Imprinting disorder in donor cells is detrimental to the development of cloned embryos in pigs.
Imprinting disorder in donor cells is detrimental to the development of cloned embryos in pigs.
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供体细胞的印迹紊乱不利于猪克隆胚胎的发育
DOI：
10.18632/oncotarget.20390
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发表时间：
2017-09-22
期刊：
影响因子：
--
通讯作者：
Huan Y
中科院分区：
文献类型：
--
作者：
Song X;Li F;Jiang Z;Sun Y;Li H;Gao S;Zhang L;Xue B;Zhao G;Li J;Liu Z;He H;Huan Y
体细胞核移植过程中的印记障碍通常会导致克隆动物的异常和克隆效率低下。然而，关于供体细胞印迹在克隆胚胎发育中的作用知之甚少。在这里，我们证明了（H19/Igf 2）在形态学异常克隆胎儿来源的猪胎儿成纤维细胞中的表达（异常印迹组）的甲基化程度更低，因此，与形态学正常的克隆胎儿来源的成纤维细胞相比，H19转录显著升高，Igf 2表达显著降低（正常印迹组）或供体胎儿成纤维细胞（对照组）。当这些成纤维细胞被用作供体细胞时，异常印迹组显示出更低的印迹甲基化水平，对应于Dnmt 1，Dnmt 3a和Zfp 57的表达显著下调，以及显著降低的囊胚率，而正常印迹组呈现与对照组相似的印迹模式，基因表达和胚胎发育。当5-aza-dC降低正常印迹组成纤维细胞印迹甲基化水平时，克隆胚胎显示更严重的印迹受损和显着降低的囊胚率。当印迹异常组中上调的H19转录被敲除时，印迹状态被部分挽救，克隆胚胎的卵裂率和囊胚率显著提高。总之，供体细胞印迹障碍降低了克隆胚胎的发育效率。本研究为深入了解供体细胞调控克隆胚胎发育的分子机制提供了新的思路。
Imprinting disorder during somatic cell nuclear transfer usually leads to the abnormality of cloned animals and low cloning efficiency. However, little is known about the role of donor cell imprinting in the development of cloned embryos. Here, we demonstrated that the imprinting (H19/Igf2) in porcine fetus fibroblasts derived from the morphologically abnormal cloned fetuses (the abnormal imprinting group) was more hypomethylated, and accordingly, significantly higher H19 transcription and lower Igf2 expression occurred in comparison with those in fibroblasts derived from morphologically normal cloned fetuses (the normal imprinting group) or donor fetus fibroblasts (the control group). When these fibroblasts were used as donor cells, the abnormal imprinting group displayed an even lower imprinting methylation level, in correspondence to the significantly downregulated expression of Dnmt1, Dnmt3a and Zfp57, and a markedly reduced blastocyst rate, while the normal imprinting group took on the similar patterns of imprinting, gene expression and embryo development to the control group. When 5-aza-dC was applied to reduce the fibroblasts imprinting methylation level in the normal imprinting group, cloned embryos displayed the more severely impaired imprinting and significantly lower blastocyst rate. While the upregulated H19 transcription in the abnormal imprinting group was knocked down, the imprinting statuses were partly rescued, and the cleavage and blastocyst rates significantly increased in cloned embryos. In all, donor cell imprinting disorder reduced the developmental efficiency of cloned embryos. This work provides a new insight into understanding the molecular mechanism of donor cells regulating the cloned embryo development.
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DOI：
10.1262/jrd.2015-048
发表时间：
2016
期刊：
The Journal of reproduction and development
影响因子：
--
作者：
Huan YJ;Wu ZF;Zhang JG;Zhu J;Xie BT;Wang JY;Li JY;Xue BH;Kong QR;Liu ZH
通讯作者：
Liu ZH
影响因子：
3.7
作者：
Xu W;Li Z;Yu B;He X;Shi J;Zhou R;Liu D;Wu Z
通讯作者：
Wu Z
DNA甲基化重编程相关基因的表达模式与猪合子基因组激活后克隆胚胎的发育能力相关
影响因子：
1.2
作者：
Huan, Yanjun;Wang, Hongmei;He, Hongbin
通讯作者：
He, Hongbin
影响因子：
1.6
作者：
Shen, Chih-Jie;Lin, Chiao-Chieh;Chen, Chuan-Mu
通讯作者：
Chen, Chuan-Mu
影响因子：
7.2
作者：
Barlow, Denise P.;Bartolomei, Marisa S.
通讯作者：
Bartolomei, Marisa S.