The glycosylation status of murin postnatal thymus:: a study by histochemistry and lectin blotting
The glycosylation status of murin postnatal thymus:: a study by histochemistry and lectin blotting
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DOI：
10.1007/s10735-008-9180-3
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发表时间：
2008-08-01
影响因子：
3.2
通讯作者：
Sahin, Mesut
中科院分区：
文献类型：
--
作者：
Balcan, Erdal;Guemues, Ayca;Sahin, Mesut
在胸腺发育过程中，胸腺细胞的命运在很大程度上取决于CD4/CD8标志物的变化和T细胞受体蛋白的表达。此外，细胞表面糖基化状态的改变也影响胸腺细胞的成熟。本研究用组织化学和凝集素印迹技术研究了1日龄、9日龄、13日龄和16日龄小鼠胸腺组织糖基化的变化。用pH 5.7的阿尔新蓝(AB)/高碘酸-席夫(PAS)染色，发现胸腺微环境含有羧化和硫酸化的糖胺多聚糖(GAG)。在一些髓质胸腺细胞中可见对唾液粘蛋白特异的AB在pH 2.5时呈强阳性。同样地，研究表明，山毛榉凝集素(MAL)的髓质胸腺细胞，而不是皮质胸腺细胞，含有α(2->；3)连接的唾液酸结构。另一方面，与花生凝集素(PNA)的反应仅见于皮质胸腺细胞，而与花生凝集素(PNA)的反应仅见于皮质胸腺细胞，而与末端甘露糖残基的凝集素(GNA)则见于皮质和髓质。然而，曼陀罗凝集素(DSA)识别半乳糖β(1->；4)N-乙酰氨基葡萄糖，它不仅是细胞特异性的，而且还结合了一些胸腺血管。用凝集素印迹法检测到5条分子量分别为39、54、100、110和212的糖蛋白条带，分别与MAL、PNA、DSA和GNA反应。这些结果表明，在出生后的胸腺细胞选择过程中，细胞表面糖结合物的糖基化模式发生了改变。
During the intrathymic development, the fate of the thymocytes depends largely on variable expression of CD4/CD8 markers and T cell receptor protein expressions. In addition, changes of cell surface glycosylation status also affect the thymocyte maturation. In this study the glycosylation alterations in thymic tissues from 1, 9, 13 and 16 days old mice were evaluated by histochemical and lectin blotting techniques. With alcian blue (AB) at pH 5.7/periodic acid-Schiff (PAS) stainings, it was shown that thymic microenvironments contained carboxlylated and sulfated glycosaminoglycans (GAGs). Strong positivity to AB at pH 2.5, which specific for sialomucins, was seen in some medullary thymocytes. Similarly, it was shown that with Maackia amurensis agglutinin (MAL) medullary thymocytes, but not cortical ones, contained alpha(2 -> 3) linked sialic acid structures. On the other hand, while reaction with peanut agglutinin (PNA), which specific for core disaccharide galactose beta(1 -> 3) N-acetylgalactosamine, was only seen in cortical thymocytes, reaction with Galanthus nivalis agglutinin (GNA), which specific for terminal mannose residues, was seen in both cortex and medulla. However, Datura stramonium agglutinin (DSA), which recognizes galactose beta(1 -> 4) N-acetylglucosamine, was not only cell-specific, but it was bound some thymic vessels. With lectin blotting studies, five glycoprotein bands of molecular weights similar to 39, similar to 54, 100, similar to 110 and similar to 212 were found which reacted with MAL, PNA and DSA as well as GNA. These results suggest that glycosylation patterns of cell surface glycoconjugates are modified during thymocyte selection processes of postnatal days.