INFLUENCE OF SULFATE GROUPS IN THE BINDING OF PEANUT AGGLUTININ - HISTOCHEMICAL-DEMONSTRATION WITH LIGHT-MICROSCOPY AND ELECTRON-MICROSCOPY
INFLUENCE OF SULFATE GROUPS IN THE BINDING OF PEANUT AGGLUTININ - HISTOCHEMICAL-DEMONSTRATION WITH LIGHT-MICROSCOPY AND ELECTRON-MICROSCOPY
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DOI：
10.1007/bf01046790
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发表时间：
1992-04-01
期刊：
影响因子：
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通讯作者：
CASTELLS, MT
中科院分区：
文献类型：
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作者：
MARTINEZMENARGUEZ, JA;BALLESTA, J;CASTELLS, MT
本文用组织化学技术在光镜和电镜水平上研究了粘液糖蛋白硫酸化对花生凝集素（PNA）与组织切片结合的影响。采用连续甲基化-皂化程序对组织样品进行纯化。过氧化物酶和胶体金标记的PNA进行了比较，在控制和rhizophated大鼠肠粘膜样品。高铁二胺（HID）技术被用作控制的有效性的澄清技术，和阿尔新蓝，pH 2.5（AB 2.5），PAS和磷钨酸-HCl（酸-PTA）技术作为控制的寡糖链的完整性，分别。一般而言，与对照切片相比，在去磷酸化样品中观察到PNA反应性显著增加。这些发现表明半乳糖的硫酸化抑制PNA与组织切片中的碳水化合物部分的结合。在大肠的杯状细胞中用HID、PNA和双磷酸肽-PNA序列获得的染色模式表明，随着细胞成熟并从下隐窝区域向腔表面移动，储存在这些细胞中的分泌产物发生了改变。在小肠中未检测到这些修饰。PNA结合位点的超微结构检测表明，半乳糖残基被纳入O-连接的糖蛋白在高尔基体的内侧池的寡糖链。然而，硫酸化发生在高尔基体复合体和高尔基体网络的反侧。总之，纯化程序对于揭示PNA结合位点是有用的。
The influence of sulphation of mucus glycoproteins in the binding of peanut agglutinin (PNA) to tissue sections has been investigated by means of histochemical techniques at the light- and electron-microscopic level. A sequential methylation-saponification procedure was applied for the desulphation of tissue samples. Labelling by peroxidase- and colloidal gold-conjugated PNA was compared in control and desulphated samples of rat intestinal mucosa. The high-iron-diamine (HID) technique was used as a control for the effectiveness of the desulphation technique, and the Alcian Blue, pH 2.5 (AB 2.5), PAS and phosphotungstic acid-HCl (acid-PTA) techniques served as controls for the integrity of the oligosaccharide chains, respectively. In general, a marked increase of PNA reactivity was observed in desulphated samples when compared with control sections. These findings indicate that sulphation of galactose inhibits the binding of PNA to carbohydrate moieties in tissue sections. Staining patterns obtained with HID, PNA and the desulphation-PNA sequence in the goblet cells of the large intestine suggest a modification of the secretory product stored in these cells as the cell matures and moves from the lower crypt region toward the luminal surface. These modifications were not detected in the small intestine. Ultrastructural detection of PNA-binding sites suggests that galactose residues are incorporated into the oligosaccharide chains of O-linked glycoproteins at the medial cisternae of the Golgi apparatus. However, sulphation occurs at the trans side of the Golgi complex and the trans Golgi network. In conclusion, desulphation procedures are useful for revealing PNA-binding sites.