Stein Eye Institute, UCLA David Geffen School of Medicine, Doris Stein Building, 200 Stein Plaza, 90095, Los Angeles, CA, USA,
Adv Exp Med Biol. 2014;801:85-90. doi: 10.1007/978-1-4614-3209-8_11.
The ingestion and degradation of photoreceptor disk membranes is a critical and major role for the retinal pigment epithelium (RPE). To help elucidate the cellular events involved in this role, functional in vivo and in vitro assays need to be developed further. Here we propose a method to help monitor phagosome maturation, using antibodies against different epitopes of opsin. We show that antibodies specific for the C-terminus of opsin label only immature phagosomes located in the apical region of the RPE. In contrast, antibodies recognizing the N-terminus also label more mature phagosomes, located more basally. The combined use of antibodies against different opsin epitopes thus provides a valuable tool in the study of phagosome maturation in the RPE.
光感受器盘膜的摄取和降解是视网膜色素上皮 (RPE) 的关键和主要功能。为了帮助阐明这一作用涉及的细胞事件，需要进一步开发功能的体内和体外测定法。在这里，我们提出了一种使用针对视蛋白不同表位的抗体来帮助监测吞噬体成熟的方法。我们发现，针对视蛋白 C 末端的抗体仅标记位于 RPE 顶区的未成熟吞噬体。相比之下，识别 N 末端的抗体也标记更多位于基底侧的更成熟的吞噬体。因此，使用针对不同视蛋白表位的抗体的联合使用为研究 RPE 中的吞噬体成熟提供了有价值的工具。
