Novel System for Long-Term Rearing and High-Throughput Assay of Zebrafish

描述（由申请人提供）：这项拟议研究的总体目标是开发一种多孔插入装置，该装置将有助于从胚胎到幼鱼发育阶段的活鱼中报告基因的快速自动检测。该系统将与利用荧光或发光报告基因作为评价感兴趣的生物学问题的手段的任何基于斑马鱼的测定相关。斑马鱼正在成为了解人类疾病和发现新的治疗化合物的重要模型。特别是，斑马鱼非常适合大规模筛选程序，提供了一个接近高通量能力的体内验证系统。充分实现这一潜力将需要开发自动化检测平台，例如利用荧光或发光读数的基于报告的系统，并包括处于更成熟发育阶段的鱼类。在第一阶段，将开发一个系统的组件，使胚胎能够在单个腔室中维持长达1个月的发育;并实现高通量的初步筛选。在第二阶段，将开发基于PSI SideView成像技术的系统的第二个组件，这将实现更严格的基于图像的辅助屏幕。 公共卫生相关性：这项研究将开发技术，使人类疾病的斑马鱼模型能够用于活性药物化合物的高通量筛选。这项技术将受益的一个关键领域是再生医学，特别关注用于糖尿病治疗的胰岛素产生胰腺细胞的再生。

DESCRIPTION (provided by applicant): The overall goal of this proposed research is to develop a multi-well insert apparatus that will facilitate rapid automated detection of reporter genes in living fish from embryonic to juvenile stages of development. This system will be relevant for any zebrafish-based assay which utilizes fluorescent or luminescent reporter genes as a means of evaluating a biological question of interest. Zebrafish are becoming an important model for understanding human disease and for discovery of new therapeutic compounds. In particular, zebrafish are well-suited to large-scale screening procedures, providing an in vivo validation system that approaches high-throughput capacities. Full realization of this potential will require the development of automated assay platforms, such as reporter based systems utilizing fluorescent or luminescent read outs, and the inclusion of fish at more mature stages of development. During Phase I, components of a system will be developed that will enable embryos to be maintained in individual chambers for up to 1 month of development; and enable a high-throughput primary screen. During Phase II a second component of the system based on the PSI SideView imaging technology will be developed that will enable a more rigorous, image-based secondary screen. PUBLIC HEALTH RELEVANCE: This research will develop technologies that will enable the use of zebrafish models of human disease in high-throughput screens for active pharmaceutical compounds. One key area that this technology will benefit is regenerative medicine with a particular focus in this application on regeneration of insulin-producing pancreatic cells for diabetes treatment.