Naslov
Zebrafish genome stability after exposure to model genotoxicants

Autori
Šrut Maja ; Štambuk Anamaria ; Bourdineaud Jean-Paul ; Klobučar Göran

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Comparative Biochemistry and Physiology / Mommsen, T.P. ; Walsh, P.J. - Bilbao, 2012, 15-15

Skup
28th Congress-European Society for Comparative Physiology and Biochemistry

Mjesto i datum
Bilbao, Španjolska, 02.09.2012. - 05.09.2012

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Danio rerio; genotoxicity; Comet assay; AFLP; RAPD

Sažetak
Sublethal exposure to a variety of genotoxic agents in the environment may directly or indirectly influence genome integrity of exposed organisms and for that reason various methods for detection of DNA damage were developed. In this study we have tested and compared three different methods for detection of DNA damage and alterations using in vivo and in vitro zebrafish models. The exposure of embryonic cell line (PAC-2) and zebrafish embryos to genotoxic agents (benzo[a]pyrene (B[a]P) and ethyl methanesulfonate (EMS)) was carried out for 6 days followed by 6 days of recovery period. Genome stability of tested models was assessed using Comet assay, quantitative RAPD (random amplified polymorphic DNA) and AFLP (amplified fragment length polymorphism). Comet assay revealed significant increase of DNA damage in zebrafish embryos following both toxicant treatments and this damage remained unrepaired after the recovery period. However, in PAC-2 cell line significant increase of DNA damage was observed only upon EMS treatment. RAPD showed significant increase in number of hybridization sites in DNA of zebrafish embryos and decrease in DNA of PAC-2 cell line upon exposure to lower B[a]P concentration, while AFLP revealed changes in profiles, such as loss or appearance of fragments as well as changes in fragment amplification intensity, following both toxicants treatment. Our results indicate that RAPD and AFLP are sensitive methods for detection of DNA alterations since they showed changes in the genome even when Comet assay indicated lack of significant damage. This could be due to the presence of more persistent DNA changes such as formation of DNA adducts, mutations or conformational and structural changes in the DNA, all undetected in the Comet assay. These results thus encourage the simultaneous use of various methods and models for revealing the consequences of genotoxic insult and suggest that quantitative RAPD and AFLP could be useful biomarkers in such surveys.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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