Spatial organization of cytoskeletal actin filaments and their regulatory proteins revealed by advanced methods in electron microscopy.

通过电子显微镜的先进方法揭示了细胞骨架肌动蛋白丝及其调节蛋白的空间组织。

• 批准号: 22370056
• 负责人: USUKURA Jiro
• 金额: $ 12.23万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22370056/
• 关键词: アクチン; 空間構造; 細胞骨格; 核膜; 電子顕微鏡; 中間径線維; 微小管; ストレス線維; アクチン線維; 細胞周期; ネスプリン; 生物物理; ミオシン; 制御タンパク質; 発生・分化

Current study aims to reveal the spatial organization of cytoskeletal actin filaments in the periphery of nuclear envelope. Cytoskeletal actin filaments in living cell have been investigated so far exclusively withfluorescent light microscopy. Therefore, observation was restricted on the stress fiber with relatively strong fluorescence and at ventral side of the cell. In order to detect real spatial structure of cytoskeleton, high voltage TEM (1000 KV), high resolution SEM and immuno-freeze etching technique were applied to unroofed whole cells. Nuclear envelope was supplied with many cytoskeleton including intermediate filaments, actin filaments and microtubules. In particular, intermediate filaments, vimentin, extending from nuclear pore like a rosette formed complicated meshwork and covered the total surface. Many actin filaments were attached to nuclear envelope as well. Since S1 decoration showed both pointed and barbed ends clearly in the surface of nucleus, there seemed to be the terminations and origins of actin filaments on the nuclear envelope. Unfortunately, however, we were not able to trace entire actin filaments on the nuclear envelope, because all actin filaments were associated or covered with vimentin filaments on the surface of nucleus. Under careful observation of many electron micrographs, many actin filaments appeared to extend from nuclear pores. Indeed, Nesprin 1, actin binding protein, was detected inthe periphery of nuclear pores. Actin filaments extending from nuclear envelope were bundled gradually to form stress fibers. These are completely new concept on nuclear related actin filaments.

目前的研究旨在揭示核膜周围细胞骨架肌动蛋白丝的空间组织。迄今为止，仅用荧光光学显微镜研究了活细胞中的细胞骨架肌动蛋白丝。因此，观察仅限于荧光较强且位于细胞腹侧的应力纤维。为了检测细胞骨架的真实空间结构，将高压TEM（1000 KV）、高分辨率SEM和免疫冷冻蚀刻技术应用于无顶全细胞。核膜上有许多细胞骨架，包括中间丝、肌动蛋白丝和微管。特别是中间丝，波形蛋白，像玫瑰花结一样从核孔延伸出来，形成复杂的网状结构并覆盖整个表面。许多肌动蛋白丝也附着在核膜上。由于S1修饰在细胞核表面清晰地显示出尖头和倒刺末端，因此核膜上似乎存在肌动蛋白丝的终止和起点。然而不幸的是，我们无法追踪核膜上的整个肌动蛋白丝，因为所有肌动蛋白丝都与细胞核表面的波形蛋白丝相关联或覆盖。在仔细观察许多电子显微照片时，许多肌动蛋白丝似乎从核孔中延伸出来。事实上，在核孔外围检测到了肌动蛋白结合蛋白 Nesprin 1。从核膜延伸出来的肌动蛋白丝逐渐成束形成应力纤维。这些是核相关肌动蛋白丝的全新概念。

项目成果

全截標本を用いた超高圧電子顕微鏡による細胞質細胞骨格の空間構造解析

使用超高压电子显微镜使用全切标本对细胞质细胞骨架进行空间结构分析

• 发表时间: 2010
• 作者: 臼倉治郎;平嶋涼子;南方志帆;荒井重勇
• 通讯作者: 荒井重勇

Spatial arrangement of cytoskeletal actin filaments with reference to the association with nuclear membrane

细胞骨架肌动蛋白丝的空间排列与核膜的关联

• 发表时间: 2011
• 作者: Usukura J;Minakata S
• 通讯作者: Minakata S

Spindle microtubules generate tension-dependent changes in the distribution of inner kinetochore proteins.

• DOI: 10.1083/jcb.201012050
• 发表时间: 2011-04-04
• 期刊: The Journal of cell biology
• 作者: Suzuki A;Hori T;Nishino T;Usukura J;Miyagi A;Morikawa K;Fukagawa T
• 通讯作者: Fukagawa T

Spatial Arrangement of Cytoskeletal Actin filaments and spatial specificity of actin binding proteins

细胞骨架肌动蛋白丝的空间排列和肌动蛋白结合蛋白的空间特异性

• 发表时间: 2010
• 作者: Usukura J;Yoshimura A;Minakata S
• 通讯作者: Minakata S

Hybrid liposomes affect cellular lipid constituents and caveolae structure

混合脂质体影响细胞脂质成分和细胞膜穴结构

• 发表时间: 2012
• 期刊: Bioorg Med Chem Lett
• 影响因子: 2.7
• 作者: Cao K;Tanaka K;Komizu Y;Tamiya-Koizumi K;Murate T;Ueoka R;Kyogashima M;Usukura J;Takahashi T;Suzuki M
• 通讯作者: Suzuki M