In vivo imaging analysis of the interaction between unusually large von Willebrand factor multimers and platelets on the surface of vascular wall
In vivo imaging analysis of the interaction between unusually large von Willebrand factor multimers and platelets on the surface of vascular wall
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DOI：
10.1007/s00424-011-0958-x
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发表时间：
2011-06-01
影响因子：
4.5
通讯作者：
Urano, Tetsumei
中科院分区：
文献类型：
--
作者：
Rybaltowski, Miroslaw;Suzuki, Yuko;Urano, Tetsumei
为了阐明异常大的冯维勒布兰德因子 (UL-VWF) 多聚体如何促进血栓形成，我们对它们的行为以及缺乏编码裂解 UL-VWF 的蛋白酶基因的活体小鼠的血小板行为进行了分析，UL-VWF 是一种具有 1 型血小板反应蛋白基序 13 (ADAMTS13-/-) 的解整合素样金属蛋白酶。将ADAMTS13-/-小鼠与表达绿色荧光蛋白的转基因小鼠(GFP小鼠)杂交，获得GFP-ADAMTS13-/-小鼠。采用活体共聚焦荧光显微镜监测表达 GFP 的血小板的动态。施用加压素衍生物 DDAVP（一种 VWF 促分泌剂）可增加粘附在肠系膜上抗 VWF 抗体识别位点的血管内皮细胞 (VEC) 上的血小板数量。其中一些血小板相互连接并排列成串上的珠子。它们在 5 分钟时达到最大长度，并且在 GFP-ADAMTS13-/- 小鼠中比在 GFP 小鼠中更长（5.3 +/- 4.3，N=6 vs 2.9 +/- 2.1 μm，N=4）（平均值+/- SE）。局部应用 FeCl(3) 造成的 VEC 局灶损伤时间更长（25、3-50 vs 10、2-25 μ m，P
To elucidate how unusually large von Willebrand factor (UL-VWF) multimers facilitate thrombus formation, their behavior was analyzed together with that of platelets in living mice deficient in the gene encoding the protease that cleaves UL-VWF, a disintegrin-like and metalloprotease with thrombospondin type 1 motif 13 (ADAMTS13-/-). By crossing ADAMTS13-/- mice with green fluorescent protein-expressing transgenic mice (GFP mice), GFP-ADAMTS13-/- mice were obtained. The dynamics of GFP-expressing platelets were monitored employing intravital confocal fluorescent microscopy. Administration of a vasopressin derivative, DDAVP, a secretagogue of VWF increased the number of platelets adhered to vascular endothelial cells (VECs) on mesentery at sites recognized by an anti-VWF antibody. Some of these platelets were interconnected and aligned as beads on a string. They reached their maximum length at 5 min and were longer in GFP-ADAMTS13-/- mice than in GFP mice (5.3 +/- 4.3, N=6 vs 2.9 +/- 2.1 mu m, N=4) (mean +/- SE). Focal injury of VECs by topical application of FeCl(3) developed longer (25, 3-50 vs 10, 2-25 mu m, P