Mechanism for the acquisition of totipotency by Yamanaka factors

山中因子获得全能性的机制

• 批准号: 22658091
• 负责人: AOKI Fugaku
• 金额: $ 2.32万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22658091/
• 关键词: 卵; 初期胚; リプログラミング; iPS細胞; 山中因子; 受精卵; 全能性

The expression of the transcripts of Yamanaka factors was examined by RT-PCR, which revealed that all of the factors were detected in the preimplantation embryos. Since the expression levels of Oct3/4 and c-Myc seemed to be high, I focused on these genes in the further experiments. Immunocytochemistry revealed that the nuclear localization of both of OCT3/4 and c-MYC proteins increased after fertilization. When full-grown oocytes were microinjected with Oct3/4 siRNA and then subjected to in vitro fertilization, the fertilized oocytes did not cleave to the 2-cell stage. In these oocytes, the expressions of the genes which are usually expressed after fertilization were reduced.

通过RT-PCR检查Yamanaka因子转录本的表达，结果表明在植入前胚胎中检测到所有因子。由于Oct3/4和c-Myc的表达水平似乎很高，因此我在进一步的实验中重点关注这些基因。免疫细胞化学显示受精后 OCT3/4 和 c-MYC 蛋白的核定位均增加。当对完全生长的卵母细胞显微注射Oct3/4 siRNA，然后进行体外受精时，受精的卵母细胞并未分裂至2细胞阶段。在这些卵母细胞中，通常在受精后表达的基因的表达减少。

项目成果

受精前後におけるゲノムリプログラミングへのヒストン変異体置換の関与

组蛋白变异替换参与受精前后基因组重编程

• 发表时间: 2012
• 作者: 青木不学