Novel System to Automate Zebrafish Angiogenesis Assays

自动化斑马鱼血管生成检测的新系统

• 批准号: 6994544
• 负责人: ANTHONY A Ferrante
• 金额: $ 12.97万
• 依托单位: PHYSICAL SCIENCES, INC
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-14 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6994544
• 关键词: angiogenesis; angiogenesis inhibitors; bioimaging /biomedical imaging; biomedical automation; biotechnology; cardiovascular imaging /visualization; drug discovery /isolation; drug screening /evaluation; genetically modified animals; green fluorescent proteins; high throughput technology; technology /technique development; zebrafish

DESCRIPTION (provided by applicant): The zebrafish, Danio rerio, has become widely accepted as a model for human disease. There is strong interest in developing zebrafish as a model organism for use in high throughput drug discovery assays. Advantages of using zebrafish in drug discovery include the ease with which they absorb candidate drug molecules and their genetic and physiologic similarity to humans. Thus far, high throughput screening in zebrafish has been limited to embryos and very young larvae. This is because the vasculature and many of the organs that are most interesting to researchers are best viewed from the side of the animal. Once the swim bladder inflates the larvae generally float upright making visualization of internal structures difficult. This is especially true in the multiwell microplate formats that are used in most high-throughput drug discovery. Efforts to resolve this important problem have centered on use of confocal imaging techniques or powerful image deconvolution software to assemble 3-D reconstructions of the animal. These efforts have not achieved the desired spatial resolution and typically require 1 to 2 minutes to image a single well. Physical Sciences Inc. (PSI) has invented a novel technology that will be adaptable to standard image analysis systems that are already in use in drug discovery laboratories. PSI's technology will allow researchers to generate clear side-views of older zebrafish larvae in microwell arrays. By providing a side view of the zebrafish, the technology will enable generation of quality images in seconds per well versus minutes per well for current techniques, hi Phase I we will generate fluorescence images of the vasculature of transgenic zebrafish that express Enhanced Green Fluorescent Protein (EGFP) under control of thefli-1 regulatory sequences. We will compare images of larvae with normal vascular development with those of larvae treated with a VEGF inhibitor and which have incompletely developed vasculature. These experiments will serve as a model for Phase II development of a fully-automated, high-throughput, drug-discovery assay in zebrafish for novel angiogenesis inhibitors.

描述（由申请人提供）： 斑马鱼Danio Rerio已被广泛接受为人类疾病的模型。人们对开发斑马鱼作为模型生物有浓厚的兴趣，以用于高吞吐药物发现分析。在药物发现中使用斑马鱼的优点包括它们吸收候选药物分子的容易性以及与人类的遗传和生理相似性。迄今为止，斑马鱼中的高吞吐量筛查仅限于胚胎和非常年轻的幼虫。这是因为脉管系统和许多最有趣的研究人员的器官最好从动物的侧面观看。一旦游泳膀胱膨胀，幼虫通常会直立地漂浮，从而使内部结构的可视化变得困难。在大多数高通量药物发现中使用的多醇微板格式中尤其如此。解决这一重要问题的努力集中在使用共聚焦成像技术或强大的图像反卷积软件来组装动物的3-D重建方面。这些努力尚未达到所需的空间分辨率，通常需要1到2分钟才能成像一个孔。物理科学公司（PSI）发明了一种新型技术，该技术将适应已经在药物发现实验室中使用的标准图像分析系统。 PSI的技术将使研究人员能够在Microwell阵列中生成清晰的斑马鱼幼虫的清晰侧视图。通过提供斑马鱼的侧视图，该技术将在当前技术中以每井和每孔的数分井的几秒钟而产生高质量的图像。我们将将幼虫的图像与正常的血管发育与用VEGF抑制剂治疗的幼虫发育的图像进行比较，而幼虫的图像并未完全开发脉管系统。这些实验将作为用于新型血管生成抑制剂中全自动，高通量，药物发现测定法的II期开发模型。