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ROLE OF SUGARS IN PHAGOCYTOSIS OF ROD OUTER SEGMENTS

糖在杆外节吞噬作用中的作用

基本信息

批准号：
3258115
负责人：
EDWARD L KEAN
金额：
$ 17.44万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1981
资助国家：
美国
起止时间：
1981-05-01 至 1993-09-29
项目状态：
已结题

项目摘要

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视神经细胞之间视网膜色素上皮(RPE)。人们对此知之甚少监管这一关键阶段所涉及的过程棒材更新现象。初步证据表明表明视紫红质受体的存在 RPE曲面。这些的基本实验设计研究涉及使用胚胎鸡的RPE细胞在细胞培养中作为生物系统保持，以及视紫红质脂质体、视杆外段(ROS)和视盘以ROS为目标视紫红质制备的膜- 含有膜的。在不同的环境下孵化后条件，细胞和膜之间的联系将通过放射免疫分析的方法进行定量视紫红质。在这一过程中观察到的干扰用从以下来源获得的Fab片段涂覆膜多克隆抗视紫质免疫球蛋白。产生的单抗抗牛视紫红质的特征是对视紫红质的特定氨基酸序列的反应性，以及从它们制备的FAB片段。这些因素的影响结合上的分子可以提供关于 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。我们会探索这种绑定的性质及其特殊性，并尝试描述RPE上中介此过程的组件的特征效果。所有这些研究都是针对调查 RPE上是否有视紫红质受体。如果视紫红质受体被发现，它们的分离将是尝试使用视紫红质亲和层析和抗独特型抗体技术。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。不同糖基化抑制剂对血管内皮细胞生长的影响将对该工艺进行检测，如衣霉素、苦马豆碱和蓖麻精粉。将进行进一步的研究，以处理各种碳水化合物在结合过程中，使用RPE和小鼠巨噬细胞，特别注意新糖蛋白的作用。