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• W2783004471 abstract "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 analysis demonstrates that MVB-PM fusion frequency is reduced by depleting the target membrane SNAREs SNAP23 and syntaxin-4 but also can be induced in single cells by stimulation of the histamine H1 receptor (H1HR). Interestingly, activation of H1R1 in HeLa cells increases Ser110 phosphorylation of SNAP23, promoting MVB-PM fusion and the release of CD63-enriched exosomes. Using this single-cell resolution approach, we highlight the modulatory dynamics of MVB exocytosis that will help to increase our understanding of exosome physiology and identify druggable targets in exosome-associated pathologies." @default.
• W2783004471 created "2018-01-26" @default.
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• W2783004471 date "2018-01-16" @default.
• W2783004471 modified "2023-10-13" @default.
• W2783004471 title "Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling" @default.
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• W2783004471 doi "https://doi.org/10.1083/jcb.201703206" @default.
• W2783004471 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5839777" @default.
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• W2783004471 hasPublicationYear "2018" @default.
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