Weinreb R N, Ryder M I
Department of Ophthalmology, University of California, San Diego, La Jolla 92093.
Invest Ophthalmol Vis Sci. 1990 Sep;31(9):1839-47.
The authors compared cytoskeletal elements of the in situ human trabecular-meshwork cell with in situ human corneal cells using indirect immunofluorescence staining for tubulin and intermediate filaments (vimentin, cytokeratin, and desmin) and NBD-phallacidin staining for f-actin using both fixed frozen and unfixed frozen sections from postmortem eyes. Both f-actin and tubulin were found throughout the cell body of trabecular-meshwork cells, keratocytes, corneal endothelium, and corneal epithelium. The f-actin staining pattern was concentrated at the cell periphery of these four cell types. Vimentin stain was intensely localized in focal areas of the trabecular-meshwork cell, keratocytes, and throughout the corneal endothelium. A general anticytokeratin antibody was intensely localized in corneal epithelium and endothelium. However, PKK-1 anticytokeratin antibody was seen only in superficial layers of corneal epithelium and not in corneal endothelium. The 4.62 anticytokeratin antibody was not observed in either corneal epithelium or endothelium. None of these three cytokeratin antibodies were seen in trabecular-meshwork cells or keratocytes. Desmin stain was not noted in any of these cell types. In general, cytoskeletal staining of unfixed frozen sections showed a similar staining pattern for f-actin and tubulin but a more uniform and intense staining pattern for vimentin and cytokeratin compared with fixed frozen material. The authors conclude that these cytoskeletal stains can differentiate human trabecular-meshwork cells from cells of the cornea in situ.
作者使用针对微管蛋白和中间丝（波形蛋白、细胞角蛋白和结蛋白）的间接免疫荧光染色以及针对丝状肌动蛋白的NBD-鬼笔环肽染色，对死后眼睛的固定冷冻切片和未固定冷冻切片进行检测，比较了原位人类小梁网细胞和原位人类角膜细胞的细胞骨架成分。在小梁网细胞、角膜细胞、角膜内皮细胞和角膜上皮细胞的整个细胞体内均发现了丝状肌动蛋白和微管蛋白。这四种细胞类型的丝状肌动蛋白染色模式集中在细胞周边。波形蛋白染色强烈定位于小梁网细胞、角膜细胞的局部区域以及整个角膜内皮。一种通用的抗细胞角蛋白抗体强烈定位于角膜上皮和内皮。然而，PKK-1抗细胞角蛋白抗体仅在角膜上皮的表层可见，而在角膜内皮中未见。4.62抗细胞角蛋白抗体在角膜上皮或内皮中均未观察到。这三种细胞角蛋白抗体在小梁网细胞或角膜细胞中均未见到。在这些细胞类型中均未发现结蛋白染色。总体而言，与固定冷冻材料相比，未固定冷冻切片的细胞骨架染色显示丝状肌动蛋白和微管蛋白的染色模式相似，但波形蛋白和细胞角蛋白的染色模式更均匀且强烈。作者得出结论，这些细胞骨架染色可在原位将人类小梁网细胞与角膜细胞区分开来。
