Super-resolution imaging of the dynamic cleavage of intercellular tunneling nanotubes
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RESEARCH ARTICLE
Super-resolution imaging of the dynamic cleavage of intercellular tunneling nanotubes Wanjun GONG#, Wenhui PAN#, Ying HE, Meina HUANG, Jianguo ZHANG, Zhenyu GU, Dan ZHANG, Zhigang YANG (✉), Junle QU
Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China
© Higher Education Press 2020
Abstract As a new method of cell–cell communication, tunneling nanotubes (TNTs) play important roles in cell– cell signaling and mass exchanges. However, a lack of powerful tools to visualize dynamic TNTs with high temporal/spatial resolution restricts the exploration of their formation and cleavage, hindering the complete understanding of its mechanism. Herein, we present the first example of using stochastic optical reconstruction microscopy (STORM) to observe the tube-like structures of TNTs linking live cells with an easily prepared fluorescent dye. Because of this new imaging microscopy, the cleavage process of TNTs was observed with a high spatial resolution. Keywords super-resolution, tunneling nanotubes (TNTs), live cell
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Introduction
Intercellular communication is essential for tissue homeostasis, specific cell functions, and responses to external stimulus. Cell–cell communications typically occur through various processes, such as diffusible factors, including cytokines and chemokines, secreted microvesicles, and direct passage through gap junctions [1]. Recently, a new impressive intercellular communication pathway was found that allows long-distance cell–cell contacts via tunneling nanotubes (TNTs) between these cells, which was initially reported in rat pheochromocytoma (PC12)-derived cells [2] and later in other types of cells [2–7]. The TNTs connecting two cells exhibit long Received July 18, 2020; accepted September 11, 2020 E-mail: [email protected] #
These authors contributed equally to this work.
tubular structures with diameters of 50–1500 nm and lengths of tens to hundreds of microns, which maintain a continuous cytoplasm membrane between the connecting cells, thus allowing the transportation of numerous cellular components, such as proteins, RNAs, viruses, and organelles, from one cell to another [8–10]. Generally, actin is included in the central composition and a membrane is wrapped outside, which is typically part of the cell membrane. Additionally, TNTs play distinct roles in the modulation of cell death involved not only in the delivery of injured cells [9–12] but also in enhancing the lysis of distant cells [13]. Thus, addressing the dynamic changes of such TNTs will be helpful for understanding the exact events of intercellular communication behaviors. In recent years, studies on TNTs were performed in normal cells as well as on the pathogenesis of neurodegenerative diseases and cancer [14] mainly via electron and fluorescence microscopy [11,15–19]. However, it is difficult to observe ultra-fine TNTs in vivo because of their fragility to light, me
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