Organogenesis Screen Using Tissue-Specific Zebrafish

描述：(申请者摘要)器官形成需要许多步骤才能 将来自不同来源、具有特定命运的细胞组装成一个功能 单位。如何制造一个结构合理的器官并保持其正常功能是一个 对于复杂的生物来说，这是一项至关重要的任务。基因的异常涉及到 器官的图案和/或功能通常会导致胚胎死亡，或者 在以后的生活中引发严重的健康问题，如先天性心脏病 失败、心律失常、白血病和糖尿病。了解器官是如何 模式化，哪些基因参与器官模式化和/或功能，以及 这些基因如何相互作用是研究的重要问题。 脊椎动物器官的形成和功能。对这些问题的深入了解不要 只会有助于进一步研究器官发生的基本机制， 而且还将导致早期诊断和治疗的发展 人类疾病。 了解脊椎动物器官是如何形成的，并识别基因和 参与脊椎动物器官发生的途径。我们将采取一项 以斑马鱼为研究对象的遗传学、胚胎学和基因组学方法的结合 模型系统。斑马鱼是脊椎动物模式生物，可以促进 大规模的基于表型的遗传筛选。斑马鱼研究进展 基因组资源允许对突变的人进行快速定位和位置克隆 基因。此外，斑马鱼的器官系统是所有器官系统中最简单的形式 脊椎动物。斑马鱼胚胎的透明性质使其适合 为绿色荧光技术的应用。我们将产生一株转基因株系 其中6个器官原基以绿色荧光蛋白的表达为标志。我们将使用这些 鱼来构建这些器官的4维命运图。此信息 将为今后脊椎动物器官形成的研究提供基础。我们 将用ENU诱变这些鱼并筛选影响 斑马鱼器官的形成和功能。我们还将开发试剂和 促进逆转录病毒插入功能增益筛查的技术。 从这一筛选中发现的突变将为解剖提供切入点 脊椎动物器官发生的遗传和分子途径。

DESCRIPTION: (Applicant's Abstract)Organ formation requires many steps to assemble cells from different origins with specific fates to form a functional unit. How to make a properly formed organ and maintain its normal function is a vital task for complex organisms. Abnormalities in genes that are involved in the patterning and/or function of organs often cause embryonic lethality, or induce severe health problems later in life, as evident in congenital heart failure, arrhythmia, leukemia and diabetes. Understanding how the organs are patterned, which genes are involved in organ patterning and/or function, and how these genes interact with each other are important issues for studying vertebrate organ formation and function. In-depth knowledge of these issues not only will facilitate further studies on the basic mechanisms of organogenesis, but also will lead to the development of early diagnosis and treatment for human diseases. To understand how vertebrate organs are formed and to identify genes and pathways that are involved in vertebrate organogenesis. We will take a combination of genetic, embryological and genomic approach using zebrafish as a model system. Zebrafish is the vertebrate model organism that can facilitate large-scale phenotype-based genetic screen. Recent advances in zebrafish genomic resources allow rapid mapping and positional cloning of the mutated genes. Furthermore, the zebrafish organ systems are the simplest forms of all vertebrates. The transparent nature of the zebrafish embryos makes it suitable for the application of the GFP technology. We will generate a transgenic line in which six organ primordia are marked by GFP expression. We will use these fish to construct a 4-dimensional fate map for these organs. This information will provide foundation for future studies of vertebrate organ formation. We will mutagenize these fish with ENU and screen for mutations affecting zebrafish organ formation and function. We will also develop reagents and technologies to facilitate a retroviral insertional gain-of-function screen. Mutations identified from this screen will provide entry points to dissect genetic and molecular pathways involved in vertebrate organogenesis.