Kato T, Nakayasu K, Kanai A, Nishiyama T, Imamura Y, Hayashi T
Department of Ophthalmology, Juntendo University, Tokyo, Japan.
Ophthalmic Res. 2000 Sep-Oct;32(5):215-21. doi: 10.1159/000055616.
Streptavidin peroxidase immunohistochemistry and immunoelectron microscopy were used to determine the localization of type XII collagen in sections from bovine corneas. Type XII collagen extracted from bovine cornea and skin was assayed by Western blotting. Immunohistochemical experiments showed that type XII collagen was restricted to the corneal stroma; it was not present in corneal epithelium, epithelial basement membrane, Descemet's membrane or endothelium. Type XII collagen was distributed throughout the corneal stroma, and it was prominently localized at the superficial stroma. Immunoelectron-microscopic examination demonstrated that type XII collagen was regularly found along the surface of banded collagen fibrils with a periodic distribution. By Western blot analysis, we observed that extracts from bovine cornea contained both the long and short isoforms of type XII collagen, whereas extracts from bovine skin contained only the short isoform. The homogeneous distribution and/or presence of the long isoform of type XII collagen may be related to the characteristically regular arrangement of collagen fibrils and thereby the transparency of corneal tissue.
采用抗生物素蛋白过氧化物酶免疫组织化学和免疫电子显微镜技术来确定牛角膜切片中Ⅻ型胶原的定位。通过蛋白质免疫印迹法检测从牛角膜和皮肤中提取的Ⅻ型胶原。免疫组织化学实验表明,Ⅻ型胶原局限于角膜基质;在角膜上皮、上皮基底膜、Descemet膜或内皮中不存在。Ⅻ型胶原分布于整个角膜基质,且在浅层基质中显著定位。免疫电子显微镜检查显示,Ⅻ型胶原沿带状胶原纤维表面呈规则分布且具有周期性。通过蛋白质免疫印迹分析,我们观察到牛角膜提取物中同时含有Ⅻ型胶原的长亚型和短亚型,而牛皮肤提取物中仅含有短亚型。Ⅻ型胶原长亚型的均匀分布和/或存在可能与胶原纤维典型的规则排列有关,进而与角膜组织的透明度有关。