Development and differentiation of retinal pigment epithelial cells.

用本实验室获得的单抗对鸡视网膜色素上皮细胞进行免疫组织化学研究。研究结果分为两部分。视网膜色素上皮细胞发育情况：卵子第8天，感光细胞内、外无节段。内节在第15天形成，外节在第19天形成。第8天根尖突起短而钝，第15天尖突细长，第19天突起发达且含有黑色素。第19天的RPE在顶端突起的底部和顶端细胞质中有大量与黑色素相关的肌动蛋白细丝。在罗丹明-鬼笔花素制剂中，第19天在RPE根尖突起和细胞质中有强烈的荧光。结论：1.视网膜色素上皮尖突的发育与感光细胞的发育相一致。RPE内质网特异的单抗：在获得的抗体中，两种抗体(S5D8和S5H8)染色动物的上皮细胞胞浆，最终呈颗粒状，在视网膜色素上皮细胞中识别出一个63kD的蛋白。然而，这些抗体不与鸡眼的其余部分(角膜、虹膜、睫状体、晶状体和巩膜)或眼外组织(肝、肾、肠、脑、睾丸、肾上腺、心脏、肌腱和骨骼肌)发生反应。免疫电子显微镜显示该抗原定位于视网膜色素上皮细胞的内质网。

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.