多能性幹細胞の分化過程における染色体の機能的構造変化

多能干细胞分化过程中染色体功能结构的变化

• 批准号: 12F02079
• 负责人: 中辻 憲夫
• 金额: $ 1.34万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12F02079/
• 关键词: Stem cells; Histones; Acetylation; Chromatin; Trophoblast; Differentiation; Pluripotency; Immunofluorescence; 超解像度顕微鏡; ヒトES細胞; ヒストン修飾; ヒストンバリアント; 細胞分化; エピジェネティクス; クロマチン

My research project investigated the cellular events involved in the differentiation of human embryonic stem (hES) cells into a specialized type of cell called a trophoblast cell. The major part of the project was centered on understanding how chromatin (the architecture of the nucleus) changes when hES cells loose their pluripotency and differentiate into the specialized trophoblast cells. I specifically studied proteins called 'histones' which are the basic unit of chromatin. I was particularly interested in a type of histone protein called 'histone 2A variant Z' (H2AZ). I was interested in H2AZ because it can have different effects on the chromatin architecture when by becoming modified. These modifications primarily concern the addition of small molecules called "acetylation" or "methylation". To study the changes in the modification of H2AZ I used a technique called immunofluorencese (to fluorescently label the proteins or RNA molecules I want to study) I then imaged the cell nucleus with a very sensitive microscope and then perform a specialized type of image processing and analysis that was writen in my laboratory on the captured image. Using these techniques, I found that status of H2AZ acetylation in the stem cell changes very quickly (in as little as 12 hours) following treatment to induce differentiation. I have studied the protein complexes that both acetylate and de-acetylate H2AZ under hES differentiation and found that de-acetylation of H2AZ is critical for differentiation to proceed.

我的研究项目调查了人类胚胎干 (hES) 细胞分化为一种称为滋养层细胞的特殊类型细胞所涉及的细胞事件。该项目的主要部分集中于了解当 hES 细胞失去多能性并分化为专门的滋养层细胞时染色质（细胞核的结构）如何变化。我专门研究了称为“组蛋白”的蛋白质，它是染色质的基本单位。我对一种称为“组蛋白 2A 变体 Z”(H2AZ) 的组蛋白特别感兴趣。我对 H2AZ 很感兴趣，因为它经过修饰后会对染色质结构产生不同的影响。这些修饰主要涉及添加称为“乙酰化”或“甲基化”的小分子。为了研究 H2AZ 修饰的变化，我使用了一种称为免疫荧光的技术（荧光标记我想要研究的蛋白质或 RNA 分子），然后用非常灵敏的显微镜对细胞核进行成像，然后执行专门类型的图像处理和我的实验室对捕获的图像进行的分析。使用这些技术，我发现在诱导分化的治疗后，干细胞中 H2AZ 乙酰化的状态变化非常快（只需 12 小时）。我研究了 hES 分化过程中乙酰化和去乙酰化 H2AZ 的蛋白质复合物，发现 H2AZ 去乙酰化对于分化的进行至关重要。

项目成果

Peripheral H2AZ. ac marks differentiating hES

外围 H2AZ。

• 发表时间: 2013
• 作者: Xiang-ning Meng;Wei-ling Wang;Miao-yong Zhu;R.O. Suzuki;Georgia R. Kafer
• 通讯作者: Georgia R. Kafer

The distribution of histone variant H2AFZ is restricted according to lineage commitment status in mouse trophohlast cells

组蛋白变体 H2AFZ 的分布根据小鼠滋养层细胞中的谱系定型状态而受到限制

• 发表时间: 2013
• 作者: Xiang-ning Meng;Wei-ling Wang;Miao-yong Zhu;R.O. Suzuki;Georgia R. Kafer;Georgia R. Kafer ; Peter L. Kaye ; Marie Pantaleon ; Peter M.
• 通讯作者: Georgia R. Kafer ; Peter L. Kaye ; Marie Pantaleon ; Peter M.