RENAL MICROVASCULAR ENDOTHELIAL CELL DIFFERENTIATION

肾小球和肾小管毛细血管单位是专门的过滤 具有复杂的基质组织和细胞-细胞相互作用的装置。 这些单位的功能障碍在各种肾脏疾病中被注意到。 虽然对肾小球足细胞和肾小管上皮细胞的了解很多， 上皮细胞发育，连接复合体形成，裂隙隔 形成，功能和对损伤的反应，对这些功能的了解要少得多。 肾小管上皮细胞的发育、维持和对损伤的反应， 肾小球内皮细胞（EC）。 总的来说， 发育和对损伤的反应被认为是调节的， 部分，由周围的组成和组织 细胞外基质（ECM）和可溶性因子在局部环境中的作用。 此外，EC与自身以及ECM的这种动态相互作用 被认为涉及几类细胞粘附分子（CAM） 例如PECAM-1，基质粘附分子（SAM），例如 整联蛋白和细胞连接相关分子（JAM）如ZO 1。 在这个建议中，我们将研究层次（时空） 这三个蛋白质家族的选定部分的相互关系 在微血管内皮细胞形成管的过程中， 体外以及在胎儿和新生大鼠肾脏中。 具体来说，我们将 研究PECAM-1在启动和调节过程中的作用 在体外和体内的血管生成。 的相互作用 PECAM-1的细胞质结构域与丝状元件， 皮层膜细胞骨架也将被研究。 我们将 利用重组蛋白的亲和层析;稳定 细胞质结构域的转染和过表达以及截短 内皮细胞中该PECAM-1结构域的突变体;功能性PECAM-1 抗体和可溶性PECAM-1抗原;并利用 免疫化学和免疫组织化学方法来评估 PECAM-1在内皮细胞上的定位和组织 面

The glomerular and tubular capillary units are specialized filtration apparati having complex matrix organization and cell-cell interactions. Dysfunction of these units is noted in a variety of renal diseases. While a good deal is known about glomerular podocyte and tubular epithelial cell development, junction complex formation, slit diaphragm formation, function and response to injury, much less is known about the development, maintenance and the response to injury of tubular and glomerular endothelial cells (EC). In general, EC dynamics during development and in response to injury are thought to be modulated, in part, by the composition and organization of the surrounding extracellular matrix (ECM) and soluble factors in the local environment. furthermore, this dynamic interaction of EC with themselves and with ECM is thought to involve several classes of cell adhesion molecules (CAMs) such as PECAM-1, substratum adhesion molecules (SAMs) such as the integrins and cell junctional associated molecules (JAMs) such as ZO1. In this proposal we will examine the hierarchical (spatiotemporal) interrelationships of selected moieties of these three protein families in the processes of tube formation by microvascular endothelial cells in vitro and in the fetal and newborn rat kidney. Specifically, we will examine the roles of PECAM-1 in initiating and modulating the processes of in vitro and in vivo angiogenesis. The interactions of the cytoplasmic domain of PECAM-1 with elements of the filamentous and cortical membrane cytoskeleton will also be investigated. We will utilize affinity chromatography with recombinant proteins; stable transfection and over-expression of the cytoplasmic domain and truncation mutants of this PECAM-1 domain in endothelial cells; functional PECAM-1 antibodies and soluble PECAM-1 antigen in inhibition studies; and utilize immunochemical and immunohistochemical methods to assess the localizations and organization of PECAM-1 on the endothelial cell surface.