PODOCYTE CELL LINEAGE IN GENITOURINARY DEVELOPMENT

描述（改编自应用程序） 申请人的研究计划的长期目标将测试以下内容 两种假设：1）足细胞（肾小球内脏上皮细胞）是 所需的肾小球血管生成在发展过程中和2）， 足细胞在慢性肾功能不全的进展中起关键作用， 各种潜在原因，包括糖尿病，衰老和炎症。的 已经克隆了足细胞特异性基因nephrin的启动子。转基因 含有1.25-kb的人nephrin基因的5'侧翼区 概括了nephrin mRNA转录本的内源性表达， 仅在毛细血管袢和成熟期肾小球足细胞中表达， 发育中的鼠肾拟议的研究将使用这种启动子， 产生新的试剂来描述足细胞在肾脏中的作用 发展和疾病。本申请的主要目的是 开发小鼠模型系统以在体外操纵足细胞基因表达， in vivo.具体目标包括：1.为了产生足细胞特异性的 Cre-重组酶小鼠品系，以便在一个 细胞特异性的方式在肾脏发育。2.生成鼠标创始人 可以用于以诱导方式修饰基因活性的品系， 足细胞3.为了分离新的小鼠足细胞系， “活”的分化标志。4.为了进一步表征顺式作用 nephrin基因的1.25-kb足细胞特异性启动子内的元件 为了确定负责的反式作用因素， 足细胞特异性表达。5.为了产生足细胞特异性敲除 血管内皮生长因子（VEGF）。 在本申请中产生的试剂将允许时间和时间上的测量。 足细胞中特异性基因活性的空间修饰。将 可能操纵参与肾小球血管生成的候选基因， 以确定足细胞在建立肾小球滤过中的作用 屏障在未来的研究中，这些试剂将有助于确定 肾小球硬化症中足细胞的数量，并最终确定治疗 慢性肾病的治疗方法

DESCRIPTION (adapted from the application) The long-term goals of the applicant's research program will test the following two hypotheses: 1) that the podocyte (glomerular visceral epithelial cell) is required for glomerular angiogenesis during development and 2) that the podocyte is pivotal in progression of chronic renal dysfunction due to a variety of underlying causes that include diabetes, aging and inflammation. The promoter of a podocyte-specific gene, nephrin, has been cloned. A transgene containing 1.25-kb of the 5' flanking region of the human nephrin gene recapitulates the endogenous expression of the nephrin mRNA transcript, and is only expressed in podocytes in capillary-loop and mature stage glomeruli of the developing murine kidney. The proposed studies will use this promoter to generate novel reagents to delineate the role of the podocyte in kidney development and disease. The main objective of the present application is to develop mouse model systems to manipulate podocyte gene expression in vitro and in vivo. The Specific Aims include: 1. To generate a podocyte-specific Cre-recombinase mouse line in order to delete and over-express genes in a cell-specific manner during renal development. 2. To generate mouse founder lines that can be used to modify gene activity in an inducible manner in podocytes. 3. To isolate novel murine podocyte cell lines that expresses a "living" marker of differentiation. 4. To further characterize the cis-acting elements within the 1.25-kb podocyte-specific promoter of the nephrin gene in order to identify trans-acting factors that are responsible for podocyte-specific expression. 5. To generate a podocyte-specific knockout of vascular endothelial growth factor (VEGF). The reagents generated in the present application will allow both temporal and spatial modification of gene activity specifically in podocytes. It will be possible to manipulate candidate genes involved in glomerular angiogenesis and to determine the role of the podocyte in setting up the glomerular filtration barrier. In future studies, these reagents will be useful to determine the role of the podocyte in glomerulosclerosis and ultimately, to identify therapeutic targets for chronic renal disease.