MODEL FOR EPITHELIAL/BASEMENT MEMBRANE INTERACTIONS

细胞外基质(或其结构和功能)的基本错误 特殊形式的基底膜)导致了广泛的问题 与发育异常和继发性并发症有关 疾病如糖尿病，基底膜增厚和 观察到功能障碍。我们对细胞机制的理解 基础正常和异常基底膜的形成及其作用 矩阵组件在信号传输过程中得到了发展 由于缺乏合适的体外和体内模型来限制 这些问题都是通过实验方法解决的。为了更好地接近这些 问题，我们已经开发了一个体内细胞模型，在这个模型中 膜的形成可以在实验中诱导上皮细胞之间 双层及其形成与1)细胞分化过程 和形态发生以及2)导致畸形的疾病过程 基底膜的结构和功能。这一模式已经被 由无脊椎动物普通九头蛇进化而来，其结构 还原为具有中间基底膜的双层上皮 (中胚层)。在之前发表的研究的基础上，我们定义了 1)从分离的细胞产生颗粒的实验条件 从成体水母细胞形成上皮双层，沉积一层中间层 基底膜，随后在体内重塑成体的水力结构 72-96小时通过细胞分化和形态发生过程以及 2)与糖尿病有关的疾病，发展为 葡萄糖升高时的细胞-聚集-形态发生系统 在72-96小时内导致基底膜异常增厚 时间框架，而不是哺乳动物所要求的几个月或几年 糖尿病模型。利用这种活体上皮/基底膜 我们建议用来测试针对细胞的假设的模型系统 基底细胞正常和异常发育的机制 膜和伴随的上皮信号的传递，这是 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的方法，我们建议 重点关注以下具体目标，以进一步发展这一目标 应用于生物医学研究的非哺乳动物活体模型：1 观察正常基底膜的形成过程 基底膜蛋白的发展及其精确作用的确定 组织细胞分化和形态发生中的结构域。2.检视 血糖水平升高导致异常的细胞机制 糖尿病患者基底膜增厚的观察 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.