Global Gene Expression Atlas of the Developing Kidney

描述（由申请人提供）：目的是创建发育中肾脏的全球基因表达图谱。中心论点是，激光捕获显微切割和微阵列的组合可以用来有效地实现这一目标。具有基本上完整的基因表达的微阵列可用于快速确定发育中肾脏的激光捕获显微切割元件中每个基因的表达水平。因此，一个单一的实验，提供了一个组件的基因表达状态的综合分析，和有限数量的实验检查每个结构和子结构可以创建一个图谱。具体目标1是使用这种策略来产生发育中的小鼠肾脏的特定结构域的基因表达谱图谱。最初的重点将是E15.5肾脏，它提供了一个单一的时间点与多个阶段的肾发生，但更早的时间点也将进行检查。将使用结构、凝集素染色、免疫组织化学和转基因GFP表达的组合来精确鉴定特定组分和谱系。具体目标2是使转基因小鼠工具，以促进基因表达谱和肾脏发育的功能研究。我们提出了一系列的转基因小鼠与特定的启动子驱动限制性表达的Cre-GFP盒。这些小鼠将用于双重目的，允许鉴定特异性目的1的额外离散肾脏组分，并有助于将来在发育中的肾脏中进行结构域特异性基因敲除研究。具体目标3是对微阵列数据进行生物信息学分析，并使研究社区随时可以获得结果。微阵列产生需要筛选的大量基因列表。对特定目标1中定义的基因表达的复杂编排的分析将提供对肾单位不同部分发育的遗传基础的更深入的了解。

DESCRIPTION (provided by applicant): The objective is to create a global gene expression atlas of the developing kidney. The central thesis is that a combination of laser capture microdissection and microarrays can be used to efficiently achieve this goal. Microarrays with essentially complete gene representation can be used to rapidly determine the expression levels of every gene in laser capture microdissected elements of the developing kidney. A single experiment, therefore, provides a comprehensive analysis of the gene expression status of one component, and a limited number of experiments examining each structure and substructure can create an atlas. Specific aim 1 is to use this strategy to produce an atlas of the gene expression profiles of specific domains of the developing mouse kidney. The initial focus will be on the E15.5 kidney, which provides a single time point with multiple stages of nephrogenesis, but earlier time points will also be examined. A combination of structure, lectin staining, immunohistochemistry and transgenic GFP expression will be used to precisely identify specific components and lineages. Specific aim 2 is to make transgenic mouse tools to promote both gene expression profiling and functional studies of kidney development. We propose to make a series of transgenic mice with specific promoters driving restricted expression of a Cre-GFP cassette. These mice will serve a dual purpose, to allow identification of additional discreet kidney components for Specific Aim 1 and to aid future domain specific gene knockout studies in the developing kidney. Specific aim 3 is to perform bioinformatics analysis of the microarray data and to make results readily available to the research community. Microarrays produce large gene lists that need to be sifted. Analysis of the complex orchestrations of gene expression defined in Specific Aim 1 will provide deeper insight into the genetic basis of the development of the distinct parts of the nephron.