Brooks D.E., Komaromy A.M., Garcia-Fernandez M.C., Cutler T.J., Samuelson D.A., Kallberg M.E.
Departments of Large and Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32601, USA.
Vet Ophthalmol. 2000;3(2-3):127-132. doi: 10.1046/j.1463-5224.2000.00127.x.
Purpose To use immunohistochemical techniques to identify and localize the structural macromolecules of the extracellular matrix (ECM) of the normal adult equine lamina cribrosa in order to make comparisons to the extracellular matrix of the lamina cribrosa of horses with glaucoma. METHODS: Normal eyes of five adult horses between 5 and 10 years of age were fixed in 10% neutral buffered formalin and embedded in paraffin. Polyclonal rabbit-derived antibodies against human elastin, laminin, fibrillin-1, and collagen types I, III and IV, and polyclonal goat-derived antibodies against collagen type VI were used as primary antibodies. Transverse and longitudinal histologic sections of the optic nerve head and lamina cribrosa were stained using several dilutions of the primary antibodies, biotinylated link antibody, horseradish peroxidase-labeled streptavidin, and 3,3'-diaminobenzidine as a chromogen. The immunohistochemical staining patterns were qualitatively interpreted. RESULTS: The normal adult horse lamina cribrosa labeled positively for collagen types I, III and VI, laminin, elastin and fibrillin. Collagen type VI staining of the laminar ECM was most intense, followed by labeling for collagen types III and I, respectively. Laminar blood vessels were weakly positive for laminin and slightly positive for type IV collagen. The scleral ECM of the laminar insertion zone had more intense labeling for collagen types I and VI than did the laminar plates. CONCLUSIONS: The extracellular matrix of the laminar plates of the adult equine lamina cribrosa is similar to the dog as it consists of elastic and collagen fibers (with collagen types VI, III and I). Both the normal dog and horse lamina display more intense staining of collagen type VI than is found in the ECM of the normal human lamina cribrosa. The macromolecular structure of the equine lamina cribrosa suggests that it is a very resilient structure that may provide some protection to the optic nerve axons during episodes of elevated intraocular pressure.
目的 运用免疫组织化学技术鉴定并定位正常成年马筛板的细胞外基质（ECM）的结构大分子，以便与患青光眼马匹的筛板细胞外基质进行比较。方法：选取5匹5至10岁成年马的正常眼睛，用10%中性缓冲福尔马林固定，石蜡包埋。使用抗人弹性蛋白、层粘连蛋白、原纤蛋白-1以及I、III和IV型胶原蛋白的兔源多克隆抗体，和抗VI型胶原蛋白的山羊源多克隆抗体作为一抗。视神经乳头和筛板的横向及纵向组织学切片，用几种稀释度的一抗、生物素化连接抗体、辣根过氧化物酶标记的链霉亲和素以及3,3'-二氨基联苯胺作为显色剂进行染色。对免疫组织化学染色模式进行定性解读。结果：正常成年马筛板对I、III和VI型胶原蛋白、层粘连蛋白、弹性蛋白和原纤蛋白呈阳性标记。板层ECM的VI型胶原蛋白染色最强，其次分别是III型和I型胶原蛋白的标记。板层血管对层粘连蛋白呈弱阳性，对IV型胶原蛋白呈弱阳性。板层插入区的巩膜ECM对I型和VI型胶原蛋白的标记比对板层更强烈。结论：成年马筛板的板层细胞外基质与犬相似，由弹性纤维和胶原纤维（VI、III和I型胶原蛋白）组成。正常犬和马的筛板对VI型胶原蛋白的染色均比对正常人筛板的ECM中发现的染色更强。马筛板的大分子结构表明它是一个非常有弹性的结构，在眼压升高期间可能对视神经轴突提供一定保护。
