MOLECULAR GENETICS OF LIVER DEVELOPMENT

肝脏发育的分子遗传学

• 批准号: 6315455
• 负责人: DIDIER Y STAINIER
• 金额: $ 30.34万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6315455
• 关键词: developmental genetics; gene complementation; gene mutation; genetic mapping; genetically modified animals; histogenesis; liver; molecular genetics; nonmammalian vertebrate embryology; zebrafish

The liver performs several essential functions including the storage and filtration of blood, the secretion of bile, the synthesis and secretion of amino acids, and the conversions of sugars into glycogen. Despite these essential functions, little is known about the cellular and molecular mechanisms leading to the formation of the liver. We propose to use a forward genetic approach in zebrafish to investigate liver development. The gut tube ad its associated organs are not readily visible in the developing zebrafish embryo and thus have not been very accessible to morphological screens. Using a transgenic line which expresses GFP in the developing gut tube and its associated organs, the liver and pancreas, we plan to conduct a large-scale diploid screen for mutations that affect the formation of these organs. We will focus our future studies on mutations that affect the development of the liver. However, mutations that affect the formation of the pancreas and/or intestine will also be identified in our screen. We will set up a system to publish the phenotype of these mutations on the Web and make them available as soon as they are identified. In order to screen for mutations affecting the development of the liver, pancreas and intestine using this GFP transgenic line, we propose the following specific aims: 1) Characterize zebrafish liver development in detail. These studies will include further analysis of the GFP transgenic line as well as detailed examination of liver gene expression in wild-type and specific mutant backgrounds. 2) Screen for mutations that affect the formation of the liver, pancreas, and intestine. We will conduct a large- scale diploid screen using the GFP transgenic line. 3) Classify the liver mutations and begin to analyze several of them in detail. Complementation analysis, genetic mapping and molecular characterization will allow us to classify these mutations and select a few for further work. These molecular genetic studies of liver development will help provide the necessary context for further investigating the pathophysiology of malformations and malfunctions of the human liver, and may also facilitate differentiation-based strategies for treatment of a variety of diseased states.

肝脏执行几种基本功能，包括血液的储存和过滤，胆汁的分泌，氨基酸的合成和分泌以及糖转化为糖原。 尽管这些基本功能，但对导致肝脏形成的细胞和分子机制知之甚少。 我们建议在斑马鱼中使用远期遗传学方法来研究肝发育。肠管广告的相关器官在发育中的斑马鱼胚胎中不容易看到，因此形态学筛选不太容易进入。 我们使用在发育中的肠管及其相关器官，肝脏和胰腺中表达GFP的转基因线，我们计划对影响这些器官形成的突变进行大规模的二倍体筛选。 我们将将未来的研究集中在影响肝脏发展的突变上。 但是，影响胰腺和/或肠形成的突变也将在我们的屏幕上确定。 我们将建立一个系统，以在网络上发布这些突变的表型，并在确定后立即提供它们。 为了筛选使用该GFP转基因线影响肝脏，胰腺和肠发展的突变，我们提出以下特定目的：1）详细详细介绍斑马鱼肝发育。 这些研究将包括对GFP转基因线的进一步分析，以及对野生型和特定突变体背景中肝基因表达的详细研究。 2）筛选影响肝脏，胰腺和肠形成的突变。 我们将使用GFP转基因线进行大规模的二倍体屏幕。 3）对肝脏突变进行分类，并开始详细分析其中几个。互补分析，遗传图和分子表征将使我们能够对这些突变进行分类，并选择一些进行进一步工作。 这些对肝发育的分子遗传研究将有助于提供必要的背景，以进一步研究人肝脏的畸形和故障的病理生理学，并且还可能促进基于分化的策略来治疗各种患病状态。