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

Autor: Juan J. Garcia-Vallejo, D. Michiel Pegtel, Connie R. Jimenez, Martine J. Smit, Anoek Zomer, Frederik J. Verweij, Marc G. Coppolino, Graça Raposo, S. Rubina Baglio, Hans Janssen, Jaco C. Knol, Jacques Neefjes, Sander R. Piersma, Jacco van Rheenen, Maarten P. Bebelman, Matthijs Verhage, Guillaume van Niel, Jaap M. Middeldorp, Richard de Goeij-de Haas, Ruud F. Toonen, Ilse Hurbain
Přispěvatelé: van Niel, Guillaume, VU University Medical Center [Amsterdam], Biologie Cellulaire et Cancer, Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), BioImaging Cell and Tissue Core Facility (PICT-IBiSA), Institut Curie [Paris], Netherlands Cancer Institute (NKI), Antoni van Leeuwenhoek Hospital, Leiden University Medical Center (LUMC), Hubrecht Institute [Utrecht, Netherlands], University Medical Center [Utrecht]-Royal Netherlands Academy of Arts and Sciences (KNAW), University Medical Center [Utrecht], University of Guelph, Vrije Universiteit Amsterdam [Amsterdam] (VU), Functional Genomics, Amsterdam Neuroscience - Cellular & Molecular Mechanisms, Medicinal chemistry, AIMMS, Pathology, CCA - Imaging and biomarkers, Medical oncology laboratory, AGEM - Re-generation and cancer of the digestive system, Molecular cell biology and Immunology, Human genetics, Vrije universiteit = Free university of Amsterdam [Amsterdam] (VU), VU University Amsterdam
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Zdroj: Verweij, F J, Bebelman, M P, Jimenez, C R, Garcia-Vallejo, J J, Janssen, H, Neefjes, J, Knol, J C, de Goeij-de Haas, R, Piersma, S R, Baglio, S R, Verhage, M, Middeldorp, J M, Zomer, A, van Rheenen, J, Coppolino, M G, Hurbain, I, Raposo, G, Smit, M J, Toonen, R F G, van Niel, G & Pegtel, D M 2018, ' Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling ', Journal of Cell Biology, vol. 217, no. 3, pp. 1129-1142 . https://doi.org/10.1083/jcb.201703206
The Journal of Cell Biology
Journal of Cell Biology
Journal of Cell Biology, 2018, 217 (3), pp.1129-1142. ⟨10.1083/jcb.201703206⟩
Journal of Cell Biology, 217(3), 1129-1142. Rockefeller University Press
Journal of Cell Biology, 217(3), 1129. Rockefeller University Press
Journal of Cell Biology, Rockefeller University Press, 2018, 217 (3), pp.1129-1142. ⟨10.1083/jcb.201703206⟩
Journal of Cell Biology, 217(3), 1129-1142
ISSN: 0021-9525
1540-8140
DOI: 10.1083/jcb.201703206
Popis: All mammalian cells release small endosome-derived exosomes that function in intercellular communication, but the secretion process is poorly understood. Verweij et al. developed a live-imaging approach and demonstrate that external cues can trigger exosome release from a subpopulation of multivesicular bodies by phosphorylating the target membrane SNARE SNAP23 at serine residue 110.
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.
Databáze: OpenAIRE
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