Tawara A, Varner H H, Hollyfield J G
Cullen Eye Institute, Baylor College of Medicine, Houston, Tx 77030.
Exp Eye Res. 1988 May;46(5):689-704. doi: 10.1016/s0014-4835(88)80056-3.
Cuprolinic Blue, when applied at a critical electrolyte concentration, can be utilized for assessing the localization and structural characteristics of proteoglycans with electron microscopy. We have used this cytochemical procedure to evaluate the distribution of proteoglycan in the interphotoreceptor matrix of the mouse retina. Cuprolinic Blue-positive filaments of two distinct morphological types were present surrounding both rod and cone photoreceptors. Large filaments, 115-135 nm long and 15-25 nm in diameter, were distributed in the interphotoreceptor matrix around the outer segment and outer portion of the inner segment. These filaments appeared linked to each other to form a complex meshwork. Smaller filaments, 60-70 nm long and 5-10 nm in diameter, were principally observed around the photoreceptor inner segments. Incubation of retinas with chondroitinase AC and chondroitinase ABC eliminated Cuprolinic Blue staining of both large and small filaments, whereas hyaluronidase treatment reduced the size of the filaments but did not eliminate their staining. When retinas were washed extensively prior to fixation and staining, Cuprolinic Blue-positive filaments remained associated with the photoreceptor cell surface. These results suggest that the interphotoreceptor matrix of the mouse retina contains at least two structural types of proteoglycan, of the chondroitin sulfate-type, which are differentially distributed in this compartment. One of the proteoglycans forms a complex meshwork which surrounds the photoreceptors. Both are insoluble and appear to be firmly attached to the photoreceptor plasma membrane.
铜叶绿酸蓝在临界电解质浓度下应用时,可用于通过电子显微镜评估蛋白聚糖的定位和结构特征。我们已使用这种细胞化学方法来评估蛋白聚糖在小鼠视网膜光感受器间基质中的分布。在视杆和视锥光感受器周围均存在两种不同形态类型的铜叶绿酸蓝阳性细丝。较大的细丝长115 - 135纳米,直径15 - 25纳米,分布在外段和内段外部周围的光感受器间基质中。这些细丝似乎相互连接形成一个复杂的网络。较小的细丝长60 - 70纳米,直径5 - 10纳米,主要在光感受器内段周围观察到。用软骨素酶AC和软骨素酶ABC孵育视网膜可消除大小细丝的铜叶绿酸蓝染色,而用透明质酸酶处理可减小细丝大小但不会消除其染色。当视网膜在固定和染色前进行广泛冲洗时,铜叶绿酸蓝阳性细丝仍与光感受器细胞表面相关联。这些结果表明,小鼠视网膜的光感受器间基质含有至少两种硫酸软骨素类型的结构不同的蛋白聚糖,它们在该区域中分布不同。其中一种蛋白聚糖形成围绕光感受器的复杂网络。两者均不溶,且似乎牢固地附着于光感受器质膜。