Melatonin Enhances the Development of Porcine Cloned Embryos by Improving DNA Methylation Reprogramming
Melatonin Enhances the Development of Porcine Cloned Embryos by Improving DNA Methylation Reprogramming
复制标题
褪黑素通过改善 DNA 甲基化重编程促进猪克隆胚胎的发育
DOI：
10.1089/cell.2019.0103
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发表时间：
2020-03-24
影响因子：
1.6
通讯作者：
Huan, Yanjun
中科院分区：
文献类型：
--
作者：
Qu, Jiadan;Sun, Mingjun;Huan, Yanjun
克隆胚胎中不完全的DNA甲基化重编程导致克隆效率低下。褪黑激素已被证明可以促进克隆胚胎的发育，但褪黑激素在体细胞核移植中的作用尚不清楚。本研究表明，10（-7）M褪黑素能显著促进克隆胚胎的发育进程，降低合子基因组激活前的阻滞率，并能上调克隆胚胎的囊胚率。褪黑激素还通过上调抗凋亡因子的表达同时下调促凋亡基因的转录，促进假原核的形成，增加囊胚细胞数量，减少胚胎凋亡。进一步的研究表明，褪黑素处理后，克隆胚胎中DNA甲基化重编程相关基因的表达显著提高;褪黑素有效促进了克隆胚胎基因组DNA去甲基化和DNA重甲基化、多能性相关基因Oct 4的DNA去甲基化、印迹基因H19/Igf 2的DNA甲基化维持和组织特异性基因Thy 1的DNA重甲基化。因此，合子基因组激活相关基因Eif 1a、多能性相关基因Oct 4、Nanog和Sox 2、印迹基因Igf 2和H19以及囊胚质量相关基因Cdx 2和ATP 1b 1显著上调，组织特异性基因Thy 1和Col 5a 2显著沉默。总之，褪黑激素通过改善DNA甲基化重编程来促进克隆胚胎的发育。这项工作揭示了褪黑激素可以调节DNA甲基化重编程，并提供了一个新的见解，以提高克隆效率。
Incomplete DNA methylation reprogramming in cloned embryos leads to poor cloning efficiency. Melatonin has been proven to improve the development of cloned embryos, however, the role of melatonin during somatic cell nuclear transfer remains unclear. This work demonstrated that 10(-7) M melatonin significantly enhanced the developmental progress, reduced the arrested rate before zygotic genome activation, and upregulated the blastocyst rate of cloned embryos. Melatonin also promoted the pseudo-pronucleus formation, increased blastocyst cell number, and reduced embryo apoptosis through upregulating the expression of antiapoptosis factors while downregulating the transcription of proapoptosis genes. Further study displayed that DNA methylation reprogramming related genes were greatly improved in cloned embryos when treated with melatonin; then, melatonin effectively promoted genomic DNA demethylation and DNA remethylation, DNA demethylation of pluripotency related gene Oct4, DNA methylation maintenance of imprinted gene H19/Igf2, and DNA remethylation of tissue-specific gene Thy1 in cloned embryos. Thus, zygotic genome activation related gene Eif1a, pluripotency related genes Oct4, Nanog, and Sox2, imprinted genes Igf2 and H19, and blastocyst quality related genes Cdx2 and ATP1b1 were remarkably upregulated, and tissue-specific genes Thy1 and Col5a2 were considerably silenced. In conclusion, melatonin enhanced the development of cloned embryos by ameliorating DNA methylation reprogramming. This work reveals that melatonin can regulate DNA methylation reprogramming and provides a novel insight to improve cloning efficiency.