Epigenetics and Chromatin Remodeling in mammalian early development

哺乳动物早期发育中的表观遗传学和染色质重塑

基本信息

批准号：
16591684
负责人：
TANAKA Mamoru
金额：
$ 2.3万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

Reverse genetic approaches have become increasingly attractive in developmental biology. Recently, one strategy for inhibiting gene function has been shown to be extremely effective. Morpholino antisense oligonucleotides (morpholinos) block translation by preventing ribosomes from binding to target mRNA. The use of morpholino is proving to be a valuable tool for investigating gene function in zebrafish and xenopus embryos. However, few studies have evaluated the microinjection of morpholinos into mammalian oocytes. The mouse oocyte-specific linker histone H1foo is thought to play a key role in the control of gene expression and chromatin modification during oogenesis and early embryogenesis. To investigate the feasibility of using morpholinos to suppress mouse gene function, we performed the microinjection of morpholinos against H1foo into mouse GV stage oocytes. Immunoblotting and immunostaining analysis showed that the exression of H1foo protein was successfully suppressed in the oocytes injected with H1foo morpholinos, and in H1foo morpholinos injected group, the rate of in vitro maturation was lower than that in control group. This result suggests that H1foo is essential for maturation of GV-stage oocytes. Morpholinos provide a useful method to study gene function also in the mouse development.

反向遗传方法在发育生物学中变得越来越有吸引力。最近，一种抑制基因功能的策略已被证明非常有效。吗啉代反义寡核苷酸（吗啉代）通过阻止核糖体与靶 mRNA 结合来阻断翻译。事实证明，吗啉代的使用是研究斑马鱼和爪蟾胚胎基因功能的宝贵工具。然而，很少有研究评估吗啉代显微注射到哺乳动物卵母细胞中。小鼠卵母细胞特异性接头组蛋白 H1foo 被认为在卵子发生和早期胚胎发生过程中基因表达和染色质修饰的控制中发挥关键作用。为了研究使用吗啉代抑制小鼠基因功能的可行性，我们将针对 H1foo 的吗啉代显微注射到小鼠 GV 期卵母细胞中。免疫印迹和免疫染色分析表明，注射H1foo吗啉代的卵母细胞中H1foo蛋白的表达被成功抑制，且注射H1foo吗啉组的体外成熟率低于对照组。这一结果表明 H1foo 对于 GV 期卵母细胞的成熟至关重要。吗啡啉提供了一种有用的方法来研究小鼠发育中的基因功能。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Chromatin remodeling with oocyte-specific linker histones

使用卵母细胞特异性连接组蛋白进行染色质重塑

DOI：
发表时间：
2004
期刊：
J Mamm Ova Res 21
影响因子：
0
作者：
Tanaka M;Teranishi T;Yoshimura Y
通讯作者：
Yoshimura Y

Rapid replacement of somatic linker histones with the oocyte-specific linker histone H1foo in nuclear transfer.

DOI：
10.1016/j.ydbio.2003.10.004
发表时间：
2004-02
期刊：
Developmental biology
影响因子：
2.7
作者：
Takahide Teranishi;Mamoru Tanaka;Shingo Kimoto;Y. Ono;K. Miyakoshi;T. Kono;Y. Yoshimura
通讯作者：
Takahide Teranishi;Mamoru Tanaka;Shingo Kimoto;Y. Ono;K. Miyakoshi;T. Kono;Y. Yoshimura

Rapid replacement of somatic linker histones with the oocyte-specific linker histone Hlfoo in nuclear transfer.

在核移植中用卵母细胞特异性连接组蛋白 Hlfoo 快速替换体细胞连接组蛋白。