Jagadeeswaran Pudur, Paris Ryan, Rao Prashanth
Department of Cellular and Structural Biology, The University of Texas Health Science Center at San Antonio, USA.
Methods Mol Med. 2006;129:187-95. doi: 10.1385/1-59745-213-0:187.
Classical genetic approaches to study hemostasis and thrombosis have not been available until our recent introduction of the teleost, Danio rerio (the zebrafish), as an effective genetic model for in vivo coagulation assays. The genetic screen for this model is carried out using the genome saturation mutagenesis approach. The resulting mutants are screened for hemostatic or thrombotic defects. We developed a global physiological screening method for thrombosis by utilizing a laser to induce thrombosis in a specifically targeted area of the major artery and vein. Using this assay, we have screened many fish for abnormal hemostasis, and have isolated a number of mutants with abnormal coagulation parameters. These mutants can be grown, bred, and further evaluated for the genetic etiology of their abnormal hemostatic pathways.
直到我们最近引入硬骨鱼——斑马鱼(Danio rerio)作为体内凝血测定的有效遗传模型,才出现用于研究止血和血栓形成的经典遗传方法。该模型的遗传筛选是使用基因组饱和诱变方法进行的。对产生的突变体进行止血或血栓形成缺陷的筛选。我们开发了一种用于血栓形成的整体生理筛选方法,通过利用激光在大动脉和静脉的特定目标区域诱导血栓形成。使用该测定方法,我们筛选了许多鱼的止血异常情况,并分离出了一些具有异常凝血参数的突变体。这些突变体可以生长、繁殖,并进一步评估其异常止血途径的遗传病因。