Organogenesis Screen Using Tissue-Specific Zebrafish

描述：（申请人的摘要）器官形成需要许多步骤， 将来自不同来源的具有特定命运的细胞组装在一起， 单位如何使一个适当形成的器官并保持其正常功能是一个 复杂生物体的重要任务参与基因突变的基因 器官的模式和/或功能通常导致胚胎死亡，或 在以后生活中引起严重的健康问题，如先天性心脏病 衰竭心律不齐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。深入了解这些问题， 这将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。 为了了解脊椎动物器官是如何形成的，并确定基因和 参与脊椎动物器官发生的途径。我们就来 使用斑马鱼作为遗传学、胚胎学和基因组学方法的组合， 模型系统斑马鱼是脊椎动物的模式生物， 大规模基于表型遗传筛选。斑马鱼研究进展 基因组资源允许突变的快速作图和定位克隆 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特性使其适合 GFP技术的应用。我们将培育出一个转基因品系 其中6个器官原基被GFP表达标记。我们将会用这些 鱼来构建这些器官的四维命运图。这些信息 为进一步研究脊椎动物器官的形成奠定了基础。我们 将用ENU对这些鱼进行诱变，并筛选影响的突变 斑马鱼器官的形成和功能。我们还将开发试剂， 技术，以促进逆转录病毒插入功能获得性筛选。 从该屏幕识别的突变将提供解剖的切入点 参与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.