Real-time analysis of platelet aggregation and procoagulant activity during thrombus formation in vivo
Real-time analysis of platelet aggregation and procoagulant activity during thrombus formation in vivo
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DOI：
10.1007/s00424-008-0466-9
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发表时间：
2008-09-01
影响因子：
4.5
通讯作者：
Urano, Tetsumei
中科院分区：
文献类型：
--
作者：
Hayashi, Tadataka;Mogami, Hideo;Urano, Tetsumei
血管血栓形成的确切机制仍有待确定。在这里，我们介绍了一种新的方法来探测血栓形成的活体动物血管中使用活体显微镜在绿色荧光蛋白（GFP）转基因小鼠，同时监测血小板聚集和促凝血活性。为此，使用荧光染料标记的表达GFP的血小板和膜联蛋白A5来可视化和分析血小板聚集和促凝血活性的标志物（血小板表面磷脂酰丝氨酸（PS））。在活体动物血管中，激光诱导的血栓随着时间的推移而增大，然后减小，而血小板表面PS起始于损伤部位，然后渗透到血栓中。PS阳性血小板主要位于血栓中心，纤维蛋白生成也是如此。本文提出的实验系统是一个有价值的工具，不仅用于研究血栓形成的机制，而且还评估血管系统内抗血栓药物的疗效。
The exact mechanism of blood vessel thrombus formation remains to be defined. Here, we introduce a new approach to probe thrombus formation in blood vessels of living animals using intravital microscopy in green fluorescent protein (GFP)-transgenic mice to simultaneously monitor platelet aggregation and procoagulant activity. To this end, GFP-expressing platelets and annexin A5 labeled with a fluorescent dye were employed to visualize and analyze platelet aggregation and markers of procoagulant activity (platelet surface phosphatidylserine (PS)). Laser-induced thrombi increased and then decreased in size with time in vessels of living animals, whereas platelet surface PS initiated at the site of injury and then penetrated into the thrombus. PS-positive platelets were predominantly localized in the center of the thrombus, as was fibrin generation. The experimental system proposed here is a valuable tool not only for investigating mechanisms of thrombus formation but also to assess the efficacy of antithrombotic drugs within the vasculature.