CHK1-CENP B/MAD2 is associated with mild oxidative damage-induced sex chromosome aneuploidy of male mouse embryos during in vitro fertilization
CHK1-CENP B/MAD2 is associated with mild oxidative damage-induced sex chromosome aneuploidy of male mouse embryos during in vitro fertilization
复制标题
CHK1-CENP B/MAD2与体外受精过程中轻度氧化损伤诱导的雄性小鼠胚胎性染色体非整倍性相关
DOI：
10.1016/j.freeradbiomed.2019.04.037
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发表时间：
2019-06-01
影响因子：
7.4
通讯作者：
Xiao, Wanfen
中科院分区：
文献类型：
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作者：
Huang, Yue;Hai, Siyao;Xiao, Wanfen
在体外受精（IVF）衍生的胚胎中观察到高发生率的非整倍体，但其形成和修复机制尚不清楚。在这里，我们研究了体外培养条件下产生的活性氧（ROS）的轻微增加对胚胎非整倍体的影响，以及纺锤体组装检查点（SAC）蛋白、有丝分裂抑制缺陷2 （MAD2）和DNA损伤反应（DDR）蛋白、检查点激酶1 （CHK1）在非整倍体修复中的作用。通过DAPI染色、核型分析和下一代测序技术评估染色体异常，我们发现轻度氧化损伤主要通过第一次有丝分裂期间染色体错分离增加雄性小鼠胚胎（41，XXY， + X和41，XYY, + Y）性染色体非整倍体的风险。相对和绝对定量等压标记技术显示，轻度氧化损伤可抑制雄性生殖相关蛋白的表达，包括位于小鼠/人类x染色体上的激酶锚定蛋白4 （AKAP4）。在轻度氧化损伤下，用MAD2抑制剂-1 （M2I-1）或siRNA显微注射CHK1去除MAD2可增加性染色体嵌合率，降低有丝分裂促进因子（MPF）活性。CHK1抑制还降低了着丝粒蛋白B （CENP B）和MAD2的着丝粒定位。这些发现表明，DDR和SAC通过pCHK1 (5345)-CENP B/MAD2-MPF通路参与性染色体嵌合修复；此外，体外受精可能对雄性后代的繁殖能力产生负面影响，这最终取决于它们的持续修复能力。因此，我们认为抗氧化剂，特别是那些靶向改善CHK1-MAD2功能的抗氧化剂，可能是一种有希望的治疗策略，可以减少体外受精胚胎非整倍体的形成，并保持胚胎和后代的基因组完整性。
A high incidence of aneuploidy is observed in vitro fertilization (IVF)-derived embryos, but the formation and repair mechanisms are unknown. Here, we investigated the effects of slightly increased reactive oxygen species (ROS) produced by in vitro culture conditions on embryo aneuploidy and the roles of the spindle assembly checkpoint (SAC) protein, mitotic arrest-deficient 2 (MAD2), and the DNA damage response (DDR) protein, checkpoint kinase 1 (CHK1), in aneuploidy repair. By assessing chromosome abnormalities via DAPI staining, karyotype analysis and next-generation sequencing technology, we demonstrated that mild oxidative damage mainly increased the risk of sex chromosome aneuploidy in male mouse embryos (41,XXY, + X and 41,XYY, + Y) through chromosome mis-segregation during the first mitosis. Isobaric tags for relative and absolute quantitation technology revealed that mild oxidative damage inhibited the expression of male reproduction-related proteins, including a kinase anchor protein 4 (AKAP4), whose gene is located on mouse/human Chromosome X. Under mild oxidative damage, abrogation of MAD2 by MAD2 inhibitor-1 (M2I-1) or CHK1 by siRNA microinjection increased sex chromosome mosaicism rate and reduced mitosis-promoting factor (MPF) activity. CHK1 inhibition also reduced kinetochore localization of centromere protein B (CENP B) and MAD2. These findings show that DDR and SAC are responsible for repair of sex chromosome mosaicism via the pCHK1 (5345)-CENP B/MAD2-MPF pathway; further, IVF may have negative effects on male offspring's reproduction ability, which ultimately depends on their continued repair capability. Therefore, we suggest that antioxidants, especially those targeting improved CHK1-MAD2 function, may be a promising therapeutic strategy to reduce aneuploidy formation of IVF-derived embryos and to maintain genome integrity of embryo and offspring.