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Evaluation of a modified comet assay to detect DNA damage in mammalian sperm exposed in vitro to different mutagenic compounds

TitoloEvaluation of a modified comet assay to detect DNA damage in mammalian sperm exposed in vitro to different mutagenic compounds
Tipo di pubblicazioneArticolo su Rivista peer-reviewed
Anno di Pubblicazione2010
AutoriVillani, Paola, Spanò M., Pacchierotti Francesca, Weimer M., and Cordelli Eugenia
RivistaReproductive Toxicology
Volume30
Paginazione44-49
ISSN08906238
Parole chiaveanimal, animal cell, Animals, article, Bleomycin, Bovinae, bull (bovine), Cattle, colchicine, Comet Assay, concentration response, controlled study, diethyl sulfate, DNA, DNA damage, dose response, Dose-Response Relationship, Drug, drug effects, genotoxicity, HeLa cell, HeLa cell line, Hela Cells, human, Humans, in vitro study, male, Mammalia, metabolism, mitomycin C, mutagenic agent, Mutagens, nonhuman, procedures, Reliability, reproducibility, Reproducibility of Results, sperm, Spermatozoa, spermatozoon, standards, Toxicity
Abstract

The final stages of male gametogenesis are sensitive targets of DNA-reactive chemicals, most of which form adducts. Comet assay is a widely applied genotoxicity test that reveals DNA adducts through breaks formed during repair processes. However, sperm cells are essentially devoid of repair enzymes and comet assay is poorly sensitive in detecting chemically induced DNA lesions in sperm. To overcome such limitation, in a previous paper we proposed a modified protocol for comet assay. In this work we further tested the method treating bull sperm with additional mutagens (diethylsulfate, mitomycin C, bleomycin and colchicine) in parallel with the standard comet assay. No treatment-related increase of DNA migration was ever detected with the standard protocol. A dose-dependent effect of diethylsulfate, was obtained with the modified assay. As expected, the mitotic poison colchicine resulted negative even by the modified assay. Results with the other two compounds were consistent with their mechanism of action. © 2009 Elsevier Inc.

Note

cited By 9

URLhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-77954087988&doi=10.1016%2fj.reprotox.2009.10.015&partnerID=40&md5=537d1765f5b38e387e0b226272153045
DOI10.1016/j.reprotox.2009.10.015
Citation KeyVillani201044