Zhang X S, Proctor G B, Garrett J R, Schulte B A, Shori D K
Secretory and Soft Tissue Research Unit, Rayne Institute, King's College School of Medicine and Dentistry, London, United Kingdom.
J Histochem Cytochem. 1994 Sep;42(9):1261-9. doi: 10.1177/42.9.8064133.
We used a panel of nine lectins to detect the glycosylation patterns of rat submandibular glycoproteins. Binding of lectins was assessed on tissue sections and on Western blots of electrophoretically separated glycoproteins from glandular extracts or sympathetic saliva. Histochemical staining of tissue sections showed that two lectins (DBA and SBA) with affinity for terminal GalNAc residues were localized specifically to acinar cells. In contrast, LTA and UEA-I (alpha Fuc-directed) and LFA (NeuAc-directed) bound exclusively to granular tubule cells. PNA and MPA (beta Gal-directed), LCA (alpha Man- and alpha Glc-directed), WGA (beta GlcNAc- and NeuAc-directed), and sWGA (beta GlcNAc-directed) bound to both acinar and granular tubule cells. On electroblot preparations, LFA, PNA, WGA, DBA, and SBA reacted with high molecular weight acinar mucin components both in glandular extracts and in saliva. LTA, UEA-I, LFA, PNA, MPA, LCA, WGA, sWGA, and DBA bound to lower molecular weight bands on blots known to contain granular tubular proteinases. Lectin binding to acini and granular tubules was reduced in sections and in most bands from glandular extracts after sympathetic nerve stimulation. Our results show that (a) secretory glycoproteins from rat submandibular acinar cells are non-fucosylated and contain abundant alpha GalNAc and NeuAc and a small proportion of beta Gal in their oligosaccharide side chains, and (b) alpha Fuc, NeuAc, beta Gal, and alpha GalNAc form the major carbohydrate moieties of the secretory glycoproteins from granular tubules. The results confirm the considerable potential of lectin probes for studying glycoproteins in secretions and in their cells of origin.
我们使用一组九种凝集素来检测大鼠下颌下腺糖蛋白的糖基化模式。在组织切片以及来自腺体提取物或交感神经唾液的经电泳分离的糖蛋白的Western印迹上评估凝集素的结合情况。组织切片的组织化学染色显示，对末端GalNAc残基具有亲和力的两种凝集素（DBA和SBA）特异性定位于腺泡细胞。相比之下，LTA和UEA-I（α-岩藻糖导向）以及LFA（NeuAc导向）仅与颗粒小管细胞结合。PNA和MPA（β-半乳糖导向）、LCA（α-甘露糖和α-葡萄糖导向）、WGA（β-葡萄糖胺和NeuAc导向）以及sWGA（β-葡萄糖胺导向）与腺泡细胞和颗粒小管细胞均结合。在电印迹制剂上，LFA、PNA、WGA、DBA和SBA在腺体提取物和唾液中均与高分子量腺泡粘蛋白成分发生反应。LTA、UEA-I、LFA、PNA、MPA、LCA、WGA、sWGA和DBA与印迹上已知含有颗粒小管蛋白酶的较低分子量条带结合。交感神经刺激后，组织切片和腺体提取物的大多数条带中凝集素与腺泡和颗粒小管的结合减少。我们的结果表明：（a）大鼠下颌下腺腺泡细胞分泌的糖蛋白在其寡糖侧链中是非岩藻糖基化的，含有丰富的α-GalNAc和NeuAc以及一小部分β-半乳糖；（b）α-岩藻糖、NeuAc、β-半乳糖和α-GalNAc构成颗粒小管分泌糖蛋白的主要碳水化合物部分。这些结果证实了凝集素探针在研究分泌物及其起源细胞中的糖蛋白方面具有相当大的潜力。
