| Autor: |
Nguyen, Hieu, Ribas-Maynou, Jordi, Wu, Hongwen, Quon, Beverly, Inouye, Tracy, Walker, Brienne, Langaman, Caitlin, Huang, Thomas T F, Ward, W Steven |
| Zdroj: |
Biology of Reproduction; November 2023, Vol. 109 Issue: 5 p635-643, 9p |
| Abstrakt: |
We previously demonstrated that MnCl2induces double-stranded DNA breaks in sperm in a process that we term as sperm chromatin fragmentation. Here, we tested if the levels of double-stranded DNA breaks were corelated to the concentration of MnCl2, and we compared this to another agent that causes single-stranded DNA breaks, H2O2. We found that both methods have the advantage of inducing DNA breaks in a concentration-dependent manner. Mouse sperm were treated with varying concentrations of either H2O2or MnCl2, and the DNA damage was assessed by pulse-field gel electrophoresis, and the alkaline and neutral comet assays. Oocytes were injected with either treated sperm and the resulting embryos analyzed with an embryoscope to detect subtle changes in embryonic development. We confirmed that H2O2treatment induced primarily single-stranded DNA breaks and MnCl2induced primarily double-stranded DNA breaks, indicating different mechanisms of damage. These sperm were injected into oocytes, and the development of the resulting embryos followed with an embryoscope equipped with time lapse recording. We found that aberrations in early embryonic development by day 2 with even the lowest levels of DNA damage and that the levels of embryonic aberrations correlated to the concentration of either H2O2or MnCl2. Low levels of H2O2caused significantly more aberrations in embryonic development than low levels of MnCl2even though the levels of DNA damage as measured by comet assays were similar. These data demonstrate that even low levels of sperm DNA damage cause delays and arrests in embryonic development.Treatment of mouse sperm with doses of DNA damaging agents that are too low to cause easily visualized DNA breaks cause significant embryonic development problems when used to fertilize untreated eggs.Graphical Abstract |
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