title: Actomyosin II contractility expels von Willebrand factor from Weibel-Palade bodies during exocytosis.
abstract: The study of actin in regulated exocytosis has a long history with many different results in numerous systems. A major limitation on identifying precise mechanisms has been the paucity of experimental systems in which actin function has been directly assessed alongside granule content release at distinct steps of exocytosis of a single secretory organelle with sufficient spatiotemporal resolution. Using dual-color confocal microscopy and correlative electron microscopy in human endothelial cells, we visually distinguished two sequential steps of secretagogue-stimulated exocytosis: fusion of individual secretory granules (Weibel-Palade bodies [WPBs]) and subsequent expulsion of von Willebrand factor (VWF) content. Based on our observations, we conclude that for fusion, WPBs are released from cellular sites of actin anchorage. However, once fused, a dynamic ring of actin filaments and myosin II forms around the granule, and actomyosin II contractility squeezes VWF content out into the extracellular environment. This study therefore demonstrates how discrete actin cytoskeleton functions within a single cellular system explain actin filament-based prevention and promotion of specific exocytic steps during regulated secretion.
authors.affiliation: Medical Research Council Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, England, UK.
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Nightingale
authors.forename: Ian J
authors.initials: IJ
authors.lastname: White
authors.forename: Emily L
authors.initials: EL
authors.lastname: Doyle
authors.forename: Mark
authors.initials: M
authors.lastname: Turmaine
authors.forename: Kimberly J
authors.initials: KJ
authors.lastname: Harrison-Lavoie
authors.forename: Kathleen F
authors.initials: KF
authors.lastname: Webb
authors.forename: Louise P
authors.initials: LP
authors.lastname: Cramer
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Cutler
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000205:Actomyosin / Q000378:metabolism*; D002478:Cells, Cultured; D003572:Cytochalasins / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D005089:Exocytosis / Q000187:drug effects; D006801:Humans; D008561:Membrane Fusion; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D018715:Microscopy, Video; D024462:Myosin Type II / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D020302:Weibel-Palade Bodies / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D014841:von Willebrand Factor / Q000378:metabolism*
chemical_list: D003572:Cytochalasins; D011993:Recombinant Fusion Proteins; D014841:von Willebrand Factor; C007215:cytochalasin E; D000205:Actomyosin; D024462:Myosin Type II