Sirt3 protects in vitro-fertilized mouse preimplantation embryos against oxidative stress-induced p53-mediated developmental arrest
Sirt3 protects in vitro-fertilized mouse preimplantation embryos against oxidative stress-induced p53-mediated developmental arrest
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DOI：
10.1172/jci42020
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发表时间：
2010-08-01
影响因子：
15.9
通讯作者：
Kurihara, Hiroki
中科院分区：
文献类型：
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作者：
Kawamura, Yumiko;Uchijima, Yasunobu;Kurihara, Hiroki
Sirtuins是一个系统发育保守的NAD（+）依赖性蛋白去乙酰化酶/ adp -核糖基转移酶家族，与多种生物过程有关。一些家族成员定位于线粒体，其功能被认为决定着床前胚胎的发育潜力。因此，我们在体外培养条件下描述了sirtuins在小鼠着床前发育中的作用。所有sirtuin成员均在卵子中表达，直到囊胚期表达逐渐减少。用sirtuin抑制剂治疗导致细胞内ROS水平升高，囊胚形成减少。这些影响通过sirna诱导的Sirt3（参与线粒体能量代谢）和Sirt3（-/-）胚胎的敲低得以再现。抗氧化剂n -乙酰- l-半胱氨酸和低氧条件挽救了这些不良反应。长期培养后将sirt3敲低胚胎移植到假孕小鼠体内，着床率和胎儿生长速率均下降，说明sirt3敲低胚胎对体外条件敏感，且效果持久。进一步的实验表明，母系衍生的Sirt3至关重要。Sirt3失活增加线粒体ROS产生，导致p53上调和下游基因表达改变。p53的失活改善了sirt3敲低胚胎的发育结果，表明ROS-p53通路是发育缺陷的原因。这些结果表明Sirt3在着床前胚胎在体外受精和培养过程中对应激条件起保护作用。
Sirtuins are a phylogenetically conserved NAD(+)-dependent protein deacetylase/ADP-ribosyltransferase family implicated in diverse biological processes. Several family members localize to mitochondria, the function of which is thought to determine the developmental potential of preimplantation embryos. We have therefore characterized the role of sirtuins in mouse preimplantation development under in vitro culture conditions. All sirtuin members were expressed in eggs, and their expression gradually decreased until the blastocyst stage. Treatment with sirtuin inhibitors resulted in increased intracellular ROS levels and decreased blastocyst formation. These effects were recapitulated by siRNA-induced knockdown of Sirt3, which is involved in mitochondrial energy metabolism, and in Sirt3(-/-) embryos. The antioxidant N-acetyl-L-cysteine and low-oxygen conditions rescued these adverse effects. When Sirt3-knockdown embryos were transferred to pseudopregnant mice after long-term culture, implantation and fetal growth rates were decreased, indicating that Sirt3-knockdown embryos were sensitive to in vitro conditions and that the effect was long lasting. Further experiments revealed that maternally derived Sirt3 was critical. Sirt3 inactivation increased mitochondrial ROS production, leading to p53 upregulation and changes in downstream gene expression. The inactivation of p53 improved the developmental outcome of Sirt3-knockdown embryos, indicating that the ROS-p53 pathway was responsible for the developmental defects. These results indicate that Sirt3 plays a protective role in preimplantation embryos against stress conditions during in vitro fertilization and culture.