The role of platelet and endothelial GARP in thrombosis and hemostasis.
The role of platelet and endothelial GARP in thrombosis and hemostasis.
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DOI：
10.1371/journal.pone.0173329
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Tersteeg C
中科院分区：
文献类型：
--
作者：
Vermeersch E;Denorme F;Maes W;De Meyer SF;Vanhoorelbeke K;Edwards J;Shevach EM;Unutmaz D;Fujii H;Deckmyn H;Tersteeg C
糖蛋白-A重复主要蛋白（GARP或LRRC 32）存在于人血小板和内皮细胞等上。斑马鱼中GARP的完全敲除表明其参与血栓形成的证据。使用血小板和内皮条件性GARP敲除小鼠评价GARP在血小板生理学和血栓形成中的作用。使用Cre-loxP重组系统产生血小板和内皮特异性GARP敲除小鼠。用流式细胞术、铺展分析法和PAR 4活化肽及胶原相关肽聚集测定法检测去GARP血小板的功能。此外，分析了流动下的凝块收缩和胶原诱导的血小板粘附和聚集。最后，在体内尾部出血时间，闭塞时间的肠系膜和颈动脉后FeCl 3诱导血栓形成的血小板和内皮细胞特异性GARP基因敲除小鼠进行了测定。血小板特异性GARP基因敲除小鼠表面GPIb、GPVI和整合素αIIb糖蛋白表达正常。尽管血小板活化后GARP表达增加，但无GARP的血小板显示正常激动剂诱导的活化、在纤维蛋白原上扩散和聚集反应。此外，血小板上GARP的缺失不影响凝块收缩，并且对流动下胶原涂覆表面上的血栓形成没有影响。与此一致，在血小板或内皮特异性GARP敲除小鼠中，FeCl 3诱导血栓形成后，颈动脉和肠系膜动脉中的尾部出血时间和闭塞时间均未受到影响。提供了血小板和内皮GARP在小鼠中的止血和血栓形成中不重要的证据。
Glycoprotein-A Repetitions Predominant protein (GARP or LRRC32) is present on among others human platelets and endothelial cells. Evidence for its involvement in thrombus formation was suggested by full knockout of GARP in zebrafish. To evaluate the role of GARP in platelet physiology and in thrombus formation using platelet and endothelial conditional GARP knock out mice. Platelet and endothelial specific GARP knockout mice were generated using the Cre-loxP recombination system. The function of platelets without GARP was measured by flow cytometry, spreading analysis and aggregometry using PAR4-activating peptide and collagen related peptide. Additionally, clot retraction and collagen-induced platelet adhesion and aggregation under flow were analyzed. Finally, in vivo tail bleeding time, occlusion time of the mesenteric and carotid artery after FeCl3-induced thrombosis were determined in platelet and endothelial specific GARP knock out mice. Platelet specific GARP knockout mice had normal surface GPIb, GPVI and integrin αIIb glycoprotein expression. Although GARP expression was increased upon platelet activation, platelets without GARP displayed normal agonist induced activation, spreading on fibrinogen and aggregation responses. Furthermore, absence of GARP on platelets did not influence clot retraction and had no impact on thrombus formation on collagen-coated surfaces under flow. In line with this, neither the tail bleeding time nor the occlusion time in the carotid- and mesenteric artery after FeCl3-induced thrombus formation in platelet or endothelial specific GARP knock out mice were affected. Evidence is provided that platelet and endothelial GARP are not important in hemostasis and thrombosis in mice.