The State Key Lab of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Chemistry and Biomedicine Innovation Center (ChemBIC), Nanjing University, Nanjing, Jiangsu, 210093, China.
School of Chemical Sciences, University of Chinese Academy of Science, Beijing, 100190, China.
Angew Chem Int Ed Engl. 2023 Aug 21;62(34):e202303053. doi: 10.1002/anie.202303053. Epub 2023 Jul 12.
The in-depth study of single cells requires the dynamically molecular information in one particular nanometer-sized organelle in a living cell, which is difficult to achieve using current methods. Due to high efficiency of click chemistry, a new nanoelectrode-based pipette architecture with dibenzocyclooctyne at the tip is designed to realize fast conjugation with azide group-containing triphenylphosphine, which targets mitochondrial membranes. The covalent binding of one mitochondrion at the tip of the nanopipette allows a small region of the membrane to be isolated on the Pt surface inside the nanopipette. Therefore, the release of reactive oxygen species (ROS) from the mitochondrion is monitored, which is not interfered by the species present in the cytosol. The dynamic tracking of ROS release from one mitochondrion reveals the distinctive "ROS-induced ROS release" within the mitochondria. Further study of RSL3-induced ferroptosis using nanopipettes provides direct evidence for supporting the noninvolvement of glutathione peroxidase 4 in the mitochondria during RSL3-induced ROS generation, which has not previously been observed at the single-mitochondrion level. Eventually, this established strategy should overcome the existing challenge of the dynamic measurement of one special organelle in the complicated intracellular environment, which opens a new direction for electroanalysis in subcellular analysis.
深入研究单细胞需要在活细胞中动态获取特定纳米大小细胞器内的分子信息，而这一点目前很难实现。由于点击化学的高效性，设计了一种新型基于纳米电极的尖端带有二苯并环辛炔的玻璃管电极结构，用于实现与含有叠氮基团的三苯基膦的快速结合，该三苯基膦靶向线粒体膜。纳米管尖端处的一个线粒体的共价结合允许将膜的一小部分隔离在纳米管内的 Pt 表面上。因此，可以监测来自线粒体的活性氧物质（ROS）的释放，这不会受到细胞质中存在的物质的干扰。从一个线粒体中释放 ROS 的动态追踪揭示了线粒体内部独特的“ROS 诱导的 ROS 释放”。使用纳米管对 RSL3 诱导的铁死亡进行进一步研究，为支持谷胱甘肽过氧化物酶 4 不参与 RSL3 诱导的 ROS 生成提供了直接证据，这在以前的单线粒体水平上尚未观察到。最终，这种已建立的策略应该能够克服在复杂的细胞内环境中对一个特殊细胞器进行动态测量的现有挑战，为亚细胞分析中的电分析开辟了新的方向。
