Transgenic zebrafish for neurobiology

描述(申请人提供)：斑马鱼已经成为研究神经发育、生理和疾病的强大模型系统。它是这一特殊目的的理想选择，因为它是一种小而半透明的脊椎动物，具有广泛的行为能力。尽管取得了这一进展，但能够充分利用光遗传方法的转基因品系仍然相对较少。例如，该领域仍然缺乏针对大多数神经递质或多肽的特定驱动因素和记者，现有的标记特定大脑区域或核团的线条往往缺乏必要的特异性。因此，斑马鱼作为一个模式系统仍然缺乏果蝇社区在过去十年中建立的基本工具集。为了克服这些限制，我们建议建立转基因品系，允许在由大脑区域和/或神经递质亚型指定的受限神经元亚群中特定表达感兴趣的蛋白质。我们将使用增强子陷阱产生240条线，通过修改的Gal4载体(目标1)驱动区域和细胞类型的特定神经表达，我们将使用基因组工程产生12条线，驱动产生主要类型的神经递质的神经元表达(目标2)。该企业的一个重要组成部分将是设计和建立一个在线界面，提供每一种转基因品系的细节。研究人员将获得每条直线特征的广泛细节和注释良好的图像数据集。Engert和Schier实验室在斑马鱼神经系统的解剖和功能成像以及拟议项目所需的分子和遗传工具箱方面拥有丰富的专业知识。该项目将得到由斑马鱼神经生物学领域领导人组成的咨询委员会的进一步支持。

DESCRIPTION (provided by applicant): The zebra fish has emerged as a powerful model system to study neural development, physiology, and disease. It is ideal for this particular purpose since it is a small and translucent vertebrate with a wide behavioral repertoire. Despite this progress, there are still comparatively few transgenic lines that allow the full exploitation f optogenetic approaches. For example, the field still lacks specific drivers and reporters for most neurotransmitters or peptides, and existing lines that mark specific brain regions or nuclei often lack the necessary specificity. The zebra fish as a model system is thus still lacking the essential tool set that the Drosophila community has built in the last decade. To overcome these limitations, we propose to generate transgenic lines that allow the specific expression of proteins of interest in restricted subpopulations of neurons that are specified by brain region and/or by neurotransmitter subtype. We will use enhancer trapping to generate 240 lines that drive region- and cell- type specific neural expression via a modified Gal4 vector (Aim 1), and we will use genome engineering to generate 12 lines that drive expression in neurons producing the major classes of neurotransmitters (Aim 2). An essential component of the enterprise will be the design and establishment of an online interface that will provide details of each transgenic line. Researchers will be provided with extensive details of each line's characteristics and well-annotated image datasets. The Engert and Schier lab have extensive expertise in anatomical and functional imaging of the zebra fish nervous system as well as the molecular and genetic toolbox that is necessary for the proposed project. The project will be further supported by an advisory committee consisting of leaders in the field of zebra fish neurobiology.