Defective ingestion of rod outer segments by cultured dystrophic rat pigment epithelial cells.
Defective ingestion of rod outer segments by cultured dystrophic rat pigment epithelial cells.
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发表时间：
1983-07
影响因子：
4.4
通讯作者：
M. Chaitin;M. Hall
中科院分区：
文献类型：
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作者：
M. Chaitin;M. Hall
建立了一种测定大鼠色素上皮细胞（PE）吞噬杆外段（ROS）的新方法。使用ROS抗血清和双重免疫荧光标记程序，可以将附着在这些细胞外表面的ROS与已经摄入的ROS区分开来。我们用这种方法研究了正常大鼠和遗传性视网膜营养不良大鼠（RCS大鼠）分离的PE细胞对ROS的吞噬作用。通过这种方法，我们已经能够证明活性氧在营养不良的PE细胞外表面的附着确实在正常程度上发生。然而，只有少量这些结合的ROS随后被摄入，这表明吞噬作用的摄入阶段是有缺陷的。在持续存在ROS的4小时孵育后，正常大鼠PE细胞摄入了约80%与细胞表面结合的ROS。相反，在这段时间之后，只有不到20%的与营养不良的PE细胞结合的ROS被摄入。这些结果以及脉冲追踪实验的结果表明，正常的PE细胞迅速摄取大部分结合的ROS，而营养不良的PE细胞则没有这种快速摄取。在脉冲追踪实验中，ROS在两小时后被冲洗掉，继续孵育而不进一步添加ROS。然而，随着时间的推移，这两种细胞类型都能够缓慢地摄取额外的结合ROS。
A new procedure for assaying the phagocytosis of rod outer segments (ROS) by cultured rat pigment epithelial (PE) cells has been developed. Using an ROS antiserum and a double immunofluorescent labeling procedure, ROS attached to the external surfaces of these cells can be distinguished from those that have already been ingested. We have used this procedure to study the phagocytosis of ROS by PE cells isolated from normal rats and rats with inherited retinal dystrophy (RCS rats). With this approach we have been able to show that the attachment of ROS to the external surfaces of dystrophic PE cells does take place to a normal extent. However, only a small number of these bound ROS are subsequently ingested, demonstrating that the ingestion phase of phagocytosis is defective. After a 4-hr incubation during which ROS are continuously present, normal rat PE cells ingest about 80% of the ROS that have bound to the cell surfaces. In contrast, after this time period, less than 20% of the ROS bound to the dystrophic PE cells have been ingested. These results, as well as the results of pulse-chase experiments in which ROS are rinsed away after two hours and the incubation continued without further addition of ROS, have demonstrated that normal PE cells rapidly ingest most of the bound ROS, whereas the dystrophic PE cells show no such rapid ingestion. Both cell types, however, are able to slowly ingest additional bound ROS with time.