Lewis G P, Matsumoto B, Fisher S K
Neuroscience Research Institute, University of California, Santa Barbara 93106, USA.
Invest Ophthalmol Vis Sci. 1995 Nov;36(12):2404-16.
The goal of this study was to determine the changes in the organization of the retinal cytoskeleton after experimental retinal detachment.
Cat retinas were detached from the retinal pigment epithelium and then processed for Western blot analysis and fluorescence microscopy. Proteins examined included glial fibrillary acidic protein (GFAP), vimentin, tubulin, and actin. Sections were viewed using a laser scanning confocal microscope.
GFAP and vimentin: At 1 day after detachment, there was an aggregation of intermediate filaments in the endfoot of Müller cells. At 3 days, intermediate filament containing Müller cell processes could be detected within the subretinal space, and, at 28 days, these processes formed large glial scars in the subretinal space. beta-tubulin: At 3 days after detachment, an increase in immunolabeling could be detected within the Müller cell endfoot and in Müller cell processes within the subretinal space. Actin: At 3 days after detachment, rhodamine-phalloidin staining decreased in the inner segments, the photoreceptor synaptic terminals, and the outer limiting membrane.
The decrease in labeling of the photoreceptor inner segment and synaptic terminal cytoskeleton may be a key indicator of early changes in photoreceptors after detachment. The increase in cytoskeletal proteins GFAP, vimentin, and tubulin within the retinal Müller cells after detachment may help to stabilize this cell type as it hypertrophies during glial scar formation. Inhibition of this response may aid in the treatment of diseases in which Müller cell hypertrophy plays a role.
本研究的目的是确定实验性视网膜脱离后视网膜细胞骨架组织的变化。
将猫的视网膜与视网膜色素上皮分离,然后进行蛋白质印迹分析和荧光显微镜检查。检测的蛋白质包括胶质纤维酸性蛋白(GFAP)、波形蛋白、微管蛋白和肌动蛋白。使用激光扫描共聚焦显微镜观察切片。
GFAP和波形蛋白:脱离后1天,Müller细胞终足中的中间丝聚集。3天时,视网膜下间隙内可检测到含有Müller细胞突起的中间丝,28天时,这些突起在视网膜下间隙形成大的胶质瘢痕。β-微管蛋白:脱离后3天,在Müller细胞终足和视网膜下间隙内的Müller细胞突起中可检测到免疫标记增加。肌动蛋白:脱离后3天,视杆细胞内节、光感受器突触终末和外界膜中的罗丹明-鬼笔环肽染色减少。
光感受器内节和突触终末细胞骨架标记的减少可能是脱离后光感受器早期变化的关键指标。脱离后视网膜Müller细胞内细胞骨架蛋白GFAP、波形蛋白和微管蛋白的增加可能有助于在胶质瘢痕形成过程中该细胞类型肥大时使其稳定。抑制这种反应可能有助于治疗Müller细胞肥大起作用的疾病。