Cho E Y P, Choi H L, Chan Franky L
Department of Anatomy, the Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People's Republic of China.
Histochem J. 2002;34(11-12):589-600. doi: 10.1023/a:1026032005521.
The present study sought to characterize the expression and distribution of complex glycoconjugates in the rat retina by lectin histochemistry, using a panel of 21 different lectins with different carbohydrate specificities. Paraffin sections of Carnoy-fixed Sprague-Dawley rat eyes were stained with various biotinylated lectins, followed by the streptavidin-peroxidase and glucose oxidase-diaminobenzidine-nickel staining procedures. The results showed that the retinal pigment epithelium was stained intensely with LCA, Jacalin, WFA, S-WGA, PWA, DSA, UEA-I, LTA and PHA-E, suggesting that this epithelium contained glycoconjugates with alpha-Man, alpha-Glc, alpha-Gal/GalNAc, beta-GalNAc, alpha-Fuc, NeuAc and other oligosaccharide residues. The outer and inner segments of the photoreceptor layer showed different lectin binding affinities. The outer segments reacted with S-WGA and GS-II, whereas the inner segments reacted with UEA-II, UEA-I, LTA and MAA, suggesting that the inner segments contained glycoconjugates rich in alpha-Fuc and NeuAc(alpha2,3)Gal residues. PNA labelled specifically the cones and could be used as a specific marker for these photoreceptors. RCA-I, WFA, S-WGA, DSA, MAA and PHA-E reacted with both the outer and inner plexiform layers. On the other hand, UEA-I and LTA specifically labelled the outer plexiform layer, while PNA labelled the inner plexiform layer. The retinal microglial cells were labelled specifically by GS-I-B4 and SNA. Interestingly, we also observed that WFA bound specifically to Müller cells and could be used as a novel marker for this retinal glial cell. The capillaries and larger vessels in the retina and choriocapillaris reacted intensely with GS-I-B4, RCA-I, S-WGA, PWA, DSA and PHA-E. No significant differences in lectin binding were observed in the microvessels at these two sites. In summary, the present study demonstrated the expression patterns of glycoconjugates in the rat retina and that certain lectins could be used as histochemical markers for specific structural and cellular components of the rat retina.
本研究旨在通过凝集素组织化学法，利用一组21种具有不同碳水化合物特异性的凝集素，对大鼠视网膜中复合糖缀合物的表达和分布进行表征。用Carnoy固定的Sprague-Dawley大鼠眼睛的石蜡切片用各种生物素化凝集素染色，随后进行链霉亲和素-过氧化物酶和葡萄糖氧化酶-二氨基联苯胺-镍染色程序。结果表明，视网膜色素上皮细胞被荆豆凝集素（LCA）、红豆凝集素（Jacalin）、荆豆凝集素（WFA）、大豆凝集素（S-WGA）、豌豆凝集素（PWA）、刀豆凝集素（DSA）、荆豆凝集素（UEA-I）、扁豆凝集素（LTA）和红细胞凝集素（PHA-E）强烈染色，表明该上皮细胞含有具有α-甘露糖、α-葡萄糖、α-半乳糖/ N-乙酰半乳糖胺、β-N-乙酰半乳糖胺、α-岩藻糖、唾液酸和其他寡糖残基的糖缀合物。光感受器层的外段和内段表现出不同的凝集素结合亲和力。外段与大豆凝集素（S-WGA）和麦芽凝集素（GS-II）反应，而内段与荆豆凝集素（UEA-II）、荆豆凝集素（UEA-I）、扁豆凝集素（LTA）和麦胚凝集素（MAA）反应，表明内段含有富含α-岩藻糖和唾液酸（α2,3）半乳糖残基的糖缀合物。花生凝集素（PNA）特异性标记视锥细胞，可作为这些光感受器的特异性标志物。蓖麻凝集素（RCA-I）、荆豆凝集素（WFA）、大豆凝集素（S-WGA）、刀豆凝集素（DSA）、麦胚凝集素（MAA）和红细胞凝集素（PHA-E）与外网状层和内网状层均有反应。另一方面，荆豆凝集素（UEA-I）和扁豆凝集素（LTA）特异性标记外网状层，而花生凝集素（PNA）标记内网状层。视网膜小胶质细胞被麦芽凝集素（GS-I-B4）和神经氨酸酶（SNA）特异性标记。有趣的是，我们还观察到荆豆凝集素（WFA）特异性结合穆勒细胞，可作为这种视网膜神经胶质细胞的新型标志物。视网膜和脉络膜毛细血管中的毛细血管和较大血管与麦芽凝集素（GS-I-B4）、蓖麻凝集素（RCA-I）、大豆凝集素（S-WGA）、豌豆凝集素（PWA）、刀豆凝集素（DSA）和红细胞凝集素（PHA-E）反应强烈。在这两个部位的微血管中未观察到凝集素结合的显著差异。总之，本研究证明了大鼠视网膜中糖缀合物的表达模式，并且某些凝集素可作为大鼠视网膜特定结构和细胞成分的组织化学标志物。
Zotero 插件