Schraermeyer U, Enzmann V, Kohen L, Addicks K, Wiedemann P, Heimann K
Department of Vitreoretinal Surgery, University of Cologne, Joseph Stelzmann Str. 9, Cologne, 50931, Germany.
Exp Eye Res. 1997 Aug;65(2):277-87. doi: 10.1006/exer.1997.0339.
This study investigates the ability of iris epithelial cells (IPE) to ingest rod outer segments (ROS) and compares the amount of phagocytosis of porcine RPE and IPE cells by the use of a pH sensitive fluorescent dye (carboxy SNAFL) at the light microscopic level. The dye allowed investigation of ingestion separately from binding of rod outer segments. In a second set of experiments, after exposing ferritin-labeled ROS to the cultured cells, phagosomes were also counted in electron microscopic sections. Additionally immunocytochemical staining was performed with IPE and RPE cells. Both cell types stained positive with polyclonal NaK-ATPase antibodies against the alpha 1 subunit from rat brain and kidney. The epithelial nature of the cultured cells was determined by monoclonal anti-human-cytokeratin antibodies. Moreover, the ultrastructure of the cells revealed high amounts of phagosomes smaller than 1 micron in diameter present in both RPE and IPE cells. The iron label of the phagosomes was determined by EELS spectra taken from individual phagosomes. Electron and light microscopic quantification shows that cultured IPE cells have 64% of the phagocytic capacity of the RPE with respect to phagosomes larger than 1 micron in diameter.
本研究调查了虹膜上皮细胞(IPE)摄取视杆细胞外节(ROS)的能力,并通过使用pH敏感荧光染料(羧基SNAFL)在光学显微镜水平比较了猪视网膜色素上皮(RPE)细胞和IPE细胞的吞噬量。该染料能够分别研究摄取情况与视杆细胞外节的结合情况。在第二组实验中,将铁蛋白标记的ROS暴露于培养细胞后,还在电子显微镜切片中对视泡进行了计数。此外,对IPE细胞和RPE细胞进行了免疫细胞化学染色。两种细胞类型都用针对大鼠脑和肾α1亚基的多克隆NaK - ATP酶抗体染色呈阳性。培养细胞的上皮性质通过单克隆抗人细胞角蛋白抗体确定。此外,细胞的超微结构显示RPE细胞和IPE细胞中均存在大量直径小于1微米的视泡。通过从单个视泡获取的电子能量损失谱(EELS)确定视泡的铁标记。电子显微镜和光学显微镜定量显示,就直径大于1微米的视泡而言,培养的IPE细胞的吞噬能力为RPE细胞的64%。