dc.contributor.author | NIGHTINGALE, TD | |
dc.contributor.author | McCormack, JJ | |
dc.contributor.author | Grimes, W | |
dc.contributor.author | ROBINSON, C | |
dc.contributor.author | Lopes da Silva, M | |
dc.contributor.author | White, IJ | |
dc.contributor.author | Vaughan, A | |
dc.contributor.author | Cramer, LP | |
dc.contributor.author | Cutler, DF | |
dc.date.accessioned | 2018-07-26T09:10:57Z | |
dc.date.available | 2018-07-26T09:10:57Z | |
dc.date.issued | 2018-06-28 | |
dc.date.submitted | 2018-07-09T16:11:00.429Z | |
dc.identifier.issn | 1538-7836 | |
dc.identifier.uri | http://qmro.qmul.ac.uk/xmlui/handle/123456789/42604 | |
dc.description.abstract | BACKGROUND:
Endothelial cells harbour specialised storage organelles, Weibel-Palade Bodies (WPBs). Exocytosis of WPB content into the vascular lumen initiates primary haemostasis, mediated by Von Willebrands factor (VWF) and inflammation, mediated by several proteins including P-selectin. During full fusion, secretion of this large haemostatic protein and smaller pro-inflammatory proteins are thought to be inextricably linked.
OBJECTIVE:
To determine if secretagogue-dependent differential release of WPB cargo occurs, and whether this is mediated by the formation of an actomyosin ring during exocytosis.
METHODS:
We used VWF string analysis, leukocyte rolling assays, ELISA, spinning disk confocal microscopy, high-throughput confocal microscopy and inhibitor and siRNA treatments to demonstrate the existence of cellular machinery that allows differential release of WPB cargo proteins.
RESULTS:
Inhibition of the actomyosin ring differentially effects two processes regulated by WPB exocytosis; it perturbs VWF string formation but has no effect on leukocyte rolling. The efficiency of ring recruitment correlates with VWF release; the ratio of release of VWF to small cargoes decreases when ring recruitment is inhibited. The recruitment of the actin ring is time-dependent; fusion events occurring directly after stimulation are less likely to initiate haemostasis than later events, and is activated by PKC isoforms.
CONCLUSIONS:
Secretagogues differentially recruit the actomyosin ring, thus demonstrating one mechanism by which the pro-thrombotic effect of endothelial activation can be modulated. This potentially limits thrombosis whilst permitting a normal inflammatory response. These results have implications for the assessment of WPB fusion, cargo-content release and the treatment of patients with von Willebrand disease. | en_US |
dc.description.sponsorship | T. D. Nightingale was funded by a Medical Research Councilproject grant MR/M019179/1and a British Heart Foundationproject grant (PG/15/72/31732). J.J. McCormack and D.F. Cutler were funded by an MRC programme grant MC_UU_12018/2. I.J. White was funded by MRC LMCB core. W. Grimes was supported partlyby the Biomedical Research Council of A*STAR (Agency for Science, Technology and Research), Singapore, partly by the MRC LMCB. C. Robinson was funded by a BHF project grant (PG/15/72/31732).M. Lopes da Silva reports grants from MRC during the conduct of the study; and grants from MRC outside the submitted work. The other authors state that they have no conflict of interest. | en_US |
dc.language.iso | en | en_US |
dc.publisher | Wiley | en_US |
dc.relation.ispartof | Journal of Thrombosis and Haemostasis | |
dc.relation.isreplacedby | 123456789/45643 | |
dc.relation.isreplacedby | http://qmro.qmul.ac.uk/xmlui/handle/123456789/45643 | |
dc.rights | This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: 10.1111/jth.14218. This article is protected by copyright. All rights reserved. | |
dc.title | Tuning the endothelial response: differential release of exocytic cargos from Weibel-Palade Bodies. | en_US |
dc.type | Article | en_US |
dc.identifier.doi | 10.1111/jth.14218. | |
pubs.publication-status | Published online | |
pubs.publisher-url | https://doi.org/10.1111/jth.14218 | |