA, Mullins MC, Kao GD (2006) Zebrafish as a “biosensor”? Effects of ionizing radiation and amifostine on embryonic viability and development. Cancer Res 66:8172–8181

    Article  Google Scholar 

  • Gilbin R, Alonzo F, Garnier-Laplace J (2008) Effects of chronic external gamma irradiation on growth and reproductive success of Daphnia magna. J Environ Radioact 99:134–145

    Article  Google Scholar 

  • Goldsmith JR, Jobin C (2012) Think small: zebrafish as a model system of human pathology. J Biomed Biotechnol 2012:817341

    Article  Google Scholar 

  • Goth L (1991) A simple method for determination of serum catalase activity and revision of reference range. Clin Chim Acta 196:143–151

    Article  Google Scholar 

  • Halliwell B (2002) Effect of diet on cancer development: is oxidative DNA damage a biomarker? Free Radic Biol Med 32:968–974

    Article  Google Scholar 

  • Hou J, Li L, Xue T, Long M, Su Y, Wu N (2014) Damage and recovery of the ovary in female zebrafish i.p.-injected with MC-LR. Aquat Toxicol 155:110–118

    Article  Google Scholar 

  • Jin Y, Liu Z, Liu F, Ye Y, Peng T, Fu Z (2015) Embryonic exposure to cadmium (II) and chromium (VI) induce behavioral alterations, oxidative stress and immunotoxicity in zebrafish (Danio rerio). Neurotoxicol Teratol 48:9–17

    Article  Google Scholar 

  • Keen JH, Habig WH, Jakoby WB (1976) Mechanism for the several activities of the glutathione S-transferases. J Biol Chem 251:6183–6188

    Google Scholar 

  • Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF (1995) Stages of embryonic development of the zebrafish. Dev Dyn 203:253–310

    Article  Google Scholar 

  • Kumar KB, Kuttan R (2004) Protective effect of an extract of Phyllanthus amarus against radiation-induced damage in mice. J Radiat Res 45:133–139

    Article  Google Scholar 

  • Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410

    Article  Google Scholar 

  • Moreno I, Pichardo S, Jos A, Gómez-Amores L, Mate A, Vazquez CM, Cameán AM (2005) Antioxidant enzyme activity and lipid peroxidation in liver and kidney of rats exposed to microcystin-LR administered intraperitoneally. Toxicon 45:395–402

    Article  Google Scholar 

  • Nair RN, Sunny F, Chopra M, Sharma LK, Puranik VD, Ghosh AK (2014) Estimation of radioactive leakages into the Pacific Ocean due to Fukushima nuclear accident. Environ Earth Sci 71:1007–1019

    Article  Google Scholar 

  • Ng CYP, Kong EY, Konishi T, Kobayashi A, Suya N, Cheng SH, Yu KN (2015a) Low-dose neutron dose response of zebrafish embryos obtained from the Neutron exposure Accelerator System for Biological Effect Experiments (NASBEE) facility. Radiat Phys Chem 114:12–17

    Article  ADS  Google Scholar 

  • Ng CYP, Kong EY, Kobayashi A, Suya N, Uchihori Y, Cheng SH, Konishi T, Yu KN (2015b) Neutron induced bystander effect among zebrafish embryos. Radiat Phys Chem 117:153–159

    Article  ADS  Google Scholar 

  • Nishikimi M (1975) Oxidation of ascorbic acid with superoxide anion generated by the xanthine-xanthine oxidase system. Biochem Biophys Res Commun 63:463–468

    Article  Google Scholar 

  • Pereira S, Bourrachot S, Cavalie I, Plaire D, Dutilleul M, Gilbin R, Adam-Guillermin C (2011) Genotoxicity of acute and chronic gamma-irradiation on zebrafish cells and consequences on embryo development. Environ Toxicol Chem 30:2831–2837

    Article  Google Scholar 

  • Rhee JS, Kim BM, Kang CM, Lee YM, Lee JS (2012) Gamma irradiation-induced oxidative stress and developmental impairment in the hermaphroditic fish, Kryptolebias marmoratus embryo. Environ Toxicol Chem 31:1745–1753

    Article  Google Scholar 

  • Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WJ (1973) Selenium: biochemical role as a component of glutathione peroxidase. Science 179:588–590

