Global Gene Expression Atlas of the Developing Kidney

描述（由申请人提供）：目标是创建发育中的肾脏的全球基因表达图谱。中心论点是激光捕获显微解剖和微阵列的结合可以有效地实现这一目标。具有基本完整基因表达的微阵列可用于快速确定激光捕获发育中的肾脏微解剖元素中每个基因的表达水平。因此，一个单一的实验，提供了一个成分的基因表达状态的全面分析，和有限数量的实验检查每个结构和亚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特定区域的基因表达谱图谱。最初的重点将放在E15.5肾脏上，它提供了一个单一的时间点，有多个肾脏形成阶段，但更早的时间点也将被检查。将结合结构、凝集素染色、免疫组织化学和转基因GFP表达来精确鉴定特定成分和谱系。具体目标2是使转基因小鼠成为促进肾脏发育的基因表达谱和功能研究的工具。我们打算制造一系列具有特定启动子驱动Cre-GFP盒式限制性表达的转基因小鼠。这些小鼠将有双重目的，一是为Specific Aim 1鉴定额外的肾脏成分，二是帮助未来在发育中的肾脏中进行区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，并使结果随时可供研究界使用。微阵列产生大量需要筛选的基因列表。对Specific Aim 1中定义的复杂基因表达编排的分析将为肾元不同部分发育的遗传基础提供更深入的了解。

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.