小鼠孤雌胚、体外培养胚与体内胚H3K9乙酰式的比较

doi:10.16288/j.yczz.2015.01.011

遗传 ›› 2015, Vol. 37 ›› Issue (1): 77-83.doi: 10.16288/j.yczz.2015.01.011

小鼠孤雌胚、体外培养胚与体内胚H3K9乙酰式的比较

陈利¹, 丁芳¹, 刘勇^{2, 3}, 吴风瑞^{2, 3}, 丁彪^{2, 3}, 王荣^{2, 3}, 李文雍²

1. 安徽大学生命科学学院,合肥 230039;
2. 胚胎发育与生殖调节安徽省重点实验室,阜阳 236041;
3. 阜阳师范学院生物与食品工程学院,阜阳 236041

收稿日期:2014-06-18 修回日期:2014-10-01 出版日期:2015-01-20 发布日期:2015-01-20
通讯作者: 李文雍,教授,硕士生导师,研究方向：胚胎发育分子生物学。E-mail: liwenyong@aliyun.com E-mail:liwenyong@aliyun.com
作者简介:陈利,硕士研究生,专业方向：发育分子生物学。E-mail：chenli3096@163.com
基金资助:
国家自然科学基金项目(编号：31372273,31201789),安徽省省级学科建设重大项目(编号：皖教秘科[2014]28号),安徽大学学术创新研究项目(编号：yqh100130),安徽省教育厅自然科学基金项目(编号：KJ2013A202)和安徽省自然科学基金项目(编号：1408085MC44, 1408085QC65)资助

Comparative analysis of H3K9 acetylation level in parthenogenetic, and in vitro and in vivo developed mouse embryos

Li Chen¹, Fang Ding¹, Yong Liu^{2, 3}, Fengrui Wu^{2, 3}, Biao Ding^{2, 3}, Rong Wang^{2, 3}, Wenyong Li²

1. College of Life Science, Anhui University, Hefei 230039, China; 2. Key Laboratory of Embryo Development and Reproductive Regulation of Anhui Province, Fuyang 236041, China; 3. College of Life Science, Fuyang Normal University, Fuyang 236041, China

Received:2014-06-18 Revised:2014-10-01 Online:2015-01-20 Published:2015-01-20

摘要/Abstract

摘要： 孤雌胚胎的发育率比体内体外生成胚胎的发育率要慢,为研究小鼠孤雌胚、体外培养胚H3K9乙酰化(H3K9ac)模式与体内自然胚之间的差异、曲古抑菌素A(Trichostatin,TSA)对孤雌胚H3K9乙酰化模式的影响及表观遗传模式对孤雌胚、体外培养胚发育的影响,文章采用间接免疫荧光法对小鼠植入前各时期孤雌胚、体外培养胚及体内自然胚基因组组蛋白的H3K9乙酰化水平进行检测。结果显示,植入前各时期孤雌胚H3K9乙酰化模式与体内组变化趋势基本一致,但平均荧光强度较体内组普遍偏高;经TSA处理后孤雌胚H3K9乙酰化水平有所提高,原核期至8-细胞期差异显著(P<0.05)。体外培养胚H3K9乙酰化荧光强度与体内组变化趋势也基本一致,但平均荧光强度较体内组普遍偏低。以上结果表明,小鼠孤雌胚H3K9乙酰化水平高于体内胚,使植入前胚胎发育过程中本应沉默的基因启动子发生超乙酰化,进而抑制胚胎发育,这可能是造成孤雌胚胎发育能力较差的重要原因之一;TSA处理可以部分弥补体外培养环境对胚胎发育带来的伤害,但TSA提高孤雌胚的发育能力可能并不完全是通过改变H3K9乙酰化水平来实现的。

关键词: 表观遗传修饰, 孤雌胚胎, H3K9乙酰化, 曲古抑菌素A, 小鼠

Abstract: The developmental rate of parthenogenetic embryos is slower than that of embryos generated in vitro and in vivo. To detect the effects of epigenetic modification on embryo development, we compared the H3K9 acetylation level in these three types of embryos as well as parthenogenetic embryos treated with a histone deacetylase inhibitor trichostatin (TSA) by indirect immunofluorescence. Our results showed that fluctuations in the level of acetylated H3K9 detected during embryo development are similar among different types of mouse embryos. However, the level of H3K9 acetylation in parthenogenetic embryos is significantly higher while the level in embryos generated in vitro is lower when compared with that in embryos derived from in vivo. Treatment of parthenogenetic embryos with TSA increases the developmental rate but further elevates the level of H3K9 acetylation, especially from pronuclear to 8-cell stages. These results suggest that the promoters of genes that should be silenced during pre-implantation embryo development may be hyperacetylated in parthenogenetic embryos which inhibit normal embryo development. However, the positive effect of TSA on embryo development is not through altering the H3K9 acetylation level.

Key words: epigenetic modification, parthenogenetic embryos, H3K9 acetylation, trichostatin A (TSA), mouse

陈利, 丁芳, 刘勇, 吴风瑞, 丁彪, 王荣, 李文雍. 小鼠孤雌胚、体外培养胚与体内胚H3K9乙酰式的比较[J]. 遗传, 2015, 37(1): 77-83.

Li Chen, Fang Ding, Yong Liu, Fengrui Wu, Biao Ding, Rong Wang, Wenyong Li. Comparative analysis of H3K9 acetylation level in parthenogenetic, and in vitro and in vivo developed mouse embryos[J]. HEREDITAS(Beijing), 2015, 37(1): 77-83.

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小鼠孤雌胚、体外培养胚与体内胚H3K9乙酰式的比较

Comparative analysis of H3K9 acetylation level in parthenogenetic, and in vitro and in vivo developed mouse embryos

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