期刊
JOURNAL OF THROMBOSIS AND HAEMOSTASIS
卷 12, 期 4, 页码 519-527出版社
WILEY
DOI: 10.1111/jth.12513
关键词
blood platelets; laminin; platelet activation; cell-derived microparticles; exosomes
资金
- Swedish Cancer Society
- Karolinska Institutet [ETF8932]
- Estonian Science Agency
- EMBO Installation Grant
Background Blood platelets secrete upon activation of laminins 411/421 and 511/521, large adhesive proteins mainly found in the basement membranes of blood vessels and other tissues. At present, the subcellular localization and secretion mechanisms of platelet laminins are largely unknown. Objectives Our aim was to compare the subcellular localization of laminins 411/421 and 511/521 and specific granule markers in platelets. We also elucidated the role of microvesicles and exosomes in laminin release in platelet activation. Methods We studied laminin and granule marker protein localization in platelets by using immunofluorescence confocal microscopy and immunoelectron microscopy. Microvesicles and exosomes were separated from material released from platelets on activation by thrombin. The expression of laminins in microvesicles and exosomes was studied by using SDS-PAGE and Western blotting as well as by flow cytometric analysis. The exosomes were immunoprecipitated with magnetic microbeads coated with anti-CD63 antibodies. Results and conclusions We demonstrate that laminins 411/421 and 511/521 are present in compartments of platelets that do not express alpha-granule, dense granule, or lysosome marker proteins. Moreover, laminins secreted by activated platelets are mostly found in microvesicles shed from the plasma membrane, while their presence in simultaneously released exosomes is minimum.
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