Automated Screening System for Modifiers of Cardiac Output in Zebrafish

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。已经进行小分子筛选以鉴定正常心血管过程的修饰剂和心血管疾病的修饰剂。正在开发的高通量系统旨在测量心率，但不解决心血管功能的其他关键参数，如血流速度，每搏输出量和射血分数。我们建议开发一种高通量系统，该系统将同时：1）测量心率; 2）生成腹主动脉的速度曲线; 3）测量舒张末期和收缩末期心室容积。基于显微镜的系统将利用多普勒光谱域光学相干断层扫描技术在几秒钟内生成斑马鱼胚胎和幼虫的三维结构图像，并测量血流。在第一阶段，我们开发了一个试验板系统，使收集的图像从背侧和腹侧的意见的动物。在试验板系统中，定位动物并手动调整扫描角度。在第II阶段，将开发一个完整的自动筛选系统，包括：1）软件算法，用于快速定位动物进行成像，并沿着其轴向长度调整扫描角度; 2）自动图像收集; 3）自动图像处理。第二阶段测试将包括一个试点屏幕，以检测心输出量减少的动物。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。特别是，这种筛选有望加速治疗心血管疾病的新药的发现。这些技术还将使心脏毒性的高通量测定成为可能。

DESCRIPTION (provided by applicant): The zebrafish, Danio rerio, has become widely accepted as a model for human disease. Small molecule screens have been performed to identify both modifiers of normal cardiovascular processes and modifiers of cardiovascular disease. High-throughput systems under development aim to measure heart rate but do not address other critical parameters of cardiovascular function such as blood velocity, stroke volume and ejection fraction. We propose to develop a high-throughput system that would simultaneously: 1) measure heart rate; 2) generate a velocity profile of the ventral aorta; and 3) measure end diastolic and end systolic ventricular volume. The microscope-based system will utilize spectral domain optical coherence tomography with Doppler to generate, within a matter of seconds, a three dimensional structural image of zebrafish embryos and larvae and measure blood flow. In Phase I we developed a breadboard system that enabled collection of images from both the dorsal and ventral views of the animals. In the breadboard system the animals were located and the scan angle was adjusted manually. In Phase II a complete, automated screening system will be developed that includes: 1) software algorithms to rapidly locate the animals for imaging and adjust the scan angle along their axial length; 2) automated image collection; and 3) automated image processing. Phase II testing will include a pilot screen to detect animals with decreased cardiac output. 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. In particular, such screens are expected to accelerate discovery of new pharmaceuticals to treat cardiovascular diseases. These technologies will also enable high-throughput assays of cardiotoxicity.