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

Overview of attention for article published in Frontiers in Genetics, November 2014
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Title
Interpreting sperm DNA damage in a diverse range of mammalian sperm by means of the two-tailed comet assay
Published in
Frontiers in Genetics, November 2014
DOI 10.3389/fgene.2014.00404
Pubmed ID
Authors

Elva I. Cortés-Gutiérrez, Carmen López-Fernández, José Luis Fernández, Martha I. Dávila-Rodríguez, Stephen D. Johnston, Jaime Gosálvez

Abstract

Key ConceptsThe two-dimensional Two-Tailed Comet assay (TT-comet) protocol is a valuable technique to differentiate between single-stranded (SSBs) and double-stranded DNA breaks (DSBs) on the same sperm cell.Protein lysis inherent with the TT-comet protocol accounts for differences in sperm protamine composition at a species-specific level to produce reliable visualization of sperm DNA damage.Alkaline treatment may break the sugar-phosphate backbone in abasic sites or at sites with deoxyribose damage, transforming these lesions into DNA breaks that are also converted into ssDNA. These lesions are known as Alkali Labile Sites "ALSs."DBD-FISH permits the in situ visualization of DNA breaks, abasic sites or alkaline-sensitive DNA regions.The alkaline comet single assay reveals that all mammalian species display constitutive ALS related with the requirement of the sperm to undergo transient changes in DNA structure linked with chromatin packing.Sperm DNA damage is associated with fertilization failure, impaired pre-and post- embryo implantation and poor pregnancy outcome.The TT is a valuable tool for identifying SSBs or DSBs in sperm cells with DNA fragmentation and can be therefore used for the purposes of fertility assessment. 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.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 76 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Brazil 1 1%
Unknown 75 99%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 16 21%
Researcher 12 16%
Student > Bachelor 9 12%
Student > Master 8 11%
Student > Doctoral Student 4 5%
Other 9 12%
Unknown 18 24%
Readers by discipline Count As %
Agricultural and Biological Sciences 18 24%
Medicine and Dentistry 13 17%
Biochemistry, Genetics and Molecular Biology 7 9%
Environmental Science 5 7%
Pharmacology, Toxicology and Pharmaceutical Science 4 5%
Other 10 13%
Unknown 19 25%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 03 September 2015.
All research outputs
#14,204,846
of 22,771,140 outputs
Outputs from Frontiers in Genetics
#3,914
of 11,758 outputs
Outputs of similar age
#192,013
of 361,861 outputs
Outputs of similar age from Frontiers in Genetics
#68
of 109 outputs
Altmetric has tracked 22,771,140 research outputs across all sources so far. This one is in the 35th percentile – i.e., 35% of other outputs scored the same or lower than it.
So far Altmetric has tracked 11,758 research outputs from this source. They receive a mean Attention Score of 3.7. This one has gotten more attention than average, scoring higher than 62% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 361,861 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 44th percentile – i.e., 44% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 109 others from the same source and published within six weeks on either side of this one. This one is in the 33rd percentile – i.e., 33% of its contemporaries scored the same or lower than it.