Zygotic G2/M cell cycle arrest induced by ATM/Chk1 activation and DNA repair in mouse embryos fertilized with hydrogen peroxide-treated epididymal mouse sperm.
Zygotic G2/M cell cycle arrest induced by ATM/Chk1 activation and DNA repair in mouse embryos fertilized with hydrogen peroxide-treated epididymal mouse sperm.
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用过氧化氢处理的附睾小鼠精子受精的小鼠胚胎中 ATM/Chk1 激活和 DNA 修复诱导的合子 G2/M 细胞周期停滞
DOI：
10.1371/journal.pone.0073987
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Wang X
中科院分区：
文献类型：
--
作者：
Wang B;Li Z;Wang C;Chen M;Xiao J;Wu X;Xiao W;Song Y;Wang X
用于辅助生殖的人类精子冷冻保存受到ROS诱导的精子冷冻损伤的影响。我们以前的模型研究中，小鼠精子用过氧化氢处理，以模拟精子DNA损伤引起的冷冻保存诱导的活性氧发现，小鼠胚胎受精与处理的精子表现出延迟卵裂，可能与细胞周期阻滞。延迟背后的DNA损伤检查点通路仍然难以捉摸。此外，我们以前的研究也表明，DNA损伤修复标记物γ H2 AX在类似受精的小鼠胚胎中是有功能的，但其完整性和正确性尚不清楚，需要更多的研究，因为DNA修复的完整性和正确性不足会引发细胞凋亡。基于上述模型，我们采用胚胎培养、倒置显微镜、BrdU掺入和免疫荧光等方法，探讨了H2 O2处理精子受精的小鼠受精卵细胞发生细胞周期阻滞的时相和潜在的DNA损伤检查点通路。我们还采用Tunel技术研究了不同发育阶段相似受精的小鼠胚胎的凋亡情况，以验证精子DNA损伤修复的完整性和正确性。我们发现G2/M期细胞周期阻滞受精卵与H2 O2处理的精子。在处理组的合子中检测到ATM（pSer-1981）和Chk 1（pSer-345）激活，而不是ATR（pSer-428）和Chk 2（pThr-68）激活。处理组不同发育阶段胚胎的凋亡率与未处理组无明显差异。结论：ATM（pSer-1981）-Chk 1（pSer-345）级联反应可能介导了氧应激精子受精的小鼠受精卵细胞G2/M期阻滞，并为精子DNA损伤修复提供了时间，且DNA损伤修复可能是有效的。
Human sperm cryopreservation for assisted reproduction is compromised by ROS-induced sperm cryodamage. Our previous model study in which mouse sperm were treated with H2O2 to simulate sperm DNA-damage caused by cryopreservation-induced ROS have discovered that mouse embryos fertilized with treated sperm showed a delay in cleavage that might be associated with cell cycle arrest. The DNA-damage checkpoint pathway underlying the delay remained elusive. Moreover, our previous study have also indicated that γH2AX, the DNA-damage repair marker, was functional in mouse embryos similarly fertilized, but the completeness and correctness are unknown and warrant more studies because insufficiency of completeness and correctness of DNA repair would otherwise trigger apoptosis. Based on the aforementioned model, we used embryo culture, inverted microscope, BrdU incorporation and immunofluorescence to explore the cell cycle phase that arrest occurred and the underlying DNA-damage checkpoint pathway in mouse zygotes fertilized with H2O2-treated sperm. We also adopted Tunel to investigate the apoptosis of mouse embryos similarly fertilized at different developmental stages to testify the completeness and correctness of sperm-derived DNA-damage repair. We found G2/M cell cycle arrest in zygotes fertilized with H2O2-treated sperm. ATM (pSer-1981) and Chk1 (pSer-345) activations, rather than ATR (pSer-428) and Chk2 (pThr-68), were detected in zygotes of the treated group. The apoptosis of embryos of different developmental stages of the treated group weren’t different from those of the untreated group. In conclusions, ATM (pSer-1981)-Chk1 (pSer-345) cascade might have mediated G2/M cell cycle arrest and allowed time to facilitate sperm-derived DNA-damage repair in mouse zygotes fertilized with oxygen-stressed sperm, and the DNA-damage repair might be effective.
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影响因子：
6.1
作者：
O'Connell, M;McClure, N;Lewis, SEM
通讯作者：
Lewis, SEM
抗坏血酸和过氧化氢酶对人精子冷冻保存的保护作用
影响因子：
--
作者：
Li, Zhiling;Lin, Qionglin;Liu, Wanmin
通讯作者：
Liu, Wanmin
影响因子：
2.5
作者：
Chatterjee, S;Gagnon, C
通讯作者：
Gagnon, C
影响因子：
56.9
作者：
BRANDRIFF, B;PEDERSEN, RA
通讯作者：
PEDERSEN, RA
影响因子：
2.5
作者：
Liu, SZ;Jiang, MX;Chen, DY
通讯作者：
Chen, DY