Identification and characterization of zebrafish thrombocytes
Identification and characterization of zebrafish thrombocytes
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DOI：
10.1046/j.1365-2141.1999.01763.x
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发表时间：
1999-12-01
影响因子：
6.5
通讯作者：
Troyer, D
中科院分区：
文献类型：
--
作者：
Jagadeeswaran, P;Sheehan, JP;Troyer, D
为了分析斑马鱼的原发性止血，我们通过形态学、免疫学和功能学方法鉴定并表征了斑马鱼血小板。开发了新的方法用于收获斑马鱼血液，保存血小板，并在微量滴定板中测定全血粘附/聚集反应。光和电子显微镜的血小板示出的形态特征，包括形成的聚集体，伪足，和表面连接的囊泡analagous的血小板小管系统。用多克隆抗血清对人血小板糖蛋白的免疫染色证明血小板表面存在糖蛋白Ib和IIb/IIIa样复合物。粘附/聚集和ATP释放的全血测定显示，瑞斯托霉素诱导的粘附无ATP释放，血小板激动剂（胶原、花生四烯酸）诱导的聚集有ATP释放。从用阿司匹林处理的斑马鱼收获的血液表现出对花生四烯酸诱导的聚集和激动剂诱导的ATP释放的抑制，这与至少部分依赖于完整的环加氧酶途径一致。结合形态学、免疫学和功能学证据表明，斑马鱼血小板是哺乳动物血小板的止血同源物。保护血小板功能和凝血的主要止血途径表明，斑马鱼是哺乳动物止血和血栓形成的相关模型。
To analyse primary haemostasis in the zebrafish we have identified and characterized the zebrafish thrombocyte by morphologic, immunologic and functional approaches. Novel methods were developed for harvesting zebrafish blood with preservation of thrombocytes, and assaying whole blood adhesion/aggregation responses in microtitre plates. Light and electron microscopy of the thrombocyte illustrated morphological characteristics including the formation of aggregates, pseudopodia, and surface-connected vesicles analagous to the platelet canalicular system. Immunostaining with polyclonal antisera versus human platelet glycoproteins demonstrated the presence of glycoprotein Ib and IIb/IIIa-like complexes on the thrombocyte surface. Whole blood assays for adhesion/aggregation and ATP release showed ristocetin-induced adhesion without ATP release, and platelet agonist (collagen, arachidonic acid) induced aggregation with ATP release. Blood harvested from zebrafish treated with aspirin demonstrated inhibition of arachidonic acid induced aggregation and agonist induced ATP release, consistent with at least partial dependence on an intact cycle oxygenase pathway. The combined morphologic immunologic and functional evidence suggest that the zebrafish thrombocyte is the haemostatic homologue of the mammalian platelet. Conservation of major haemostatic pathways involved in platelet function and coagulation suggests that the zebrafish is a relevant model for mammalian haemostasis and thrombosis.