STUDIES ON REGULATION OF DEVELOPMENT BY GENOMIC IMPRINTING

基因组印记发育调控的研究

• 批准号: 10660274
• 负责人: KONO Tomohiro
• 金额: $ 2.05万
• 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 (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660274/
• 关键词: GENOMIC IMPRINTING; MOUSE; DEVELOPMENT; NUCLEAR TRANSFER; GENE EXPRESSION; ゲノムインプリント; 胚発生; インプリンティング

This project was carried out to clarify when the genomic imprinting during oocyte growth is established in mice by investigations on imprinted gene expression and embryo development. Growing stage oocytes lack ability to mature into the metaphase of the second meiosis and support the development. To know nuclear competence of growing stage oocytes, fully-grown oocytes with nuclei from growing stage one were constructed by nuclear transfer. Following in vitro fertilization, the oocytes contained nuclei from growing one, 60μm in diameter, of juvenile mice were able to develop term. When oocytes from adult mice ovary were used as nuclear donors, the nucleus from oocytes of 50μm in diameter supported the term development. To evaluate the imprinted status in response to the developments, the expression of imprinted genes in the fetuses derived from constructed oocytes was examined by RT-PCR. The results showed that paternally and maternally expressed genes were activated and inhibited respectively in non-growing oocyte alleles, and that the regulation was established individually from early to late growing stage in oocytes. These results demonstrate that modifications in the maternal genome that occur during oocyte growth are essential for mammalin development.

该项目旨在通过研究印记基因表达和胚胎发育来阐明小鼠卵母细胞生长过程中的基因组印记何时建立。生长阶段的卵母细胞缺乏成熟到第二次减数分裂中期并支持发育的能力。为了了解生长阶段卵母细胞的核能力，通过核移植构建了具有生长第一阶段细胞核的完全生长的卵母细胞。体外受精后，卵母细胞中含有来自一只直径为 60 微米的幼年小鼠的细胞核，能够足月发育。当使用成年小鼠卵巢的卵母细胞作为核供体时，直径为50μm的卵母细胞的细胞核支持足月发育。为了评估印记状态对发育的反应，通过 RT-PCR 检查了来自构建卵母细胞的胎儿中印记基因的表达。结果表明，在非生长卵母细胞等位基因中，父本和母本表达的基因分别被激活和抑制，并且卵母细胞从早期到晚期的调节是单独建立的。这些结果表明，卵母细胞生长过程中发生的母体基因组修饰对于哺乳动物的发育至关重要。

项目成果

Kono, T.: "Influence of epigenetic changes during oocyte growth on nuclear reprogramming after nuclear tranfer."Journal of Fertility and Development. 10. 593-598 (1998)

Kono, T.：“卵母细胞生长过程中表观遗传变化对核移植后核重编程的影响。”生育与发育杂志。

Ohta, Y., Kono, T. et al: "Disruption of primary imprinting during oocyte growth leads to the modified expression of in printed genes during em bnyogenesis"Development. 125. 1553-1560 (1998)

Ohta, Y., Kono, T. 等人：“卵母细胞生长过程中初级印记的破坏导致胚胎发生过程中印记基因的表达发生改变”。

Kono T: "Influence, of epigenetic changes during oocytegrowth on nuclear reprogramming after nuclear transfer"Journal of Fertility and Development. 10. 593-598 (1998)

Kono T：“卵母细胞生长过程中表观遗传变化对核移植后核重编程的影响”《生育与发育杂志》。

Kono, T.: "The influence of epigenetic changes during oocyte growth on nuclear reprogramming after nuclear transfer"Journal of Fertility and Development. 10. 593-598 (1998)

Kono，T.：“卵母细胞生长过程中表观遗传变化对核移植后核重编程的影响”生育与发育杂志。

Obata, Y., Kaneko-Ishino, T., Koide, T., Takagi, Y., Ueda, T., Domeki, I.., Shiroishi, T., Ishino F. and Kono, T.: "Disruption of primary imprinting during oocyte growth leads to the modified expression of imprinted genes during embroygenesis."Development

Obata, Y.、Kaneko-Ishino, T.、Koide, T.、Takagi, Y.、Ueda, T.、Domeki, I..、Shiroishi, T.、Ishino F. 和 Kono, T.：“破坏