Chan F L, Choi H L
Department of Anatomy, Faculty of Medicine, Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.
Histochem Cell Biol. 1995 Nov;104(5):369-81. doi: 10.1007/BF01458131.
The structural organization of integral and associated components of the ciliary zonule is still not fully understood. The present study is to localize and characterize the proteoglycans associated with the ciliary zonule of the rat eye by Cuprolinic blue (CB) staining and immunocytochemistry. After CB staining, the proteoglycans appeared as electron dense elongated rodlets and were localized with the zonular fibers. They were seen lying on the periphery of the zonular fibers or along the length of the individual fibrils. Most of the CB rodlets had a size of 60-170 nm long (average 130 nm) and 25 nm wide. Smaller CB rodlets measuring 25-60 nm long (average 45 nm) and 12 nm wide were sometimes found associated with the individual zonular fibrils. The CB rodlets were removed after chondroitinase ABC or chondroitinase AC treatment, but were resistant to heparitinase, nitrous acid, keratanase or Streptomyces hyaluronidase digestions. The ciliary zonule was also immunostained with three monoclonal antibodies: 2-B-6 specific for chondroitin 4-sulfate, 3-B-3 for chondroitin 6-sulfate and 1-B-5 for unsulfated chondroitin, using indirect immunoperoxidase or immuno-colloidal gold methods. The zonular fibers were immunoperoxidase stained and immunogold labeled by 2-B-6, but were not reactive to 3-B-3 and 1-B-5. The results demonstrate that chondroitin sulfate proteoglycan is associated with the ciliary zonule of the rat eye.
睫状小带的整体及相关组成部分的结构组织仍未被完全理解。本研究旨在通过铜啉蓝(CB)染色和免疫细胞化学方法对大鼠眼睫状小带相关的蛋白聚糖进行定位和表征。CB染色后,蛋白聚糖呈现为电子致密的细长杆状结构,并定位于小带纤维处。它们位于小带纤维的周边或沿着单个纤维的长度分布。大多数CB杆状结构长60 - 170nm(平均130nm),宽25nm。有时会发现与单个小带纤维相关的较小的CB杆状结构,其长25 - 60nm(平均45nm),宽12nm。用软骨素酶ABC或软骨素酶AC处理后,CB杆状结构消失,但对肝素酶、亚硝酸、角质酶或透明质链霉菌酶消化具有抗性。还用三种单克隆抗体对睫状小带进行免疫染色:用间接免疫过氧化物酶或免疫胶体金法,2 - B - 6针对硫酸软骨素4,3 - B - 3针对硫酸软骨素6,1 - B - 5针对未硫酸化的软骨素。小带纤维经2 - B - 6免疫过氧化物酶染色和免疫金标记,但对3 - B - 3和1 - B - 5无反应。结果表明硫酸软骨素蛋白聚糖与大鼠眼的睫状小带相关。