Interpreting sperm DNA damage in a diverse range of mammalian sperm by means of the two-tailed comet assay

Cortés-Gutiérrez, Elva I., López-Fernández, Carmen, Fernández, José Luis, Dávila-Rodríguez, Martha I., Johnston, Stephen D. and Gosálvez, Jaime (2014) Interpreting sperm DNA damage in a diverse range of mammalian sperm by means of the two-tailed comet assay. Frontiers in Genetics, 5 NOV: 404.1-404.11. doi:10.3389/fgene.2014.00404

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Author Cortés-Gutiérrez, Elva I.
López-Fernández, Carmen
Fernández, José Luis
Dávila-Rodríguez, Martha I.
Johnston, Stephen D.
Gosálvez, Jaime
Title Interpreting sperm DNA damage in a diverse range of mammalian sperm by means of the two-tailed comet assay
Journal name Frontiers in Genetics   Check publisher's open access policy
ISSN 1664-8021
Publication date 2014-11-27
Sub-type Critical review of research, literature review, critical commentary
DOI 10.3389/fgene.2014.00404
Open Access Status DOI
Volume 5
Issue NOV
Start page 404.1
End page 404.11
Total pages 11
Place of publication Lausanne, Switzerland
Publisher Frontiers Research Foundation
Language eng
Subject 1313 Molecular Medicine
1311 Genetics
2716 Genetics (clinical)
Abstract Sperm DNA damage is associated with fertilization failure, impaired pre-and post- embryo implantation and poor pregnancy outcome. A series of methodologies to assess DNA damage in spermatozoa have been developed but most are unable to differentiate between single-stranded DNA breaks (SSBs) and double-stranded DNA breaks (DSBs) on the same sperm cell. The two-dimensional Two-Tailed Comet assay (TT-comet) protocol highlighted in this review overcomes this limitation and emphasizes the importance in accounting for the difference in sperm protamine composition at a species-specific level for the appropriate preparation of the assay. The TT-comet is a modification of the original comet assay that uses a two dimensional electrophoresis to allow for the simultaneous evaluation of DSBs and SSBs in mammalian spermatozoa. Here we have compiled a retrospective overview of how the TT-comet assay has been used to investigate the structure and function of sperm DNA across a diverse range of mammalian species (eutheria, metatheria, and prototheria). When conducted as part of the TT-comet assay, we illustrate (a) how the alkaline comet single assay has been used to help understand the constitutive and transient changes in DNA structure associated with chromatin packing, (b) the capacity of the TT-comet to differentiate between the presence of SSBs and DSBs (c) and the possible implications of SSBs or DSBs for the assessment of infertility.
Keyword Sperm DNA damage
Fertility
Male factor
Mammalian reproduction
Eutheria
Metatheria
Prototheria
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Critical review of research, literature review, critical commentary
Collections: School of Agriculture and Food Sciences
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Citation counts: TR Web of Science Citation Count  Cited 9 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 12 times in Scopus Article | Citations
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Created: Sat, 06 Dec 2014, 21:27:37 EST by Associate Professor Stephen Johnston on behalf of School of Agriculture and Food Sciences