Oikawa Kazusato, Imai Takuto, Kodama Yutaka, Numata Keiji
Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Kyoto-Daigaku-Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Biomacromolecules Research Team, RIKEN Center for Sustainable Resource Science, Wako, Saitama 351-0198, Japan.
Plant Biotechnol (Tokyo). 2021 Jun 25;38(2):257-262. doi: 10.5511/plantbiotechnology.21.0204a.
Mitochondria-selective fluorescent probes such as MitoTracker are often used for mitochondria imaging in various plants. Although some of the probes are reported to induce mitochondria dysfunction in animal cells, the effect on plant cells remains to be determined. In the present study, we applied quantitative methods to analyze mitochondrial movement, speed frequency, and speed-angle changes, based on trajectory analysis of mitochondria in mesophyll protoplast cells of expressing the mitochondria-localized fluorescent protein. Using the quantitative method, we assessed whether MitoTracker Red (FM and CMXRos) induce mitochondria dysfunction in . Although both the fluorescent probes well-stained mitochondria, the CMXRos probe, not the FM probe, gave a severe effect on mitochondrial movement at the low concentration (10 nM), indicating a MitoTracker-induced mitochondria dysfunction in . These results revealed that our quantitative method based on mitochondrial movement can be used to determine the appropriate concentrations of mitochondria-selective fluorescent probes in plants.
线粒体选择性荧光探针,如MitoTracker,常用于各种植物的线粒体成像。尽管有报道称某些探针会在动物细胞中诱导线粒体功能障碍,但对植物细胞的影响仍有待确定。在本研究中,我们基于表达线粒体定位荧光蛋白的叶肉原生质体细胞中线粒体的轨迹分析,应用定量方法分析线粒体的运动、速度频率和速度角度变化。使用该定量方法,我们评估了MitoTracker Red(FM和CMXRos)是否会在[具体植物或细胞类型未明确]中诱导线粒体功能障碍。尽管两种荧光探针都能很好地对线粒体进行染色,但CMXRos探针而非FM探针在低浓度(10 nM)时对线粒体运动产生了严重影响,表明MitoTracker在[具体植物或细胞类型未明确]中诱导线粒体功能障碍。这些结果表明,我们基于线粒体运动的定量方法可用于确定植物中线粒体选择性荧光探针的合适浓度。