Oxidative Stress Delays Prometaphase/Metaphase of the First Cleavage in Mouse Zygotes via the MAD2L1-Mediated Spindle Assembly Checkpoint.

Oxidative Stress Delays Prometaphase/Metaphase of the First Cleavage in Mouse Zygotes via the MAD2L1-Mediated Spindle Assembly Checkpoint.
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氧化应激通过 MAD2L1 介导的纺锤体组装检查点延迟小鼠受精卵第一次分裂的前中期/中期

DOI:
10.1155/2017/2103190
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发表时间:
2017
影响因子:
--
通讯作者:
Wang B
Wang B
中科院分区:
生物学2区
文献类型:
--
作者:
Wu Q;Li Z;Huang Y;Qian D;Chen M;Xiao W;Wang B

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在受精卵中,DNA损伤会推迟第一次卵裂,从而使修复成为可能。我们先前的研究发现,0.03 mM的过氧化氢(H_2O_2)是诱导小鼠受精卵DNA氧化损伤的最低浓度,这与临床现象最相似。在这项研究中,我们量化了不同发育阶段囊胚细胞的卵裂率,然后用免疫荧光检测受精卵中γ-H_2A组蛋白家族成员X(DNA损伤的标志)的激活,以证实H_2O_2处理后的受精卵发生了氧化损伤。在受精后不同时间监测H3S10P(组蛋白H3上Ser10的磷酸化)水平,发现0.03 mM的H_2O_2处理受精卵导致了早中期/中期的延迟。此外,对纺锤体组装检查点(SAC)中的有丝分裂停滞缺陷2-样1和蛋白激酶TTK的免疫荧光染色表明,这种延迟可能与SAC的激活有关。这些研究氧化应激与卵泡刺激素的关系有助于提高体外受精的成功率。
In zygotes, DNA damage delays the first cleavage to enable repair. Our previous study found that 0.03 mM hydrogen peroxide (H2O2) was the minimum concentration required for induction of oxidative DNA damage in mouse zygotes and that this represented the most similar situation to the clinical phenomenon. In this study, we quantified the cleavage rates of cells in blastocysts at different developmental stages, followed by immunofluorescence to detect activation of γ‐H2A histone family member X (a marker of DNA damage) in zygotes to confirm that oxidative DNA damage was induced in H2O2‐treated zygotes. Monitoring H3S10P (phosphorylation of Ser10 on histone H3; a prometaphase/metaphase marker) levels at different hour postinsemination revealed that treatment of zygotes with 0.03 mM H2O2 resulted in a prometaphase/metaphase delay. Furthermore, immunofluorescence staining for mitotic arrest deficient 2‐like 1 and the protein kinase TTK, components of the spindle assembly checkpoint (SAC), suggested that this delay possibly involved SAC activation. These studies of the relationships between oxidative stress and SAC can promote the success rate of in vitro fertilization.
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