Neill J M, Thornquist S C, Raymond M C, Thompson J T, Barnstable C J
Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, Connecticut 06510.
Invest Ophthalmol Vis Sci. 1993 Feb;34(2):453-62.
This study sought To learn more about the mechanisms that determine and maintain the differentiated state of the retinal pigment epithelium (RPE).
Monoclonal antibodies were raised against human RPE and used in conjunction with other antibodies. Immunocytochemical and biochemical analyses were performed on tissue sections and cells in culture.
An RPE-specific epitope, RET-PE10, has been detected as a 61 kD cytoplasmic polypeptide in a variety of mammalian, amphibian, and avian species. In the rat, RPE-PE10 was expressed late in eye development, with a faint initial labelling of the RPE in central regions at postnatal day 9 (PN9) that increased to adult levels and extent of staining by PN14. RET-PE10 expression initially was present in overnight cultures of dissociated rat RPE cells but was lost rapidly from these cultures during the first week. Comparison of the staining patterns of RET-PE10 with those of various cytoskeletal elements suggests that RET-PE10 may be associated with part of the intermediate filament network. Culture of whole eyecups also resulted in a loss of RET-PE10 expression. RET-PE10 expression was normal in eyes of adult rd/rd mutant mice.
RET-PE10 is a late-appearing marker of RPE differentiation. The results also suggest that the maintained expression of RET-PE10 depends upon extrinsic factors but that these do not include maintained contact with Bruch's membrane or light-induced retinal activity.
本研究旨在进一步了解决定和维持视网膜色素上皮(RPE)分化状态的机制。
制备针对人RPE的单克隆抗体,并与其他抗体联合使用。对组织切片和培养细胞进行免疫细胞化学和生化分析。
在多种哺乳动物、两栖动物和鸟类中,已检测到一种RPE特异性表位RET-PE10,它是一种61kD的细胞质多肽。在大鼠中,RPE-PE10在眼睛发育后期表达,出生后第9天(PN9)中央区域的RPE最初有微弱标记,到PN14时增加到成年水平并达到染色范围。RET-PE10最初存在于解离的大鼠RPE细胞过夜培养物中,但在第一周内从这些培养物中迅速消失。将RET-PE10的染色模式与各种细胞骨架成分的染色模式进行比较表明,RET-PE10可能与中间丝网络的一部分相关。整个眼杯的培养也导致RET-PE10表达丧失。成年rd/rd突变小鼠眼睛中RET-PE10表达正常。
RET-PE10是RPE分化的晚期出现标记。结果还表明,RET-PE10的持续表达取决于外在因素,但这些因素不包括与布鲁赫膜的持续接触或光诱导的视网膜活动。