MODEL FOR EPITHELIAL/BASEMENT MEMBRANE INTERACTIONS

细胞外基质（或其 特殊形式的基底膜）导致广泛的问题 与发育异常和继发性并发症有关， 糖尿病等疾病，基底膜增厚， 观察到功能障碍。我们对细胞机制的理解 正常和异常基底膜形成的基础和作用 在开发过程中， 由于缺乏适当的体外和体内模型， 实验方法解决这些问题。为了更好地接近这些 问题，我们已经开发了一种体内细胞模型，其中基底 膜形成可以通过实验在上皮细胞之间诱导， 双层及其形成与1）细胞分化过程有关 和形态发生，以及2）导致异常的疾病过程， 基底膜的结构和功能。该模型 由无脊椎动物水螅进化而来， 退化为上皮双层，其间有基底膜 （mesoglea）。基于先前发表的研究，我们定义了 实验条件，其中1）从分离的细胞产生的沉淀 从成年水螅形成上皮双层，存款， 基底膜，随后改革一个成年水螅结构内 72-96小时通过细胞分化和形态发生的过程， 2)与糖尿病有关， 高浓度葡萄糖下的细胞聚集-形态发生系统 在72-96小时内导致基底膜异常增厚 与哺乳动物所需的数月或数年相反的时间范围 糖尿病模型利用这种体内上皮/基底膜 模型系统，我们建议测试假设针对细胞 基底正常和异常发育的机制 膜和上皮细胞信号的伴随传输， 参与组织细胞分化和形态发生。使用 结合细胞和分子生物学方法，我们提出， 为了进一步发展这一战略， 用于生物医学研究的非哺乳动物体内模型：1.到 检查正常基底膜形成的过程， 开发和确定基底膜蛋白的确切作用 组织细胞分化和形态发生的结构域。2.审查 葡萄糖水平升高导致异常的 糖尿病患者基底膜增厚 糖尿病。

Basic errors in the structure and function of extracellular matrix (or its specialized form basement membrane) leads to a broad spectrum of problems related to developmental abnormalities and secondary complications in such diseases as diabetes mellitus where basement membrane thickening and dysfunction is observed. Our understanding of the cellula mechanisms underlying normal and abnormal basement membrane formation and the role of matrix components in signal transmission during development has been limited by a lack of appropriate in vitro and in vivo models to experimental approach these problems. In order to better approach these problems, we have developed an in vivo cellular model in which basement membrane formation can be experimentally induced between an epithelial bilayer and its formation related to 1) the process of cytodifferentiation and morphogenesis and 2) disease processes which lead to abnormalities in the structure and function of basement membranes. This model has been developed from the invertebrate, Hydra vulgaris which is structurally reduced to an epithelial bilayer with an intervening basement membrane (mesoglea). Building on previously published studies, we have defined experimental conditions in which 1) pellets generated from cells isolated from adult Hydra form an epithelial bilayer, deposit an intervening basement membrane, and subsequently reform an adult Hydra structure within 72-96 hr through the processes of cytodifferentiation and morphogenesis and 2) as related to the disease diabetes mellitus, development of this cell-aggregation-morphogenesis system in the presence of elevated glucose levels leads to an abnormal thickening of basement membrane within 72-96 hr time frame as opposed to months or years as required in mammalian animal model of diabetes. Utilizing this in vivo epithelial/basement membrane model system we propose to test hypotheses directed at the cellular mechanisms involved in the normal and abnormal development of basement membrane and the concomitant transmission of epithelial signal which are involved in tissue cytodifferentiation and morphogenesis. Using a combination of cellular and molecular biology approaches we propose to focus on the following specific aims in order to further develop this nonmammalian in vivo model for application to biomedical research: 1. To examine the process of normal basement membrane formation during development and determine the precise role of basement membrane protein domains in tissue cytodifferentiation and morphogenesis. 2. To examine cellular mechanisms by which elevated glucose levels lead to an abnormal thickening of basement membrane as observed in individuals with diabetes mellitus.