Stoward P J, Spicer S S, Miller R L
J Histochem Cytochem. 1980 Sep;28(9):979-90. doi: 10.1177/28.9.7410818.
A peanut lectin-horseradish peroxidase (PL-HRP) conjugate has been applied to histochemical staining of paraffin sections of various mouse organs. The PL-HRP conjugate has selectively reacted with secretory bodies, the Golgi zone, and the apical cell surface in various cell types. Some positive sites, including lingual and tracheal serous glands, Brunner's glands, and the brush border of the proximal straight nephron, contained periodic acid-Schiff (PAS)-positive glycoconjugate with no affinity for basic reagents. The stored secretion in these sites was interpreted as containing neutral glycoprotein with terminal galactose residues which could, in part at least, account for the PAS reactivity. Duodenal goblet cells, which exhibited basophilia attributable to sulfate esters, also bound PL-HRP. As the binding was affected by prior sialidase digestion, the secretory glycoprotein in the duodenal goblet cells was judged to contain oligosaccharides with sulfate esters and terminal galactose uncapped by sialic acid. All sites known from their basophilia to form sialomucin failed to stain with the PL-HRP conjugate, but consistently gained reactivity following sialidase digestion and were inferred, therefore, to possess glycoproteins with oligosaccharide side chains containing subterminal galactose and terminal sialic acid. Lingual mucous glands, known to secrete a mucosubstance with basophilic properties indicative of the presence of sulfate esters but not sialic acid, stained with PL-HRP only after sialidase digestion and, accordingly, were reinterpreted as containing both sulfate esters and terminal galactose-sialic acid dimers. Staining of gastric surface epithelium demonstrated a srongly PAS-reactive neutral glycoprotein, and that of goblet cells in the cecum disclosed PAS-positive sulfated glycoprotein. The latter two sites lacked PL-HRP affinity without or with prior sialidase treatment and apparently possessed neither terminal galactose residues nor galactose-sialic acid dimers. PL-HRP affinity was observed exclusively in the Golgi cisternae of some epithelial cells, thus indicating that galactose occurs transiently as a terminal residue in this site. A few histologic sites, such as pancreatic and gastric zymogen cells and renal tubules, were devoid of both PAS reactivity and basophilia indicative of the presence of complex carbohydrate but stained strongly with the PL-HRP conjugate by means which are not understood. Galactose in the PL-HRP solution blocked or reversed the PL-HRP binding in most of the structures with an affinity for the conjugate, supporting the conclusions that the reagent is specific for galactosyl residues.
花生凝集素 - 辣根过氧化物酶（PL - HRP）复合物已应用于各种小鼠器官石蜡切片的组织化学染色。PL - HRP复合物在各种细胞类型中与分泌小体、高尔基体区域和顶端细胞表面发生了选择性反应。一些阳性部位，包括舌和气管浆液腺、布鲁纳腺以及近端直小管的刷状缘，含有高碘酸 - 希夫（PAS）阳性糖缀合物，对碱性试剂无亲和力。这些部位储存的分泌物被解释为含有带有末端半乳糖残基的中性糖蛋白，这至少可以部分解释其PAS反应性。十二指肠杯状细胞表现出由于硫酸酯导致的嗜碱性，也结合PL - HRP。由于这种结合受到预先唾液酸酶消化的影响，因此判断十二指肠杯状细胞中的分泌糖蛋白含有带有硫酸酯和未被唾液酸封端的末端半乳糖的寡糖。所有已知因其嗜碱性而形成涎黏蛋白的部位均未被PL - HRP复合物染色，但在唾液酸酶消化后始终获得反应性，因此推断其具有带有含亚末端半乳糖和末端唾液酸的寡糖侧链的糖蛋白。已知分泌具有指示硫酸酯存在但无唾液酸的嗜碱性黏液物质的舌黏液腺，仅在唾液酸酶消化后才被PL - HRP染色，因此被重新解释为同时含有硫酸酯和末端半乳糖 - 唾液酸二聚体。胃表面上皮的染色显示出强烈的PAS反应性中性糖蛋白，盲肠杯状细胞的染色揭示了PAS阳性硫酸化糖蛋白。后两个部位在未进行或进行预先唾液酸酶处理的情况下均缺乏PL - HRP亲和力，显然既不具有末端半乳糖残基也不具有半乳糖 - 唾液酸二聚体。仅在一些上皮细胞的高尔基池中观察到PL - HRP亲和力，因此表明半乳糖在该部位作为末端残基短暂出现。一些组织学部位，如胰腺和胃的酶原细胞以及肾小管，既没有PAS反应性也没有指示复合碳水化合物存在的嗜碱性，但却以尚不清楚的方式被PL - HRP复合物强烈染色。PL - HRP溶液中的半乳糖在大多数对该复合物有亲和力的结构中阻断或逆转了PL - HRP的结合，支持了该试剂对半乳糖基残基具有特异性的结论。
