Han Junyan, Han Myung Shin, Tung Ching-Hsuan
Department of Translational Imaging, The Methodist Hospital Research Institute, Weill Cornell Medical College, Houston, TX, USA.
Biochim Biophys Acta. 2013 Nov;1830(11):5130-5. doi: 10.1016/j.bbagen.2013.07.001. Epub 2013 Jul 12.
Mitochondria, powerhouses of cells, are responsible for many critical cellular functions, such as cell energy metabolism, reactive oxygen species production, and apoptosis regulation. Monitoring mitochondria morphology in live cells temporally and spatially could help with the understanding of the mechanisms of mitochondrial functional regulation and the pathogenesis of mitochondria-related diseases.
A novel non-cytotoxic fluorogenic compound, AcQCy7, was developed as a mitochondria-specific dye.
AcQCy7 emitted no fluorescent signal outside of cells, but it became fluorescent after intracellular hydrolysis of the acetyl group. The hydrolyzed fluorescent product was well retained in mitochondria, enabling long-lasting fluorescence imaging of mitochondria without cell washing. A 2-day culture study using AcQCy7 showed no sign of cytotoxicity, whereas a commonly used mitochondria-staining probe, Mitochondria Tracker Green, caused significant cell death even at a much lower concentration. Apoptosis-causing mitochondria fission was monitored clearly in real time by AcQCy7.
A simple add-and-read mitochondria specific dye AcQCy7 has been validated in various cell models. Bright mitochondria specific fluorescent signal in treated cells lasted several days without noticeable toxicity.
The probe AcQCy7 has been proofed to be a non-toxic agent for long-term mitochondria imaging.
线粒体作为细胞的“动力工厂”,负责许多关键的细胞功能,如细胞能量代谢、活性氧生成及凋亡调控。在活细胞中对线粒体形态进行时空监测,有助于理解线粒体功能调控机制及线粒体相关疾病的发病机制。
开发了一种新型无细胞毒性的荧光化合物AcQCy7作为线粒体特异性染料。
AcQCy7在细胞外不发出荧光信号,但在细胞内乙酰基水解后会发出荧光。水解后的荧光产物能很好地保留在线粒体中,无需洗细胞即可对线粒体进行长时间的荧光成像。使用AcQCy7进行的为期2天的培养研究未显示出细胞毒性迹象,而常用的线粒体染色探针Mitochondria Tracker Green即使在低得多的浓度下也会导致显著细胞死亡。AcQCy7能实时清晰监测导致凋亡的线粒体分裂。
一种简单的即加即读型线粒体特异性染料AcQCy7已在多种细胞模型中得到验证。处理后的细胞中明亮的线粒体特异性荧光信号可持续数天且无明显毒性。
探针AcQCy7已被证明是一种用于长期线粒体成像的无毒试剂。