In Vivo Zebrafish Tyrosine Kinase ELISA

描述（由申请人提供）：小分子ATP竞争性激酶抑制剂（如格列卫、易瑞沙和特罗凯）最近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靶点之一。由于它们与人类的遗传和生理相似性，斑马鱼显示出作为药物筛选的有效动物模型的希望。虽然哺乳动物蛋白激酶的所有成员尚未在斑马鱼中被鉴定，但原位杂交实验表明Src家族的酪氨酸激酶src、fyn和yes在斑马鱼胚胎中普遍表达。在本研究中，我们将开发一种高通量的斑马鱼体内ELISA方法来鉴定Src家族激酶抑制剂。斑马鱼测定允许使用类似于基于细胞的测定的通量来评估化合物在复杂生理环境中的治疗潜力。斑马鱼测定的优点包括：测定时间短、成本低、在药物递送期间易于胚胎维持、测定程序与标准微孔板形式的兼容性、每个胚胎的单一药物剂量以及测试大量胚胎以获得统计学显著结果的能力。公共卫生相关性：在本研究中，我们将开发一种高通量的斑马鱼体内ELISA方法来鉴定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.