Dynamic single-vesicle tracking of cell-bound membrane vesicles on resting, activated, and cytoskeleton-disrupted cells
Dynamic single-vesicle tracking of cell-bound membrane vesicles on resting, activated, and cytoskeleton-disrupted cells
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对静息细胞、激活细胞和细胞骨架破坏细胞上的细胞结合膜囊泡进行动态单囊泡追踪
DOI：
10.1016/j.bbamem.2018.10.013
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发表时间：
2019
影响因子：
3.4
通讯作者：
Chen Yong
中科院分区：
文献类型：
--
作者：
Zhang Wendiao;Xu Ye;Chen Guo;Wang Kun;Shan Wenzhe;Chen Yong
细胞质膜中预先存在的细胞结合膜囊泡的组成、结构、产生、运动、命运和功能知之甚少。在这里，单个囊泡跟踪的细胞结合的膜囊泡在用或不用各种试剂处理的内皮细胞的质膜进行调查的细胞结合的膜囊泡的运动。在单囊泡追踪之前确认了这些试剂中每种试剂的功效。通过单囊泡追踪，我们发现oxLDL、TNF-α和VEGFα显著增加了每个细胞的细胞结合膜囊泡的平均数量，这意味着oxLDL、TNF-α和VEGFα激活细胞可以触发细胞结合膜囊泡的产生。oxLDL、TNF-α、VEGFα、LPS和MβCD均能显著影响细胞结合膜的运动速度，但对细胞结合膜囊泡的位移（运动范围）无显著影响。单囊泡跟踪进一步揭示，每个细胞的细胞结合的膜囊泡的平均数量和单个细胞结合的膜囊泡的平均速度/位移可以显着改变的细胞分裂素破坏试剂（细胞松弛素D和诺考达唑）。这些数据表明，细胞结合膜囊泡的产生和运动可能是由细胞内的细胞骨架控制，并能够受到多种条件的影响，如细胞活化，膜流动性改变，和其他。
The composition, structure, production, motion, fate, and functions of cell-bound membrane vesicles pre-existing in the plasma membrane of cells are poorly understood. Here, single-vesicle tracking of individual cell-bound membrane vesicles in the plasma membrane of endothelial cells treated with or without various reagents was performed to investigate the motion of cell-bound membrane vesicles. The efficacy of each of these reagents was confirmed prior to single-vesicle tracking. Via single-vesicle tracking, we found that oxLDL, TNF-α, and VEGFα significantly increased the average number of cell-bound membrane vesicles per cell, implying that cell activation by oxLDL, TNF-α, and VEGFα could trigger the production of cell-bound membrane vesicles. It was also found that oxLDL, TNF-α, VEGFα, LPS, and MβCD but not LDL could significantly affect the motion speed of cell-bound membrane whereas none of them could significantly influence the displacement (moving range) of cell-bound membrane vesicles. The single-vesicle tracking further revealed that the average number of cell-bound membrane vesicles per cell and the mean speed/displacement of individual cell-bound membrane vesicles could be dramatically altered by the cytoskeleton-disrupting reagents (cytochalasin D and nocodazole). The data imply that the production and movement of cell-bound membrane vesicles are probably controlled by intracellular cytoskeletons and capable of being affected by multiple conditions e.g. cell activation, membrane fluidity alteration, and others.