In vivo real-time two-photon microscopic imaging of platelet aggregation induced by selective laser irradiation to the endothelium created in the beta-actin-green fluorescent protein transgenic mice
In vivo real-time two-photon microscopic imaging of platelet aggregation induced by selective laser irradiation to the endothelium created in the beta-actin-green fluorescent protein transgenic mice
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DOI：
10.1007/s11239-011-0600-y
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发表时间：
2011-08-01
影响因子：
4
通讯作者：
Kusunoki, Masato
中科院分区：
文献类型：
--
作者：
Koike, Yuhki;Tanaka, Koji;Kusunoki, Masato
尽管最近报道了体内血栓形成，但所有先前的激光诱导血栓模型都与血管壁破坏有关。本研究旨在评估选择性内皮损伤后体内实时血小板聚集，并可视化血栓形成，而不会破坏双光子激光照射引起的动脉内膜和肿胀。使用双光子激光扫描显微镜（TPLSM）通过选择性内皮损伤在β-肌动蛋白绿色荧光蛋白转基因小鼠中产生盲肠小动脉血栓。评估体内血栓形成的实时过程。还详细分析了抗血栓药物的功效。 TPLSM 可以实现从动脉平滑肌到内膜层的微血管成分的可视化。选择性激光照射内膜层后，在损伤区域下游形成线状粘连后，仅在内膜层损伤区域可见血小板粘附和聚集。当剪切应力被克服时，血栓在受伤区域的下游边缘开始形成。激光照射后约 60 分钟血栓体积趋于稳定。精确评估抗凝药物的溶栓效果；因此，我们的模型似乎是目前报道的体内病理生理过程实时成像最先进的模型。使用 TPLSM 与器官稳定系统相结合可以实现血栓形成的体内实时成像。 TPLSM 的高放大倍率和分辨率允许研究血栓形成机制以及评估抗血栓药物疗效，且实验间差异很小。
Although thrombus formation in vivo has recently been reported, all previous laser induced thrombus models have been associated with vessel wall disruption. This study aimed to evaluate in vivo real-time platelet aggregation after selective endothelial injury, and to visualize thrombus formation without disruption and swelling of the arterial intima induced by two-photon laser irradiation. Cecal arteriole thrombi were created in beta-actin-green fluorescent protein transgenic mice by selective endothelial injury using two-photon laser-scanning microscopy (TPLSM). The in vivo real-time process of thrombus formation was assessed. Anticoagulant drug efficiency for thrombi was also analyzed in detail. TPLSM allowed visualization of microvessel components from the arterial smooth muscle to the intimal layer. Immediately after selective laser irradiation of the intimal layer, platelet adhesion and aggregation were seen only at the area of injury of the intimal layer after forming linear adhesions downstream of the injured area. When shear stress was overcome, thrombus formation began at the downstream edge of the injured area. Thrombus volume plateaued approximately 60 min after laser irradiation. The thrombolytic effects of anticoagulant drugs were precisely assessed; therefore, our model appears the most advanced model in point of real-time imaging of pathophysiological processes in vivo currently reported. In vivo real-time imaging of thrombus formation can be achieved using TPLSM in combination with an organ stabilizing system. The high magnification and resolution of TPLSM allows investigation of the mechanisms of thrombus formation along with assessment of antithrombotic drug efficacy with little interexperimental variation.