ArticleMolecular human reproduction2014
Double-stranded DNA breaks hidden in the neutral Comet assay suggest a role of the sperm nuclear matrix in DNA integrity maintenance.
Article in Molecular human reproduction, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers, 1 of them a synthesis that pooled it.
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Who cites it
21 citing papers in PubMed, 1 synthesis or guideline pooled it.
- The use of follicle stimulating hormone (FSH) for the treatment of the infertile man: position statement from the Italian Society of Andrology and Sexual Medicine (SIAMS).Journal of endocrinological investigation · 2018Guideline
- Influence of pH and lysis duration on DNA damage detection: a comparison of neutral and alkaline comet assays.Mutagenesis · 2025Article
- Low levels of mouse sperm chromatin fragmentation delay embryo development.Biology of reproduction · 2023Article
- Can a sperm selection technique improve embryo ploidy?Andrology · 2023Article
- An intracellular, non-oxidative factor activates in vitro chromatin fragmentation in pig sperm.Biological research · 2023Article
- Sperm DNA integrity does play a crucial role for embryo development after ICSI, notably when good-quality oocytes from young donors are used.Biological research · 2022Article
- Sperm degradation after vasectomy follows a sperm chromatin fragmentation-dependent mechanism causing DNA breaks in the toroid linker regions.Molecular human reproduction · 2022Article
- Functional Aspects of Sperm Chromatin Organization.Results and problems in cell differentiation · 2022Review
- PARP-1 overexpression does not protect HaCaT cells from DNA damage induced by SiOToxicology research · 2021Article
- Association of Zinc deficiency, oxidative stress and increased double-stranded DNA breaks in globozoospermic infertile patients and its implication for the assisted reproductive technique.Translational andrology and urology · 2021Article
- Magnetic-Activated Cell Sorting (MACS): A Useful Sperm-Selection Technique in Cases of High Levels of Sperm DNA Fragmentation.Journal of clinical medicine · 2020Article
- Single and Double Strand Sperm DNA Damage: Different Reproductive Effects on Male Fertility.Genes · 2019Review
- General control non-repressible 20 (GCN20) functions in root growth by modulating DNA damage repair in Arabidopsis.BMC plant biology · 2018Article
- Probing the Origins of 1,800 MHz Radio Frequency Electromagnetic Radiation Induced Damage in Mouse Immortalized Germ Cells and SpermatozoaFrontiers in public health · 2018Article
- Fatherhood and Sperm DNA Damage in Testicular Cancer Patients.Frontiers in endocrinology · 2018Review
- Eight tests for sperm DNA fragmentation and their roles in the clinic.Translational andrology and urology · 2017Article
- Luminal fluid of epididymis and vas deferens contributes to sperm chromatin fragmentation.Human reproduction (Oxford, England) · 2015Article
- A simple sperm DNA toroid integrity test and risk of miscarriage.BioMed research international · 2015Article
- Medical school hotline: The Institute for Biogenesis Research: a flower in the Pacific.Hawai'i journal of medicine & public health : a journal of Asia Pacific Medicine & Public Health · 2014Article
- A model for the control of DNA integrity by the sperm nuclear matrix.Asian journal of andrologyReview
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4 authors.
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Abstract
We used a mouse model in which sperm DNA damage was induced to understand the relationship of double-stranded DNA (dsDNA) breaks to sperm chromatin structure and to the Comet assay. Sperm chromatin fragmentation (SCF) produces dsDNA breaks located on the matrix attachment regions, between protamine toroids. In this model, epididymal sperm induced to undergo SCF can religate dsDNA breaks while vas deferens sperm cannot. Here, we demonstrated that the conventional neutral Comet assay underestimates the epididymal SCF breaks because the broken DNA ends remain attached to the nuclear matrix, causing the DNA to remain associated with the dispersion halo, and the Comet tails to be weak. Therefore, we term these hidden dsDNA breaks. When the Comet assay was modified to include an additional incubation with sodium dodecyl sulfate (SDS) and dithiothreitol (DTT) after the conventional lysis, thereby solubilizing the nuclear matrix, the broken DNA was released from the matrix, which resulted in a reduction of the sperm head halo and an increase in the Comet tail length, exposing the hidden dsDNA breaks. Conversely, SCF-induced vas deferens sperm had small halos and long tails with the conventional neutral Comet assay, suggesting that the broken DNA ends were not tethered to the nuclear matrix. These results suggest that the attachment to the nuclear matrix is crucial for the religation of SCF-induced DNA breaks in sperm. Our data suggest that the neutral Comet assay identifies only dsDNA breaks that are released from the nuclear matrix and that the addition of an SDS treatment can reveal these hidden dsDNA breaks.
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