    Article  ADS  Google Scholar 

  • Saroya R, Smith R, Seymour C, Mothersill C (2010) Injection of reserpine into zebrafish, prevents fish to fish communication of radiation-induced bystander signals: confirmation in vivo of a role for serotonin in the mechanism. Dose-Response 8:317–330

    Article  Google Scholar 

  • Si J, Zhang H, Wang Z, Wu Z, Lu J, Di C, Zhou X, Wang X (2013) Effects of 12C6+ ion radiation and ferulic acid on the zebrafish (Danio rerio) embryonic oxidative stress response and gene expression. Mutat Res-Fund Mol M 745–746:26–33

    Article  Google Scholar 

  • Simon O, Massarin S, Coppin F, Hinton TG, Gilbin R (2011) Investigating the embryo/larval toxic and genotoxic effects of gamma irradiation on zebrafish eggs. J Environ Radioact 102:1039–1044

    Article  Google Scholar 

  • Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85

    Article  Google Scholar 

  • Srinivasan M, Sudheer AR, Pillai KR, Kumar PR, Sudhakaran PR, Menon VP (2007) Modulatory effects of curcumin on gamma-radiation-induced cellular damage in primary culture of isolated rat hepatocytes. Environ Toxicol Pharmacol 24:98–105

    Article  Google Scholar 

  • Wang C, Smith RW, Duhig J, Prestwich WV, Byun SH, Mcneill FE, Seymour CB, Mothersill CE (2011) Neutrons do not produce a bystander effect in zebrafish irradiated in vivo. Int J Radiat Biol 87:964–973

    Article  Google Scholar 

  • Xia T, Zhao Y, Sager T, George S, Pokhrel S, Li N, Schoenfeld D, Meng H, Lin S, Wang X, Wang M, Ji Z, Zink JI, Mädler L, Castranova V, Lin S, Nel AE (2011) Decreased dissolution of ZnO by iron doping yields nanoparticles with reduced toxicity in the rodent lung and zebrafish embryos. ACS Nano 5:1223–1235

    Article  Google Scholar 

  • Yang L, Ho NY, Alshut R, Legradi J, Weiss C, Reischl M, Mikut R, Liebel U, Müller F, Strähle U (2009) Zebrafish embryos as models for embryotoxic and teratological effects of chemicals. Reprod Toxicol 28:245–253

    Article  Google Scholar 

  • Yuan G, Wang Y, Yuan X, Zhang T, Zhao J, Huang L, Peng S (2014) T-2 toxin induces developmental toxicity and apoptosis in zebrafish embryos. J Environ Sci (China) 26:917–925

    Article  Google Scholar 

  • Yum EHW, Li VWT, Choi VWY, Cheng SH, Yu KN (2010) Effects of alpha particles on zebrafish embryos. Appl Radiat Isot 68:714–717

    Article  Google Scholar 

  • Zhao X, Wang S, Wu Y, You H, Lv L (2013) Acute ZnO nanoparticles exposure induces developmental toxicity, oxidative stress and DNA damage in embryo-larval zebrafish. Aquat Toxicol 136–137:49–59

    Article  Google Scholar 

  • Zhou R, Si J, Zhang H, Wang Z, Li J, Zhou X, Gan L, Liu Y (2014) The effects of X-ray radiation on the eye development of zebrafish. Hum Exp Toxicol 33:1040–1050

    Article  Google Scholar 

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Acknowledgments

This work was supported by the Defense Industrial Technology Development Program (No. B3720132001) and the National Natural Science Foundation of China (No. U1401231).

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    Authors and Affiliations

    1. Key Discipline Laboratory for National Defense for Biotechnology in Uranium Mining and Hydrometallurgy, University of South China, Hengyang, 421001, Hunan Province, China

      Miao Hu, Nan Hu, Dexin Ding, Weichao Zhao, Yongfu Feng, Hui Zhang, Guangyue Li & Yongdong Wang

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    1. Miao Hu
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    2. Nan Hu
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    3. Dexin Ding
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    4. Weichao Zhao
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    Correspondence to Dexin Ding.

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    All applicable international, national and institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of Institutional Animal Care and Use Committees (IACUC) of University of South China, Hengyang, China.

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    Miao Hu and Nan Hu contributed equally to this work.

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    Hu, M., Hu, N., Ding, D. et al. Developmental toxicity and oxidative stress induced by gamma irradiation in zebrafish embryos. Radiat Environ Biophys 55, 441–450 (2016). https://doi.org/10.1007/s00411-016-0663-4

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