The cytoplasm of mouse germinal vesicle stage oocytes can enhance somatic cell nuclear reprogramming
The cytoplasm of mouse germinal vesicle stage oocytes can enhance somatic cell nuclear reprogramming
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DOI：
10.1242/dev.023747
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发表时间：
2008-12-01
期刊：
影响因子：
4.6
通讯作者：
Wakayama, Teruhiko
中科院分区：
文献类型：
--
作者：
Bui, Hong-Thuy;Wakayama, Sayaka;Wakayama, Teruhiko
在哺乳动物克隆中，有证据表明基因组重编程因子位于卵母细胞或受精卵的细胞核而不是细胞质中。然而，人们对重编程的机制知之甚少，使用核因子的新方法也没有成功地从分化的体细胞核中产生克隆小鼠。我们的目的是确定是否有功能性重编程因子存在于细胞质中的萌发囊泡阶段（GV）的卵母细胞。我们发现，GV卵母细胞的细胞质可以重塑体细胞核，完全脱甲基组蛋白H3的赖氨酸9和部分脱乙酰组蛋白H3的赖氨酸9和14。此外，GV卵母细胞的细胞质裂解物促进体细胞重编程和克隆胚胎发育，通过测量组蛋白H3-K9低甲基化，Oct 4和Cdx 2在囊胚中的表达，以及克隆后代的生产进行评估。因此，基因组重编程因子存在于GV卵母细胞的细胞质中，并且可以促进克隆技术。这一发现也有助于研究组蛋白去乙酰化和去甲基化的机制，尽管组蛋白甲基化被认为是表观遗传学稳定的。
In mammalian cloning, evidence suggests that genomic reprogramming factors are located in the nucleus rather than the cytoplasm of oocytes or zygotes. However, little is known about the mechanisms of reprogramming, and new methods using nuclear factors have not succeeded in producing cloned mice from differentiated somatic cell nuclei. We aimed to determine whether there are functional reprogramming factors present in the cytoplasm of germinal vesicle stage (GV) oocytes. We found that the GV oocyte cytoplasm could remodel somatic cell nuclei, completely demethylate histone H3 at lysine 9 and partially deacetylate histone H3 at lysines 9 and 14. Moreover, cytoplasmic lysates of GV oocytes promoted somatic cell reprogramming and cloned embryo development, when assessed by measuring histone H3-K9 hypomethylation, Oct4 and Cdx2 expression in blastocysts, and the production of cloned offspring. Thus, genomic reprogramming factors are present in the cytoplasm of the GV oocyte and could facilitate cloning technology. This finding is also useful for research on the mechanisms involved in histone deacetylation and demethylation, even though histone methylation is thought to be epigenetically stable.