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Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling.

J. Cell Biol.. 2018; 
Verweij Frederik Johannes,Bebelman Maarten P,Jimenez Connie R,Garcia-Vallejo Juan J,Janssen Hans,Neefjes Jacques,Knol Jaco C,de Goeij-de Haas Richard,Piersma Sander R,Baglio S Rubina,Verhage Matthijs,Middeldorp Jaap M,Zomer Anoek,van Rheenen Jacco,Coppolino Marc G,Hurbain Ilse,Raposo Gra?a,Smit Martine J,Toonen Ruud F G,van Niel Guillaume,Pegtel D Mic
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摘要

Exosomes are small endosome-derived extracellular vesicles implicated in cell-cell communication and are secreted by living cells when multivesicular bodies (MVBs) fuse with the plasma membrane (PM). Current techniques to study exosome physiology are based on isolation procedures after secretion, precluding direct and dynamic insight into the mechanics of exosome biogenesis and the regulation of their release. In this study, we propose real-time visualization of MVB-PM fusion to overcome these limitations. We designed tetraspanin-based pH-sensitive optical reporters that detect MVB-PM fusion using live total internal reflection fluorescence and dynamic correlative light-electron microscopy. Quantitative ana... More

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