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Development and differentiation of retinal pigment epithelial cells.

视网膜色素上皮细胞的发育和分化。

基本信息

批准号：
63570006
负责人：
HIROSAWA Kazushige
金额：
$ 1.34万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

项目摘要

Chick retinal pigment epithelial cells were studied by immunohistochemistry with monoclonal antibodies obtained in my laboratory. Results are divided into two parts.1. Development of RPE cells :Photoreceptor cells had no inner and outer segments on the 8th day in ovo. The inner segments have been formed on the 15th day and the outer segments on the 19th day. PRE sheets had short and blunt apical processes on the 8th day, elongated and slender ones on the 15th day, and well-developed and melanin-containing processes on the 19th day. RPE of the 19th day had numerous bundles of actin filaments associated with melanin pigments in the basal portion of the apical processes and in the apical cytoplasm. In rhodamine-phalloidin preparations, intense fluorescence was localized in the RPE apical processes and cytoplasm on the 19th day. We concluded that RPE apical processes develop in accordance with the photoreceptor development.2. Monoclonal bodies specific to the endoplasmic reticulum of RPE :Among the antibodies obtained, two (S5D8 and S5H8) stained the cytoplasm of epithelial cells of the animals examined in finally granular pattern and recognized a 63kD protein in retinal pigment epithelial cells. These antibodies, however, did not react with the rest of the chick eye (cornea, iris, ciliary body, lens and sclera) nor with extraocular tissues (liver, kidney, intestine, brain, testis, adrenal gland, heart, gizzard and skeletal muscle). Immunoelectron microscopy revealed that the antigen was localized in the endoplasmic reticulum of retinal pigment epithelial cells.

用本实验室制备的单克隆抗体对鸡视网膜色素上皮细胞进行了免疫组化研究。研究结果分为两部分.视网膜色素上皮细胞的发育：光感受器细胞在卵内第8天已无内外节。内节在第15天形成，外节在第19天形成。第8天的PRE片具有短而钝的顶突，第15天的PRE片细长，第19天的PRE片发达且含有黑色素。第19天的RPE在顶端突起的基部和顶端细胞质中具有许多与黑色素相关的肌动蛋白丝束。在罗丹明鬼笔环肽制剂中，在第19天，强荧光定位于RPE顶端突起和细胞质中。结论：RPE顶端突起的发育与感光细胞的发育一致. RPE内质网特异性单克隆抗体：在获得的抗体中，两种（S5D8和S5H8）以最终颗粒图案染色检查的动物上皮细胞的细胞质，并识别视网膜色素上皮细胞中的63kD蛋白。然而，这些抗体不与鸡眼的其余部分（角膜、虹膜、睫状体、透镜和巩膜）反应，也不与眼外组织（肝、肾、肠、脑、睾丸、肾上腺、心脏、砂囊和骨骼肌）反应。免疫电镜显示，该抗原定位于视网膜色素上皮细胞的内质网。