The Comet assay for detection of <scp>DNA</scp> damage in canine sperm

Pereira, AF; Borges, Paulo; Fontbonne, Alain; Cardoso, Luı́s; Gaivão, Isabel; Martins‐Bessa, Ana

doi:10.1111/rda.13042

Cited by 12 publications

(6 citation statements)

References 9 publications

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“…Dogs from different regions of the city of São Paulo, Brazil were used to evaluate the extent of DNA damage in the olfactory and respiratory epithelia, indicating increased DNA-damaging effects in relation to environmental factors [397]. Besides, the comet assay is also used for detection of DNA damage in canine sperm [398].…”

Section: Domestic Mammalsmentioning

confidence: 99%

The comet assay in animal models: From bugs to whales – (Part 2 Vertebrates)

Gajski

Žegura

Ladeira

et al. 2019

Mutation Research/Reviews in Mutation Research

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The comet assay has become one of the methods of choice for the evaluation and measurement of DNA damage. It is sensitive, quick to perform and relatively affordable for the evaluation of DNA damage and repair at the level of individual cells. The comet assay can be applied to virtually any cell type derived from different organs and tissues. Even though the comet assay is predominantly used on human cells, the application of the assay for the evaluation of DNA damage in yeast, plant and animal cells is also quite high, especially in terms of biomonitoring. The present extensive overview on the usage of the comet assay in animal models will cover both terrestrial and water environments. The first part of the review was focused on studies describing the comet assay applied in invertebrates. The second part of the review, (Part 2) will discuss the application of the comet assay in vertebrates covering cyclostomata, fishes, amphibians, reptiles, birds and mammals, in addition to chordates that are regarded as a transitional form towards vertebrates. Besides numerous vertebrate species, the assay is also performed on a range of cells, which includes blood, liver, kidney, brain, gill, bone marrow and sperm cells. These cells are readily used for the evaluation of a wide spectrum of genotoxic agents both in vitro and in vivo. Moreover, the use of vertebrate models and their role in environmental biomonitoring will also be discussed as well as the comparison of the use of the comet assay in vertebrate and human models in line with ethical principles. Although the comet assay in vertebrates is most commonly used in laboratory animals such as mice, rats and lately zebrafish, this paper will only briefly review its use regarding laboratory animal models and rather give special emphasis to the increasing usage of the assay in domestic and wildlife animals as well as in various ecotoxicological studies.

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Section: Domestic Mammalsmentioning

confidence: 99%

The comet assay in animal models: From bugs to whales – (Part 2 Vertebrates)

Gajski

Žegura

Ladeira

et al. 2019

Mutation Research/Reviews in Mutation Research

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“…As the sperm head consists almost entirely of DNA, it has been hypothesized that sperm head morphometric parameters may reflect chromatin organization (Lange‐Consiglio et al., 2010). Studies have been carried out in dogs that relate DNA damage to sperm head morphometry (Núñez‐Martinez et al, 2005; Lange‐Consiglio et al., 2010; Urbano et al., 2017); but few studies evaluate the association between DNA damage and other routine sperm parameters (Pereira et al., 2017; Prinosilova et al., 2012).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of DNA fragmentation in dog sperm using the sperm chromatin dispersion test

Monachesi

Gallelli

Neild

et al. 2022

Reprod Domestic Animals

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The study's objective was to adapt the Sperm Chromatin Dispersion (SCD) protocol to evaluate sperm DNA fragmentation and implement a fragmentation control in dogs. Correlation between DNA status and routine sperm parameters was also analysed. To adapt the SCD, two different mercaptoethanol (ME) concentrations were assayed (2.5% and 5%) in fourteen ejaculates from seven dogs and semen incubation with 0.3 M NaOH for 15 min at room temperature was assayed as a control for sperm DNA fragmentation. Data were analysed using a Mann–Whitney test and either Pearson's or Spearman's correlation. The selected ME concentration to use in the SCD test was 5%, as it produced the largest DNA dispersion halo while preserving the core nucleus structure. Four DNA halo patterns were identified as follows: large dispersion halos, medium halos, small halos and nuclei without halos. Semen incubated with NaOH showed 100% sperm without halos (damaged DNA). A significant positive correlation was observed between sperm with fragmented DNA and sperm with coiled tails. Thus, it was possible to adapt the SCD protocol to evaluate dog sperm DNA fragmentation in raw semen without using a commercial kit and establish incubation with NaOH as a DNA fragmentation control. Only coiled tails showed correlation with DNA fragmentation.

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“…The most frequently applied methodology for determining induction of DNA damage and repair processes in isolated cells is the alkaline SCGE, which can be used in a multiplicity of experimental models (Nikoloff et al 2014;Pereira et al 2017;Laborde et al 2020). For instance, Padula et al (2012) found increased DNA damage in CHO-K1 cells treated with 3.75 μg/mL AMZ for 16 h; Radakovic et al (2013) demonstrated that various AMZ concentrations (0.035, 0.35, 3.5, 35, and 350 μg/mL) induced DNA damage in human lymphocytes, finding the highest DNA damage with 3.5 μg/ mL while the highest concentrations of AMZ (35 and 350 μg/ mL) showed lower levels of DNA damage, they suggest that this is possibly due to formation of crosslinks.…”

Section: Discussionmentioning

confidence: 99%

Amitraz induced cytotoxic effect on bovine cumulus cells and impaired oocyte maturation

Nikoloff

Martín

Fabra

et al. 2021

Environ Sci Pollut Res

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The aim of this study was to evaluate the genotoxic and cytotoxic effects of amitraz (AMZ) on the primary culture of bovine cumulus cells (CC) and oocyte nuclear maturation. Cytotoxicity was evaluated by assessing mitochondrial activity with the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Genotoxicity was estimated using the alkaline single cell gel electrophoresis (SCGE) assay. Apoptosis was detected with the Annexin V-affinity assay. The in vitro maturation test was performed in bovine oocytes. To understand AMZ action, glutathione content, superoxide dismutase enzyme activity, and lipid peroxidation were evaluated in CC. Results showed that AMZ lethal concentration (LC 50 24h ) for bovine CC was 32.55 μg/mL (MTT assay). A 25 μg/mL induced late apoptosis and necrotic cells (p < 0.05); however, DNA damage was decreased at the same concentration (SCGE assay; p < 0.05). A decrease in metaphase II was observed at 25 μg/mL, and degenerate oocytes were observed at 15 and 25 μg/mL (p < 0.05). None of the oxidative stress parameters evaluated showed significant differences. This study contributes to a better understanding of AMZ in this model, suggesting its potential cytotoxicity and impact on bovine reproduction.

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The Comet assay for detection of DNA damage in canine sperm

Cited by 12 publications

References 9 publications

The comet assay in animal models: From bugs to whales – (Part 2 Vertebrates)

The comet assay in animal models: From bugs to whales – (Part 2 Vertebrates)

Evaluation of DNA fragmentation in dog sperm using the sperm chromatin dispersion test

Amitraz induced cytotoxic effect on bovine cumulus cells and impaired oocyte maturation

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