In Vivo Zebrafish Tyrosine Kinase ELISA

描述（由申请人提供）：最近小分子atp竞争性激酶抑制剂（如Gleevec， Iressa和Tarceva）的成功引起了极大的关注，蛋白激酶现在代表了药物发现和开发的最重要的靶标组之一。由于斑马鱼在遗传和生理上与人类相似，因此有望成为药物筛选的有效动物模型。尽管哺乳动物蛋白激酶的所有成员尚未在斑马鱼中被鉴定出来，但原位杂交实验表明，酪氨酸激酶Src家族Src、fyn和yes在斑马鱼胚胎中普遍表达。在这项研究中，我们将开发一种高通量的斑马鱼体内ELISA格式来鉴定Src家族激酶抑制剂。斑马鱼试验允许在复杂的生理环境中使用类似于基于细胞的测定的吞吐量来评估化合物的治疗潜力。斑马鱼试验的优点包括：试验时间短，成本低，在给药过程中易于胚胎维护，试验程序与标准微孔板格式兼容，每个胚胎使用单一药物剂量，以及能够测试大量胚胎以获得统计显着结果。公共卫生相关性：在这项研究中，我们将开发一种高通量的斑马鱼体内ELISA格式，用于鉴定Src家族激酶抑制剂。

DESCRIPTION (provided by applicant): The recent success of small-molecule ATP-competitive kinase inhibitors such as Gleevec, Iressa, and Tarceva has attracted significant attention and protein kinases now represent one of the most important groups of targets for drug discovery and development. Because of their genetic and physiological similarity to humans, zebrafish show promise as an efficient animal model for drug screening. Although all members of the mammalian protein kinases have not yet been identified in zebrafish, in situ hybridization experiments showed that Src family of tyrosine kinases src, fyn and yes are ubiquitously expressed in zebrafish embryos. In this research, we will develop a high throughput in vivo zebrafish ELISA format for identifying Src family kinase inhibitors. The zebrafish assay permits evaluation of the therapeutic potential of a compound in a complex physiological environment using throughput similar to cell-based assays. Advantages of the zebrafish assay include: short assay time, low cost, ease of embryo maintenance during drug delivery, compatibility of assay procedures with standard microplate formats, single drug dosing for each embryo, and the ability to test large numbers of embryos for statistically significant results. PUBLIC HEALTH RELEVANCE: In this research, we will develop a high throughput in vivo zebrafish ELISA format for identifying Src family kinase inhibitors.