An automated in vivo screening platform for the UK zebrafish research community

我们对基因和药物功能的了解大多来自于所谓的筛选。筛选是检测基因和化合物功能的公正方法。例如，科学家可以测试大量基因的破坏如何影响感兴趣的生物过程，以发现哪个基因控制给定的过程。类似地，人们可以测试大量化合物如何影响特定的生物事件，并发现，例如，未来的药物如何影响不同的细胞。在活体动物身上进行筛选是非常困难的，因为人们无法在时间、成本或道德上合理的情况下，在足够多的动物身上操纵基因或使用化学品。然而，斑马鱼是一种实验室生物，可以很容易地进行遗传和化学筛选。这是因为它们的胚胎数量非常多（每天1000个），它们发育得非常快，在3天内从受精卵变成一个拥有人类大部分器官的微小实体——在技术上它们被认为是动物之前。这些年轻的生物体也是透明的，因此人们可以直接观察到随着时间的推移发生的生物事件。新的基因技术进一步完善了在活斑马鱼中可视化的能力，例如，通过创造出感兴趣的细胞类型荧光的鱼株。斑马鱼的基因和化学筛选已经教会了我们很多关于生命的基本分子和细胞机制的知识，甚至还导致了新的疾病治疗临床试验。然而，尽管有可能，这种筛选仍然是非常费力的手工程序，需要很长时间，而且往往依赖于直接实验者将基因或化学处理与生物学联系起来的能力。因此，非常需要提高这种筛查的效率，因为这将直接转化为对健康具有重要意义的新知识和见解。我们的方案结合了两个要素，一个是电子控制系统，它可以通过细小的毛细血管自动地将大量的斑马鱼胚胎从培养皿中一个接一个地快速而均匀地转移到显微镜架上。第二个元素是显微镜本身，它可以在单个斑马鱼通过系统时对其进行高速成像。人们可以拍摄整个动物的概况快照，以评估其总体健康状况或器官功能，或者对感兴趣的特定细胞进行非常详细的3D图像。因此，可以使用该系统在一天内系统地对数百条斑马鱼进行自动成像和筛选，并创建数据的永久记录，所有这些都代表了当前方法效率的巨大提高。这个新系统将被安装在爱丁堡大学，那里聚集了许多使用斑马鱼作为实验模型的世界专家。在资助期内，申请人将在爱丁堡安装该系统，并向更广泛的社区推广。在爱丁堡，该系统将支持专注于了解神经系统中运动神经元和神经胶质细胞发育的项目——这些细胞类型对正常的神经系统功能很重要，在运动神经元疾病和多发性硬化症等疾病中受到破坏。该系统还将用于获得色素细胞发育以及这些细胞如何在黑色素瘤中转化的新见解。进一步的筛选研究将进行，以更好地了解我们的免疫系统有时如何促进肿瘤生长，并了解我们大脑中免疫细胞的常驻网络如何发育和对损伤作出反应。更广泛的英国斑马鱼研究人员已经表达了极大的兴趣，他们将利用这项新技术来阐明生物学的进一步方面。因此，这种新的斑马鱼自动筛选系统将大大加强英国生物科学的健康。

Much of what we know about the function of genes and drugs has arisen from so-called screens. Screens are unbiased methods to test gene and chemical compound function. For example, scientists can test how disruption of large numbers of genes affects a biological process of interest to discover which gene controls a given process. Similarly one can test how large numbers of chemical compounds affect specific biological events and discover, for example, how future drugs might affect different cells. Carrying out screens on living animals is very difficult, because one cannot manipulate genes or use chemicals in a sufficiently large number of animals in a time, cost, or ethically reasonable manner. Zebrafish, however, are a laboratory organism in which genetic and chemical screens can be carried out readily. This is because their embryos are available in very large numbers (1000s per day), they develop very rapidly, going from a fertilized egg to a tiny entity with the majority of organs that humans have in under 3 days- before they are technically considered animals. These young organisms are also transparent, such that one can directly observe biological events as they happen over time. New genetic technologies have further refined the ability to visualize in living zebrafish, e.g. by creating fish strains in which cell types of interest fluoresce. Genetic and chemical screens in zebrafish have already taught us a great deal about the fundamental molecular and cellular mechanisms of life and have even led to new clinical trials for the treatment of disease. However, despite being possible, such screens have remained very laborious, manual procedures, that take extended periods of time and that often rely on immediate experimenter's ability to link a gene or chemical treatment with biology. Therefore there is a great need to increase the efficiency of such screens because this will translate directly into new knowledge and insights of importance to health. Our proposal combines two elements, an electronically controlled system that can automatically transfer large numbers of embryonic zebrafish from their dishes via small capillaries onto a microscope stand in a rapid and uniform manner one by one. The second element is the microscope itself, which can carry out high-speed imaging of the individual zebrafish as they pass through the system. One can take overview snapshots of the entire animal to assess general health or organ function, or very detailed 3D images of specific cells of interest. Therefore it is possible to use this system to automatically image and screen hundreds of zebrafish throughout a day in a systematic manner and create a permanent record of the data, all of which represents an enormous increase in the efficiency of current approaches.This new system will be installed at the University of Edinburgh, where there is a concentration of world-experts in the use of zebrafish as a laboratory model. During the funding period of this award, the applicants will install the system in Edinburgh, and launch it to the wider community. In Edinburgh, the system will support projects focused on understanding the development of motor neurons and glial cells in the nervous system- cell types that are important for normal nervous system function and disrupted in diseases such as motor neuron disease and MS. The system will also be used to gain new insights into pigment cell development and how these cells are transformed in melanoma. Further screening studies will be carried out to better understand how our immune system can sometimes promote tumour growth and to learn how the resident network of immune cells in our brain develops and responds to injury. The wider UK community of zebrafish researchers who have already expressed great interest will use this new technology to elucidate yet further aspects of biology. Thus this new automated screening system for zebrafish will greatly strengthen UK Bioscience for health.