Imaging analysis of stress signaling and spatio-temporal dynamics of MAPKKK activity

MAPKKK 活性的应激信号和时空动态的成像分析

• 批准号: 21770137
• 负责人: TOMIDA Taichiro
• 金额: $ 2.91万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21770137/
• 关键词: ストレス応答; キナーゼ; イメージング; ストレス; FRET

(1) Automated FRET analysis system was established by which intracellular MAPK activity from more than 1,000 cells can be imaged simultaneously in real-time. (2) By administrating MAPK activity FRET reporter to subcellular components, spatial distribution of stress-activated MAPK activity was determined. I found that ribotoxic stress, UV, hyperosmotic stress, and interleukin-1 induces p38 MAPK activation in cytoplasm. (3) Before this study, individual cellular response in mammalian cells has been not well understood because of the lack of the tools to determine individual cellular response with a good quantitativity. Using automated FRET system and MAPK activity indicator, I identified that ribotoxic stress response in individual cell was similar to average response of cell population at any doses applied.

(1)建立了FRET自动分析系统，可同时对1000多个细胞的MAPK活性进行实时检测。(2)用MAPK活性FRET报告基因定位亚细胞成分，确定应激激活的MAPK活性的空间分布。我发现核毒应激、紫外线、高渗应激和白介素1可诱导细胞质中p38MAPK的激活。(3)在这项研究之前，由于缺乏能够很好地定量确定单个细胞反应的工具，对哺乳动物细胞中的个体细胞反应还没有很好的了解。使用自动FRET系统和MAPK活性指示器，我发现在任何剂量下，单个细胞的核毒应激反应与细胞群体的平均反应相似。

项目成果

Stimulus-specific distinctions in spatial and temporal dynamics of stress-activated protein kinase kinase kinases revealed by a fluorescence resonance energy transfer biosensor.

荧光共振能量转移生物传感器揭示了应激激活蛋白激酶激酶激酶在空间和时间动力学上的刺激特异性差异。

• 发表时间: 2009
• 期刊: Molecular and Cellular Biology Vol.29、No.22
• 作者: Taichiro Tomida;Pauline O'Grady;Mutsuhiro Takekawa;Haruo Saito.
• 通讯作者: Haruo Saito.

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Stimulus-Specific Distinctions in Spatial and Temporal Dynamics of Stress-Activated Protein Kinase Kinase Kinases Revealed by a Fluorescence Resonance Energy Transfer Biosensor

荧光共振能量转移生物传感器揭示了应激激活蛋白激酶的空间和时间动力学的刺激特异性差异

• 发表时间: 2009
• 期刊: Molecular and Cellular Biology 29
• 作者: Taichiro Tomida;Mutsuhiro Takekawa;Pauline O'Grady;Haruo Saito
• 通讯作者: Haruo Saito

線虫感覚神経のMAP キナーゼリン酸化シグナルのin vivoイメージング解析

线虫感觉神经元 MAP 激酶磷酸化信号的体内成像分析

• 发表时间: 2011
• 作者: 冨田太一郎;斎藤春雄
• 通讯作者: 斎藤春雄

線虫感覚神経のMAPキナーゼリン酸化シグナルのin vivoイメージング解析

线虫感觉神经元 MAP 激酶磷酸化信号的体内成像分析

• 发表时间: 2011
• 作者: 冨田太一郎;斎藤春雄
• 通讯作者: 斎藤春雄