REGULATION OF CELL SHAPE AND PHAGOCYTOSIS IN RPE CELLS

这项工作的长期目标是了解 RPE细胞骨架的构建和发育， 它是如何相互作用形成末端网和顶端突起的， 以及它是如何与细胞膜相连的。 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， 包括鉴定这种细胞中的膜蛋白， 具有极性分布，可以调节细胞间的关系， 细胞-基质相互作用 为了实现这些目标，我们计划使用 抗体，通过免疫荧光鉴定组分， 在发育中的小鸡和大鼠RPE中的免疫印迹。 我们将使用 保持膜-细胞骨架附着的透化细胞 用电子显微镜研究顶端突起的结构 显微镜检查包括冷冻断裂。 我们将完整地碘化 细胞，以确定外表面的哪些成分可以 与细胞骨架相互作用。 我们会研究 细胞的顶端突起形成的时间集中 看看特定的蛋白质是否与细胞骨架结合在一起 在这个时候，并确定他们是否是顶端的组成部分， 流程. 我们将从处理前的阶段共培养RPE 不同年龄胚胎的神经视网膜形成，以确定 如果我们能影响过程的形成。 我们将研究 已知的膜蛋白在发展和培养的RPE。 我们将 开发针对RPE细胞表面的单克隆抗体， 用于测定顶端和基底膜蛋白， 参与细胞粘附的蛋白质， 调节膜蛋白，并确定是否有任何抗体 能够抑制吞噬作用。 我们的研究应该能 在支持和吞噬的过程的结构， 光感受器，正常视力的基本活动。 此外，我们希望进一步了解的因素， 参与维持RPE细胞的极性和对Bruch's的粘附 膜的

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.