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软骨基质中糖胺聚糖的组织化学。半薄环氧树脂切片中的阿尔辛蓝染色和凝集素结合亲和力。

Histochemistry of glycosaminoglycans in cartilage ground substance. Alcian-blue staining and lectin-binding affinities in semithin Epon sections.

作者信息

Mallinger R, Geleff S, Böck P

出版信息

Histochemistry. 1986;85(2):121-7. doi: 10.1007/BF00491758.

Abstract

The critical-electrolyte-concentration staining method using Alcian blue (AB) was applied to etched semithin Epon-embedded sections. The distribution of various glycosaminoglycans (GAGs) was studied in hyaline, elastic, cellular and fibrous cartilage obtained from humans and rodents. The staining patterns in semithin sections were found to correspond to those obtained using paraffin-embedded material. Lectin histochemistry was performed on consecutive sections. The following peroxidase-labelled lectins were used: Ricinus communis A I, Arachis hypogaea, Ulex europaeus A I, Triticum vulgaris, Helix pomatia, Limax flavus, and concanavalin A. The lectin-binding capacity of cartilaginous ground substance was found to be low, as was expected on account of the few free sugar residues of GAGs. Chondroitin sulphate, the most widely distributed GAG, did not exhibit lectin staining. The lectin-binding sites (positive staining for all lectins tested except H. pomatia) observed corresponded to areas positive for keratan sulphate, as shown by AB staining in preceding or following sections. The pronounced lectin binding seen in cellular structures and the inner territorial matrix regions is considered to be due to higher concentrations of oligosaccharides involved in the metabolism of GAGs.

摘要

将使用阿尔新蓝(AB)的临界电解质浓度染色方法应用于蚀刻后的半薄环氧树脂包埋切片。研究了从人类和啮齿动物获取的透明软骨、弹性软骨、细胞软骨和纤维软骨中各种糖胺聚糖(GAGs)的分布情况。发现半薄切片中的染色模式与使用石蜡包埋材料所获得的模式一致。对连续切片进行凝集素组织化学分析。使用了以下过氧化物酶标记的凝集素:蓖麻凝集素AI、花生凝集素、荆豆凝集素AI、小麦凝集素、蜗牛凝集素、蛞蝓凝集素和伴刀豆球蛋白A。正如根据GAGs中游离糖残基较少所预期的那样,发现软骨基质的凝集素结合能力较低。分布最广泛的GAG——硫酸软骨素未表现出凝集素染色。观察到的凝集素结合位点(除蜗牛凝集素外,所有测试凝集素均呈阳性染色)与硫酸角质素阳性区域相对应,如先前或后续切片中的AB染色所示。在细胞结构和内部区域基质中观察到的明显凝集素结合被认为是由于参与GAGs代谢的寡糖浓度较高所致。

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