Department of Ophthalmology, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Hirokoji-agaru, Kawaramachi-dori, Kamigyo-ku, Kyoto, 602-0841, Japan.
Department of Frontier Medical Science and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Hirokoji-agaru, Kawaramachi-dori, Kamigyo-ku, Kyoto, 602-0841, Japan.
Sci Rep. 2024 Sep 27;14(1):22391. doi: 10.1038/s41598-024-73145-w.
Age-related macular degeneration (AMD) is associated with the dysfunction and degeneration of retinal pigment epithelium (RPE) cells. Here, we examined how the formation and expansions of cell clusters are regulated by the differentiation of the RPE cells. In this study, ARPE-19 cells were cultivated in standard or differentiation media, i.e., without or with nicotinamide, to evaluate the spreading of cell clusters specified with differentiated cell phenotypes. Mitochondria membrane potential (MMP) and the distribution of the RPE cell clusters was also monitored with or without rotenone, a mitochondrial electron transport chain (ETC) complex I inhibitor. Cultured ARPE-19 cells generated scattered cell clusters composed mostly of smaller size cells expressing the differentiation markers mouse anti-cellular retinaldehyde-binding protein (CRALBP) and Bestrophin only in differentiation medium. After the increase of the number of clusters, the clusters appeared to paracellularly merge, resulting in expansion of the area occupied by the clusters. Of note, the cells within the clusters selectively had high MMP and were in accordance with the expression of RPE differentiation markers. Rotenone repressed the formation of the clusters and decreased intracellular MMP. The above results suggest that clustering of RPE cells with functional mitochondria plays a pivotal role in RPE cell differentiation process and the ETC complex I inhibition greatly influences the composition of RPE cells that are degenerated or differentiation disposed.
年龄相关性黄斑变性（AMD）与视网膜色素上皮（RPE）细胞的功能障碍和变性有关。在这里，我们研究了 RPE 细胞的分化如何调节细胞簇的形成和扩展。在这项研究中，ARPE-19 细胞在标准或分化培养基中培养，即没有或有烟酰胺，以评估具有分化细胞表型的细胞簇的扩散。还监测了线粒体膜电位（MMP）和 RPE 细胞簇的分布，有无鱼藤酮，一种线粒体电子传递链（ETC）复合物 I 抑制剂。培养的 ARPE-19 细胞产生了散在的细胞簇，这些细胞簇主要由较小的表达分化标志物鼠抗细胞视黄醛结合蛋白（CRALBP）和 Bestrophin 的细胞组成，仅在分化培养基中。在细胞簇数量增加后，细胞簇似乎通过细胞旁融合，导致细胞簇所占面积扩大。值得注意的是，簇内的细胞选择性地具有高 MMP，并与 RPE 分化标志物的表达一致。鱼藤酮抑制了细胞簇的形成并降低了细胞内的 MMP。上述结果表明，具有功能线粒体的 RPE 细胞的聚类在 RPE 细胞分化过程中起着关键作用，ETC 复合物 I 的抑制极大地影响了 RPE 细胞的组成，这些细胞退化或分化处理。
