Visualization of mitochondria in living cells with a genetically encoded yellow fluorescent protein originating from a yellow-emitting luminous bacterium.
Visualization of mitochondria in living cells with a genetically encoded yellow fluorescent protein originating from a yellow-emitting luminous bacterium.
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使用源自黄色发光细菌的基因编码黄色荧光蛋白对活细胞中的线粒体进行可视化。
DOI：
10.1039/c3pp25360k
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
(他8 名、8 番目)
中科院分区：
文献类型：
--
作者：
Karatani H;(他8 名、8 番目)
我们利用遗传编码的黄色荧光蛋白（Y1-Yellow）和常规荧光显微镜观察真核细胞模型酵母酵母（saccharomyces cerevisiaea）线粒体的氧化还原和结构变化。Y1-Yellow源自一种黄色发光细菌aliivibrio sifiaeY1，与线粒体靶向序列（mt-sequence）融合。y1 -黄色荧光仅在线粒体位点产生，黄色荧光的颜色很容易与细胞自身荧光和常规探针的荧光区分。Y1-Yellow表达。在过氧化氢预处理后短暂产生的活性氧（ROS）的作用下，cerevisia在线粒体部位发出明显的黄色荧光。通过y1 -黄色荧光观察，我们还发现，在氰化物的呼吸抑制作用下，线粒体在染色体DNA周围重排形成簇状结构，随后产生ROS。相反，氧化磷酸化解偶联剂的摄取与线粒体重排无关。这些结果表明，y1 -黄色在活细胞中线粒体活力和形态可视化中的实用性。
We have visualized redox and structural changes in the mitochondria of yeastSaccharomyces cerevisiaeas a eukaryotic cell model using a genetically encoded yellow fluorescent protein (Y1-Yellow) and conventional fluorescence microscopy. Y1-Yellow originating from a yellow emitting luminous bacteriumAliivibrio sifiaeY1 was fused with a mitochondria-targeted sequence (mt-sequence). Y1-Yellow fluorescence arising only from the mitochondrial site and the color of yellow fluorescence could be easily differentiated from cellular autofluorescence and from that of conventional probes. Y1-Yellow expressingS. cerevisiaemade the yellow fluorescence conspicuous at the mitochondrial site in response to reactive oxygen species (ROS) transiently derived in the wake of pretreatment with hydrogen peroxide. Based on our observation with Y1-Yellow fluorescence, we also showed that mitochondria rearrange to form a cluster structure surrounding chromosomal DNA via respiratory inhibition by cyanide, followed by the generation of ROS. In contrast, uptake of an uncoupler of oxidative phosphorylation is not responsible for mitochondrial rearrangement. These results indicate the utility of Y1-Yellow for visualization of mitochondrial vitality and morphology in living cells.