Moon Chan Hee, Cho Heeyoon, Kim Yoon Kyung, Park Tae Kwann
Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea.
Department of Ophthalmology, Hanyang University Guri Hospital, Guri, Korea.
J Korean Med Sci. 2017 Feb;32(2):343-351. doi: 10.3346/jkms.2017.32.2.343.
The present study investigated the temporal pattern and cellular localization of nestin in the adult mouse retina with pharmaceutically induced retinal degeneration using N-methyl-N-nitrosourea (MNU). After a single intraperitoneal injection of MNU in 8-week-old C57BL/6 mice, the animals were sacrificed at 1, 3, 5, 7, and 21 days (n = 6, in each stage). The eyes were examined by means of immunohistochemical tests using nestin, ionized calcium-binding adaptor molecule (Iba-1), CD11b, F4/80, and glial fibrillary acidic protein (GFAP). Western blot analysis and manual cell counting were performed for quantification. Nestin expression was increased after MNU administration. Nestin+/Iba-1+ cells were migrated into outer nuclear layer (ONL) and peaked at day 3 post injection (PI). Nestin+/CD11b+ cells were also mainly identified in ONL at day 3 PI and peaked at day 5. Nestin+/F4/80+ cells were shown in the subretinal space and peaked at day 3 PI. Nestin+/GFAP+ cells were distinctly increased at day 1 PI and peaked at day 5 PI. The up-regulation of nestin expression after MNU administration in adult mouse retinal microglia, and monocyte/macrophage suggests that when retinal degeneration progresses, these cells may revert to a more developmentally immature state. Müller cells also showed reactive gliosis and differentiational changes.
本研究利用N-甲基-N-亚硝基脲(MNU)诱导成年小鼠视网膜发生药物性退变,调查巢蛋白在其中的时间模式和细胞定位。对8周龄C57BL/6小鼠单次腹腔注射MNU后,分别于第1、3、5、7和21天处死动物(每个阶段n = 6)。通过使用巢蛋白、离子钙结合衔接分子(Iba-1)、CD11b、F4/80和胶质纤维酸性蛋白(GFAP)的免疫组织化学检测对眼睛进行检查。进行蛋白质免疫印迹分析和手动细胞计数以进行定量。给予MNU后巢蛋白表达增加。巢蛋白阳性/ Iba-1阳性细胞迁移至外核层(ONL),并在注射后第3天达到峰值。巢蛋白阳性/ CD11b阳性细胞在注射后第3天也主要定位于ONL,并在第5天达到峰值。巢蛋白阳性/ F4/80阳性细胞出现在视网膜下间隙,并在注射后第3天达到峰值。巢蛋白阳性/ GFAP阳性细胞在注射后第1天明显增加,并在注射后第5天达到峰值。成年小鼠视网膜小胶质细胞和单核细胞/巨噬细胞在给予MNU后巢蛋白表达上调,这表明当视网膜退变进展时,这些细胞可能会恢复到发育上更不成熟的状态。穆勒细胞也表现出反应性胶质增生和分化变化。