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Control of epigenetic modification for oocytes genome

卵母细胞基因组表观遗传修饰的控制

基本信息

批准号：
14360170
负责人：
KONO Tomohiro
金额：
$ 9.6万
依托单位：
TOKYO UNIVERSITY OF AGRICULTURE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14360170/
关键词：
Oocytes Nuclear transfer Genomic imprinting In vitro culture Development 体外培養

项目摘要

In this study we evaluated epigenetic modification during oocyte growth in mice and pig in relation to in vitro production of oocytes. The results obtained were summarized as follows. 1)Production system for competent oocytes commencing from day 12.5 fetal ovaries has been developed in vitro. The fetal ovary with mesonephroi was cultured in vitro for 8 days, and followed by 10 days culture after removal of mesonephroi. Developed follicles were dissected and collected from the ovaries, and then the follicles were further cultured for 11 days. Finally, oocytes developed over 60μm in diameter were obtained. 2)To evaluate epigenetic status, methylation status of the differentially methylated region of Igf2r, Zac1 and H19 genes were analyzed in the in vitro produced oocytes. The results showed that oocytes acquired appropriate epigenetic modification during their growth period in vitro. Nuclear transfer study revealed that the genome derived from in vitro produced oocytes was competent to support normal development. 3)In pig, significantly small oocytes, which were collected from adult ovary, were applied for in vitro culture and the full size oocytes were successfully obtained. 4)Molecular analysis for parental imprinting, we detected that 27 of imprinted genes are epigenetically modified in the female germ line. Together, the present study show that genomic imprinting in female germ line plays definitive role for supporting normal development.

在这项研究中，我们评估了小鼠和猪卵母细胞生长过程中的表观遗传修饰与卵母细胞体外生产的关系。所得结果总结如下。1)建立了从12.5天胎儿卵巢开始的体外成熟卵母细胞生产系统。将带中肾的胎儿卵巢体外培养8天，去除中肾后继续培养10天。从卵巢中解剖并收集已发育的卵泡，然后将卵泡进一步培养11天。最后获得直径大于60μm的卵母细胞。2）分析体外培养卵母细胞Igf2r、Zac1和H19基因甲基化差异区域的甲基化状态，评价表观遗传状态。结果表明，卵母细胞在体外发育过程中获得了适当的表观遗传修饰。核移植研究表明，来自体外产生的卵母细胞的基因组能够支持正常发育。3）从成年猪卵巢中采集体积较小的卵母细胞进行体外培养，获得了完整大小的卵母细胞。4)通过对亲本印迹的分子分析，我们检测到27个印迹基因在雌性生殖系中被表观遗传修饰。总之，目前的研究表明，基因组印记在女性生殖系中起着决定性的作用，以支持正常的发展。