Fujino Y, Tanishima T
Department of Ophthalmology, University of Tokyo School of Medicine, Japan.
Jpn J Ophthalmol. 1987;31(3):384-92.
The distribution of actin in the rabbit cornea was examined by the indirect immunoperoxidase method using mouse monoclonal anti-actin IgM and goat anti-mouse IgM serum conjugated with peroxidase, which was stained with 4-chloronaphthol. Flat preparation of the cornea was made and the actin distribution was examined in the normal endothelium and also during wound-healing of the endothelium that took place after transcorneal freezing. In the normal cornea, the epithelial, stromal and endothelial cells contained actin in the cytoplasm. Diffuse actin staining in the endothelial cytoplasm was observed in the flat preparation of the normal endothelium. After endothelial damage, the cells adjacent to the damaged area showed a fiber-like staining of actin around the nucleus; this occurred 12 to 24 hours after the freezing. Punctate staining was seen on the 2nd day, and the intensity of the staining was maximum on the 3rd day when the wound was completely covered. On the 3rd day, fibroblast-like cells were encountered and a long fiber-like staining of actin was seen in the cytoplasm, but the staining disappeared on the 4th day. The long fiber-like and punctate staining reappeared on the 14th day. Normal staining pattern was recovered on the 28th day. These findings suggest that actin production was accelerated in the endothelial cells during the wound-healing, and that this productive activity returned to normal levels as healing was completed.
采用间接免疫过氧化物酶法,使用小鼠单克隆抗肌动蛋白IgM和与过氧化物酶结合的山羊抗小鼠IgM血清,以4-氯萘酚染色,检测兔角膜中肌动蛋白的分布情况。制作角膜平铺标本,观察正常内皮细胞以及经角膜冷冻后内皮细胞伤口愈合过程中肌动蛋白的分布。在正常角膜中,上皮细胞层、基质层和内皮细胞的细胞质中均含有肌动蛋白。在正常内皮细胞的平铺标本中,可观察到内皮细胞质中有弥漫性的肌动蛋白染色。内皮损伤后,受损区域附近的细胞在细胞核周围呈现出纤维状的肌动蛋白染色;这种情况在冷冻后12至24小时出现。在第2天可见点状染色,在伤口完全覆盖的第3天染色强度达到最大。在第3天,可见成纤维细胞样细胞,其细胞质中有长纤维状的肌动蛋白染色,但在第4天染色消失。在第14天,长纤维状和点状染色再次出现。在第28天恢复正常染色模式。这些结果表明,在伤口愈合过程中,内皮细胞中的肌动蛋白生成加速,并且随着愈合完成,这种生成活性恢复到正常水平。