STED super-resolution imaging of mitochondria labeled with TMRM in living cells
STED super-resolution imaging of mitochondria labeled with TMRM in living cells
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DOI：
10.1016/j.mito.2016.03.009
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发表时间：
2016-05-01
期刊：
影响因子：
4.4
通讯作者：
Ohsawa, Ikuroh
中科院分区：
文献类型：
--
作者：
Ishigaki, Masaya;Iketani, Masumi;Ohsawa, Ikuroh
我们应用受激发射损耗（STED）成像与亚衍射分辨率的线粒体亚线粒体结构。它们的形状取决于细胞的类型和生理状态。用阳离子荧光染料四甲基罗丹明甲酯（TMRM）染色，揭示了层状结构的有趣细节，包括快速变化的幕状结构。TMRM阳性结构既不与基质中的蛋白质共定位，也不与外膜上的蛋白质共定位，但部分与类核共定位。抑制线粒体接触部位的成分破坏了板层TMRM阳性结构。氧化磷酸化系统的解偶联从内膜释放TMRM，而基质结构没有任何改变。STED图像进一步表明，电子传递链的复合物位于TMRM阳性结构的表面上。这里提出的方法提供了新的见解在体内性质的submitochondrial结构，并可用于进一步的功能调查，这些复杂的结构。(C)2016 Elsevier B.V.和线粒体研究学会。All rights reserved.
We applied stimulated emission depletion (STED) imaging with subdiffraction resolution to submitochondrial structures in mitochondria. Their shapes depend on both a cell's type and its physiological state. Staining with a cationic fluorescent dye, tetramethylrhodamine methyl ester (TMRM), unveiled intriguing details of lamellar structure, consisting of rapidly changeable, curtain-like formations. The TMRM-positive structure colocalized with neither proteins in the matrix nor on the outer membrane, but partially localized with the nucleoid. Suppression of a component in the mitochondrial contact site disrupted the lamellar TMRM-positive structure. Uncoupling of the oxidative phosphorylation system released TMRM from the inner membrane without any alteration in the matrix structure. STED images further showed that complexes of the electron transport chain are located on the surface of TMRM-positive structures. The approach presented here provides novel insights into the in vivo nature of submitochondrial structures, and can be used for further functional investigations of these complex structures. (C) 2016 Elsevier B.V. and Mitochondria Research Society. All rights reserved.