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Zebrafish genome instability after exposure to model genotoxicants

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Abstract

Sublethal exposure to environmental genotoxicants may impact genome integrity in affected organisms. It is therefore necessary to develop tools to measure the extent and longevity of genotoxicant-induced DNA damage, and choose appropriate model organisms for biomonitoring. To this end, markers of DNA damage were measured in zebrafish larvae and adults following exposure to model genotoxicants (benzo[a]pyrene and ethyl methanesulfonate). Specifically, we assessed primary DNA damage and the existence of potentially persistent genomic alterations through application of the comet assay, quantitative random amplified polymorphic DNA (qRAPD) and amplified fragment length polymorphism (AFLP) assays. Furthermore, expression of genes involved in DNA repair, oxidative stress response and xenobiotic metabolism was evaluated as well. Additionally, the AFLP method was applied to adult specimens 1 year after larval exposure to the genotoxicants to evaluate the longevity of the observed DNA alterations. Large numbers of DNA alterations were detected in larval DNA using the comet assay, qRAPD and AFLP, demonstrating that zebrafish larvae are a sensitive model for revealing genotoxic effects. Furthermore, some of these genomic alterations persisted into adulthood, indicating the formation of stable genomic modifications. qRAPD and AFLP methods proved to be highly sensitive to genotoxic effects, even in cases when the comet assay indicated a lack of significant damage. These results thus support the use of zebrafish larvae as a sensitive model for monitoring the impact of genotoxic insult and give evidence of the longevity of genomic modifications induced by genotoxic agents.

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Acknowledgments

The authors are grateful to the students Antonio Karaga and Aleksandar Lazić for help with the preparation and analysis of comet assay slides and to Adam Maguire for English revision of the manuscript. This study was part of a Ph.D. thesis and was conducted within the framework of Project No. 119-0982934-3110 supported by the Ministry of Science, Education and Sports of the Republic of Croatia and of program Tractifs No. ANR-07-SEST-02301 supported by the French National Research Agency. Maja Šrut was awarded a scholarship by the Croatian Science Foundation to perform a portion of the research at the University of Bordeaux, France.

Ethical standards

Experiments performed in this study comply with the current laws of Croatia.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Zoology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10000, Zagreb, Croatia

    Maja Šrut, Anamaria Štambuk & Göran I. V. Klobučar

  2. Arcachon Marine Station, CNRS, UMR EPOC 5805, University of Bordeaux, Place du Docteur Peyneau, 33120, Arcachon, France

    Jean-Paul Bourdineaud

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  1. Maja Šrut
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  2. Anamaria Štambuk
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  3. Jean-Paul Bourdineaud
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  4. Göran I. V. Klobučar
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Correspondence to Maja Šrut.

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Šrut, M., Štambuk, A., Bourdineaud, JP. et al. Zebrafish genome instability after exposure to model genotoxicants. Ecotoxicology 24, 887–902 (2015). https://doi.org/10.1007/s10646-015-1432-x

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