The Comet Assay in Toxicology
Chapter 15: The Comet Assay in Sperm—Assessing Genotoxins in Male Germ Cells
Published:07 Oct 2016
Special Collection: 2016 ebook collectionSeries: Issues in Toxicology
A. Baumgartner, A. Ali, C. Normington, and D. Anderson, in The Comet Assay in Toxicology, ed. D. Anderson and A. Dhawan, The Royal Society of Chemistry, 2nd edn, 2016, ch. 15, pp. 390-456.
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In a modern society, the environment as well as the lifestyles we choose may expose us to genotoxic compounds that may not only damage somatic cells but also our germ cells. As these reproductive cells pass genetic material on to the next generations their DNA integrity is of crucial importance. In recent years, it become clear that a strong link between paternal smoking and the induction of DNA damage in the unexposed offspring exists. Such compromised DNA can then be transmitted via the spermatozoal genome to the egg causing gene mutations in the offspring. The Comet or single-cell gel electrophoresis (SCGE) assay is a rapid, sensitive and reliable method to detect DNA damage and to assess the integrity of the genome within single mature male germ cells. This chapter will provide an overview of the use of the in vivo and in vitro Comet assay utilising sperm or testicular cells in reproductive toxicology. This includes considerations of damage assessed by protocol modification, cryopreservation versus the use of fresh sperm, viability and statistics. The use of sperm to assess possible detrimental effects on germ cell DNA might play a crucial role in investigating reprotoxins, which are harmful to humans, but may also contribute to areas such as aquatic and terrestrial ecotoxicology. The sperm Comet assay allows reliable in vitro and in vivo assessments of various environmental and lifestyle genotoxins, presumed or proven to damage the DNA.