Feliciano Daniel, Nixon-Abell Jonathon, Lippincott-Schwartz Jennifer
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia.
Curr Protoc Cell Biol. 2018 Dec;81(1):e61. doi: 10.1002/cpcb.61. Epub 2018 Aug 13.
Different multicellular organisms undergo cell-cell fusion to form functional syncytia that support specialized functions necessary for proper development and survival. For years, monitoring the structural consequences of this process using live-cell imaging has been challenging due to the unpredictable timing of cell fusion events in tissue systems. Here we present a triggered vesicular stomatitis virus G-protein (VSV-G)-mediated cell-cell fusion assay that can be used to synchronize fusion between cells. This allows the study of cellular changes that occur during cell fusion. The process is induced using a fast wash of low pH isotonic buffer, promoting the fusion of plasma membranes of two or more adjacent cells within seconds. This approach is suitable for studying mixing of small cytoplasmic molecules between fusing cells as well as changes in organelle distribution and dynamics. © 2018 by John Wiley & Sons, Inc.
不同的多细胞生物会经历细胞间融合以形成功能性合胞体，这些合胞体支持正常发育和生存所需的特殊功能。多年来，由于组织系统中细胞融合事件的时间不可预测，利用活细胞成像监测这一过程的结构后果一直具有挑战性。在此，我们展示了一种触发型水泡性口炎病毒G蛋白（VSV-G）介导的细胞间融合测定法，该方法可用于同步细胞间的融合。这使得研究细胞融合过程中发生的细胞变化成为可能。该过程通过快速冲洗低pH等渗缓冲液来诱导，可在数秒内促进两个或更多相邻细胞质膜的融合。这种方法适用于研究融合细胞间小细胞质分子的混合以及细胞器分布和动态变化。© 2018约翰威立国际出版公司
