Department of Cell Biology and Histology, Faculty of Medicine and Nursing, University of the Basque Country UPV/EHU, Bº Sarriena S/N, 48940, Leioa, Vizcaya, Spain.
Department of Cell Biology and Histology, Regional Campus of International Excellence "Campus Mare Nostrum", IMIB-Arrixaca, University of Murcia, Espinardo, Murcia, Spain.
Anat Sci Int. 2021 Mar;96(2):221-230. doi: 10.1007/s12565-020-00578-4. Epub 2020 Oct 8.
Many studies have been conducted to determine the composition of the glycoconjugates of the mucus-secreting cells of the fundic glands of the stomach. However, the chief cells of these glands have been largely ignored because they secrete mainly zymogens with a lower glycosylation. The aim of this work was to analyze the glycoconjugates of the gastric chief cells by a battery of 17 different lectins, recognizing Fucose, N-acetylgalactosamine, Galactose, N-acetylneuraminic acid, N-acetylglucosamine and Mannose containing oligosaccharides. Histochemical techniques were performed with several lectins and also combined with two pre-treatments; β-elimination, which removes O-linked oligosaccharides, and incubation with Peptide-N-Gycosidase F, which removes N-linked oligosaccharides. In addition, acid hydrolysis was performed before WGA histochemistry, and incubation with glucose oxidase before Con A labeling. Many lectins did not stain the chief cells. In addition, the presence of O-glycans in the apical cell membrane was demonstrated with the lectins AAL, HPA, MPA/MPL, PNA, RCA-I, and WGA. Some of these O-glycans were resistant to short-term β-elimination pre-treatments. Mannose-binding lectins stained the basal cytoplasm of the chief cells. The level of glycosylation of the chief cells was lower than that of the mucous cells. The presence of O-glycans in the apical cell membrane is consistent with the presence of mucins such as MUC1 in the apical membrane of chief cells. Moreover, Mannose-binding lectins revealed N-glycosylation in the basal cytoplasm. The knowledge of gastric chief cell glycoconjugates is relevant because of their potential involvement not only in in physiological but also in pathological processes, such as cancer.
许多研究已经确定了胃底腺黏液分泌细胞糖缀合物的组成。然而,这些腺体的主细胞在很大程度上被忽视了,因为它们主要分泌具有较低糖基化的酶原。本工作的目的是通过 17 种不同的凝集素分析胃主细胞的糖缀合物,这些凝集素识别岩藻糖、N-乙酰半乳糖胺、半乳糖、N-乙酰神经氨酸、N-乙酰葡萄糖胺和甘露糖。采用几种凝集素进行组织化学技术,并结合两种预处理方法;β-消除,去除 O-连接的寡糖,以及用肽-N-糖苷酶 F 孵育,去除 N-连接的寡糖。此外,在 WGA 组织化学之前进行酸水解,并在 Con A 标记之前用葡萄糖氧化酶孵育。许多凝集素没有染色主细胞。此外,用 AAL、HPA、MPA/MPL、PNA、RCA-I 和 WGA 等凝集素证明了顶细胞膜上存在 O-聚糖。这些 O-聚糖中的一些对短期β-消除预处理有抗性。甘露糖结合凝集素染色主细胞的基底细胞质。主细胞的糖基化水平低于黏液细胞。顶细胞膜上存在 O-聚糖与主细胞顶膜上存在粘蛋白(如 MUC1)一致。此外,甘露糖结合凝集素揭示了基底细胞质中的 N-糖基化。胃主细胞糖缀合物的知识是相关的,因为它们不仅在生理过程中,而且在病理过程中,如癌症,都可能涉及到。