Epigenetic reprogramming and chromatin dynamics tin the course of mammalian oocyte development and fertilization

表观遗传重编程和染色质动力学影响哺乳动物卵母细胞发育和受精的过程

• 批准号: 14571584
• 负责人: TANAKA Mamoru
• 金额: $ 2.56万
• 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: 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-14571584/
• 关键词: Linker histone; Oocyte; FRAP; Chromatin remodeling; Clone; Reprogramming; 卵子; ゲノムリプログラミング

We tried to determine whether the replacement of somatic linker histones with H1foo occurs during the process of mouse nuclear transfer. H1foo was detected in the donor nucleus soon after transplantation. Thereafter, H1foo was restricted to the chromatin in up to two-cell stage embryos. After fusion of an oocyte with a cell expressing GFP (green fluorescent protein)-tagged somatic linker histone H1c, immediate release of H1c in the donor nucleus was observed. In addition, we found that H1foo is more mobile than H1c in living cells using fluorescence recovery after photobleaching (FRAP). The greater mobility of H1foo may contribute to its rapid replacement and the less stability of an embryonic chromatin structure. These results suggest that rapid replacement of H1c with H1foo may play a significant role in nuclear remodeling. Finally, we identified human H1foo using direct RT-nested PCR of a single human oocyte.Taken together, we conclude that the replacement of oocyte specific linker histones in the donor or sperm nucleus leads to the less stable assembly of chromatin, and thus it might contribute to the assembly of embryonic chromatin structures that are more easily replicated and transcribed when zygotic gene activation occurs.

我们试图确定在小鼠核移植过程中是否发生了用H1foo替换体细胞连接器组蛋白的情况。移植后即刻，供核内可检测到H1foo。此后，H1foo被限制在最多两个细胞期的胚胎中的染色质。卵母细胞与表达GFP(绿色荧光蛋白)标记的体细胞连接蛋白H1c的细胞融合后，观察到H1c在供体核中立即释放。此外，我们还利用光漂白后荧光恢复技术(FRAP)发现H1foo比H1c在活细胞中具有更强的流动性。H1foo的较大流动性可能有助于其快速替代，以及胚胎染色质结构的稳定性较差。这些结果表明，H1foo对H1c的快速替代可能在核重塑中发挥重要作用。最后，我们通过对单个人卵母细胞的直接RT-巢式聚合酶链式反应鉴定了人H1foo。综上所述，我们得出结论，卵母细胞特异性连接器组蛋白在供体或精子核中的替换导致染色质组装不太稳定，因此它可能有助于胚胎染色质结构的组装，当合子基因激活时，这些结构更容易复制和转录。

项目成果

Tanaka M, et al.: "The generation and characterization of an ovary-selective cDNA library."Molecular Cellular Endocrinology. 202 1. 67-69 (2003)

Tanaka M 等人：“卵巢选择性 cDNA 文库的生成和表征。”分子细胞内分泌学。

Tanaka M, et al.: "H1oo : a pre-embryonic H1 linker histone in search of a function."Molecular Cellular Endocrinology. 202 1. 5-9 (2003)

Tanaka M 等人：“H1oo：寻找功能的胚胎前 H1 连接组蛋白。”分子细胞内分泌学。

Tanaka M, et al.: "The generation and characterization of an ovary-selective cDNA library."Molecular Cellular Endocrinology. 202. 67-69 (2003)

Tanaka M 等人：“卵巢选择性 cDNA 文库的生成和表征。”分子细胞内分泌学。

Tanaka m, Teranishi T, Miyakoshi K, Jiaxue W, Ueno K, Matsumoto T, Hattori Y, Yoshimura Y.: "Oocyte-specific linker histone in mammalian species."J Mamm Ova Res.. 19. 61-65 (2002)

Tanaka m、Teranishi T、Miyakoshi K、Jiaxue W、Ueno K、Matsumoto T、Hattori Y、Yoshimura Y.：“哺乳动物物种中的卵母细胞特异性连接组蛋白。”J Mamm Ova Res.. 19. 61-65 (2002)

Tanaka Y, et al.: "Structure and expression of the human oocyte-specific histone H1 gene elucidated by direct RT-nested PCR of a single oocyte."Biochem Biophys Res Commun. 304. 351-357 (2003)

Tanaka Y 等人：“通过单个卵母细胞的直接 RT 巢式 PCR 阐明了人类卵母细胞特异性组蛋白 H1 基因的结构和表达。”Biochem Biophys Res Commun。