REGULATION OF CELL SHAPE AND PHAGOCYTOSIS IN RPE CELLS

这项工作的长期目标是理解 RPE细胞细胞骨架的构建和发育， 它如何相互作用形成终末网络和心尖突起， 以及它是如何连接到膜上的。此外，我们的目标是获得 关于RPE细胞的极性如何发展的信息， 包括鉴定该细胞中的膜蛋白 具有极分布，它可以调节细胞-细胞和 细胞-底物相互作用。为了实现这些目标，我们计划使用 通过免疫荧光识别成分的抗体和 鸡和大鼠视网膜色素上皮发育过程中的免疫印迹。我们将使用 保持膜-细胞骨架连接的通透性细胞 用电子方法研究顶突的结构 包括冷冻断裂在内的显微镜检查。我们会原封不动地加碘 用于确定外表面的哪些成分可以 与细胞骨架相互作用。我们将研究发展 关注顶突形成时间的细胞 以了解特定的蛋白质是否与细胞骨架相关 并确定它们是否是根尖的组成部分 流程。我们将从加工前的各个阶段对RPE进行共培养 不同胎龄胚胎神经视网膜的形成 如果我们能影响过程的形成。我们将研究极性 已知的膜蛋白在RPE的发育和培养中的作用。我们会 开发针对RPE细胞表面的单抗，这将是 用于测定顶膜和基底膜蛋白，鉴定 参与细胞黏附的蛋白质，发育特性 调节的膜蛋白，并确定是否有抗体 能够抑制吞噬作用的。我们的研究应该会让我们明白 关于支持和吞噬细胞的过程的结构 光感受器，是正常视力的基本活动。 此外，我们希望对这些因素有进一步的了解 参与维持RPE细胞的极性和与Bruchs的粘附性 薄膜。

The long term objectives of this work are to understand the construction and development of the cytoskeleton of the RPE cell, how it interacts to form the terminal web and the apical processes, and how it is linked to the membrane. In addition we aim to gain information about how the polarity of the RPE cell develops, including identification of membrane proteins in this cell which have a polar distribution and which may regulate cell-cell and cell-substratum interactions. Towards these goals we plan to use antibodies to identify components by immunofluorescence and immunoblotting in developing chick and rat RPE. We will use permeabilized cells which retain membrane-cytoskeletal attachments for studies of the structure of apical processes by electron microscopy including freeze-fracture. We will iodinate intact cells to determine which components of the external surface may interact with the cytoskeleton. We will study the development of the cells with a focus on the time of formation of apical processes to see if specific proteins become associated with the cytoskeleton at this time, and to determine if they are components of the apical processes. We will coculture the RPE from stages before process formation with neural retina of different aged embryos to determine if we can influence process formation. We will study the polarity of known membrane proteins in developing and cultured RPE. We will develop monoclonal antibodies to RPE cell surfaces which will be used to determine apical and basal membrane proteins, identify proteins involved in cell adhesion, characterize developmentally regulated membrane proteins and determine if any antibodies are capable of inhibiting phagocytosis. Our studies should shed light on the structure of the processes which support and phagocytose the photoreceptors, essential activities for normal vision. Additionally we hope to gain further understanding of the factors involved in maintaining RPE cell polarity and adhesion to Bruch's membrane.