ROLE OF SUGARS IN PHAGOCYTOSIS OF ROD OUTER SEGMENTS

糖在杆外节吞噬作用中的作用

• 批准号: 3258110
• 负责人: EDWARD L KEAN
• 金额: $ 17.27万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-05-01 至 1992-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3258110
• 关键词: affinity chromatography; antiidiotype antibody; biotechnology; carbohydrate structure; electron microscopy; embryo /fetus tissue /cell culture; enzyme substrate; liposomes; membrane structure; monoclonal antibody; nonmammalian vertebrate embryology; phagocytosis; radioimmunoassay; retinal pigment epithelium; retinaldehyde; rhodopsin; rod cell; tissue /cell culture

We are investigating the mechanisms involved in the interaction between rhodopsin-containing membranes and the cells of the retinal pigment epithelium (RPE). Little is known of the processes involved in regulating this key phase of the phenomenon of rod renewal. Preliminary evidence has been obtained indicating the presence of receptors for rhodopsin on the RPE surface. The basic experimental design for these studies involves the use of RPE cells of the embryonic chick maintained in cell culture as the biological systems, and rhodopsin-liposomes, rod outer segments (ROS), and disc membranes prepared from the ROS as the target rhodopsin- containing membranes. After incubation under various conditions, the association between the cells and the membranes will be quantitated by means of a radioimmunoassay for rhodopsin. Interference in this process has been observed upon coating the membranes with Fab fragments obtained from polyclonal anti-rhodopsin IgG. Monoclonal antibodies raised against bovine rhodopsin will be characterized in terms of reactivity against specific amino acid sequences of rhodopsin, and Fab fragments prepared from them. The influence of these molecules on the binding may provide information concerning domains of rhodopsin recognized by the RPE. We have observed the binding of rhodopsin and rhodopsin- containing membranes by membranes from bovine RPE. We will explore the nature of this binding, its specificity, and attempt to characterize the components on the RPE that mediate this effect. All of these studies are directed toward investigating whether or not there are receptors for rhodopsin on the RPE. If receptors for rhodopsin are revealed, their isolation will be attempted using rhodopsin affinity chromatoraphy and antiidiotype-antibody technology. We will explore further the possibility that glycosylation of "receptors" on the RPE are involved in the binding of rhodopsin- membranes. The effects of various inhibitors of glycosylation on the process will be examined, such as tunicamycin, swainosonine and castanospermine. Further studies will be carried out dealing with the influence of various carbohydrates on the binding process, using RPE and mouse macrophage cells, with special attention paid to the effect of neoglycoproteins.

我们正在研究这种相互作用的机制 在含有视紫红质的膜和细胞之间 视网膜色素上皮（RPE）。 关于这一点，我们知之甚少。 参与调节这一关键阶段的过程 杆更新现象。 初步证据显示 获得表明视紫红质受体的存在， RPE表面。 这些基本的实验设计 研究涉及使用鸡胚的RPE细胞 维持在细胞培养物中作为生物系统，和 视紫红质-脂质体、视杆外节（ROS）和视盘 由ROS作为目标视紫红质制备的膜- 含有膜。 在各种条件下孵育后， 条件下，细胞和细胞膜之间的联系 将通过放射免疫测定法进行定量， 视紫红质 这一过程中的干扰已被观察到， 用从以下获得的Fab片段包被膜： 多克隆抗视紫红质IgG。 的单克隆抗体 针对牛视紫红质的特性将根据 对视紫红质的特定氨基酸序列的反应性， 以及由它们制备的Fab片段。 这些影响 结合上的分子可以提供关于 由RPE识别的视紫红质结构域。 我们观察到视紫红质和视紫红质的结合- 含有牛RPE膜的膜。 我们将 探索这种结合的性质，它的特异性，并试图 表征RPE上介导这一过程的组分 效果 所有这些研究都是为了调查 视网膜色素上皮上是否有视紫红质受体。 如果 视紫红质受体的揭示，他们的分离将是 尝试使用视紫红质亲和色谱法， 抗独特型抗体技术。 我们将进一步探讨糖基化的可能性， RPE上的“受体”参与视紫红质的结合， 膜。 各种糖基化抑制剂对 将对该工艺进行考察，如衣霉素、苦马豆素 和栗精胺。 将进行进一步的研究，处理的影响， 各种碳水化合物的结合过程中，使用RPE和 小鼠巨噬细胞，特别注意 新糖蛋白的作用。