Triggered Cell-Cell Fusion Assay forCytoplasmic and Organelle IntermixingStudies

Daniel Feliciano; Jonathon Nixon-Abell; Jennifer Lippincott-Schwartz

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Triggered Cell-Cell Fusion Assay forCytoplasmic and Organelle IntermixingStudies

机译：触发的细胞 - 细胞融合测定为单独的和细胞器膜混合术

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摘要

Different multicellular organisms undergo cell-cell fusion to form functionalsyncytia that support specialized functions necessary for proper developmentand survival. For years, monitoring the structural consequences of this processusing live-cell imaging has been challenging due to the unpredictable timingof cell fusion events in tissue systems. Here we present a triggered vesicularstomatitis virus G-protein (VSV-G)-mediated cell-cell fusion assay that canbe used to synchronize fusion between cells. This allows the study of cellularchanges that occur during cell fusion. The process is induced using a fast washof low pH isotonic buffer, promoting the fusion of plasma membranes of twoor more adjacent cells within seconds. This approach is suitable for studyingmixing of small cytoplasmic molecules between fusing cells as well as changesin organelle distribution and dynamics.

机译：不同的多细胞生物经历细胞 - 细胞融合以形成支持适当发育和存活所必需的专业功能的官能。多年来，由于组织系统中的不可预测的细胞融合事件，监测该处理活细胞成像的结构后果一直在挑战。在这里，我们提出了一种触发的血管瘤性炎病毒G蛋白（VSV-G）介导的细胞 - 细胞融合测定，其可以用于同步细胞之间的融合。这允许研究在细胞融合期间发生的细胞硬化。使用低pH异位缓冲液的快洗诱导该方法，在几秒钟内促进多个相邻细胞的血浆膜的熔化。这种方法适用于融合细胞之间的小型细胞质分子以及变化细胞器分布和动力学的研究。

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来源
《Current Protocols in Cell Biology》 |2018年第1期|共13页
作者
Daniel Feliciano; Jonathon Nixon-Abell; Jennifer Lippincott-Schwartz;
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作者单位

Janelia Research Campus Howard Hughes Medical Institute Ashburn Virginia;

Janelia Research Campus Howard Hughes Medical Institute Ashburn Virginia;

Janelia Research Campus Howard Hughes Medical Institute Ashburn Virginia;

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原文格式 PDF
正文语种 eng
中图分类细胞生物学;
关键词
cell-cell fusion; stable cell lines; synchronized fusion; syncytia; vesicular stomatitis virus G-protein (VSV-G);

机译：细胞 - 细胞融合;稳定的细胞系;同步融合;Syncytia;囊泡口腔炎病毒G-蛋白（VSV-G）;

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专利

1. 胸腺细胞和细胞器水平的线粒体膜电势研究 [J] . 王通, 曾耀英, 邢飞跃, 分子细胞生物学报（英文版） . 2006,第002期
2. Triggered Cell-Cell Fusion Assay forCytoplasmic and Organelle IntermixingStudies [J] . Daniel Feliciano, Jonathon Nixon-Abell, Jennifer Lippincott-Schwartz Current Protocols in Cell Biology . 2018,第1期

机译：触发的细胞 - 细胞融合测定为单独的和细胞器膜混合术
3. Identification of Nafamostat as a Potent Inhibitor of Middle East Respiratory Syndrome Coronavirus S Protein-Mediated Membrane Fusion Using the Split-Protein-Based Cell-Cell Fusion Assay [J] . Yamamoto Mizuki, Matsuyama Shutoku, Li Xiao, Antimicrobial agents and chemotherapy. . 2016,第11期

机译：使用基于分裂蛋白的细胞-细胞融合测定法鉴定Nafamostat作为中东呼吸系统综合症冠状病毒S蛋白介导的膜融合的有效抑制剂。
4. Ability of antibodies specific to the HIV-1 envelope glycoprotein to block the fusion inhibitor T20 in a cell-cell fusion assay [J] . VincentN., MalvoisinE. Immunobiology: Zeitschrift fur Immunitatsforschung . 2012,第10期

机译：HIV-1包膜糖蛋白特异性抗体在细胞-细胞融合测定中阻断融合抑制剂T20的能力
5. Spatial and temporal assays to determine the dynamics of protein localisation and organelle movement in single living cells [C] . R.J. Errington, P.J. Smith, S.C.Chappell, Microscopy Society of America annual meeting;Microscopy Society of Canada annual meeting;Societe de Microscopie du Canada annual meeting;Microbeam Analysis Society annual meeting;International Metallographic Society annual meeting . 2002

机译：确定单个活细胞中蛋白质定位和细胞器运动动态的时空分析
6. Pushing the Envelope: How HIV Regulates Dual Roles for Viral ENV Glycoproteins in Cell-Cell Adhesion and Membrane Fusion [D] . Gardiner, Jaye Cassandra 2017

机译：推信封：HIV如何调节病毒ENV糖蛋白在细胞-细胞粘附和膜融合中的双重作用
7. Identification of Nafamostat as a Potent Inhibitor of Middle East Respiratory Syndrome Coronavirus S Protein-Mediated Membrane Fusion Using the Split-Protein-Based Cell-Cell Fusion Assay [O] . Mizuki Yamamoto, Shutoku Matsuyama, Xiao Li, 2016

机译：使用基于分裂蛋白的细胞-细胞融合测定法鉴定Nafamostat作为中东呼吸系统综合症冠状病毒S蛋白介导的膜融合的有效抑制剂。
8. Lipid Mixing Assay for Murine Myoblast Fusion and Other Slow Cell-cell Fusion Processes [O] . Evgenia Leikina, Kamran Melikov, Anthony Rabinovich, 2020

机译：用于鼠肌细胞融合和其他缓慢细胞 - 细胞融合方法的脂质混合测定

Triggered Cell-Cell Fusion Assay forCytoplasmic and Organelle IntermixingStudies

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