Gómez-Santos Laura, Alonso Edurne, Díaz-Flores Lucio, Madrid Juan F, Sáez Francisco J
Department of Cell Biology and Histology, Training and Research Unit: Reproduction, Development, Aging and Cancer (TRU/UFI 11/44), School of Medicine and Nursing, University of the Basque Country UPV/EHU, Leioa, Vizcaya, Spain (LG-S, EA, FJS).
Department of Anatomy, Pathology, Histology and Radiology, University of La Laguna, San Cristóbal de La Laguna, Tenerife, Spain (LD-F).
J Histochem Cytochem. 2017 May;65(5):261-272. doi: 10.1369/0022155417694871. Epub 2017 Feb 1.
Parietal cells undergo a differentiation process while they move from the isthmus toward the pits and the base region of the gastric gland. The aim of this work was to analyze the rat gastric glands by lectin histochemistry to show the glycans expressed by upper (young) and lower (old) parietal cells. We used lectins recognizing the most frequent sugar moieties in mammals. Each lectin was assayed alone and in combination with several deglycosylation pretreatments: (1) β-elimination, which removes O-linked oligosaccharides; (2) incubation with Peptide-N-glycosidase F, to remove N-linked glycans; (3) acid hydrolysis, which removes terminal sialic acid moieties; (4) methylation-saponification, to remove sulfate groups from sugar residues; and (5) glucose oxidase, a technique carried out with the lectin concanavalin A to convert glucose into gluconic acid. The lectins from Helix pomatia, Dolichos biflorus (DBA), Glycine max (soybean), Maclura pomifera, Arachis hypogaea (peanut), Bandeiraea simplicifolia (lectin I-B), and Datura stramonium showed a different glycan expression in the parietal cells throughout the gastric gland. This difference supports that parietal cells undergo a maturation/degeneration process while the cells descend along the gland. The role of DBA as a marker of parietal cells previously reported should be taken with caution because these cells showed different reactivity for the lectin, ranging from negative to strong labeling.
壁细胞在从胃峡部向胃小凹和胃腺基部区域移动的过程中会经历一个分化过程。这项工作的目的是通过凝集素组织化学分析大鼠胃腺,以显示上部(年轻)和下部(年老)壁细胞表达的聚糖。我们使用了识别哺乳动物中最常见糖部分的凝集素。每种凝集素单独进行检测,并与几种去糖基化预处理方法联合使用:(1)β-消除法,可去除O-连接的寡糖;(2)用肽-N-糖苷酶F孵育,以去除N-连接的聚糖;(3)酸水解,可去除末端唾液酸部分;(4)甲基化-皂化法,从糖残基上去除硫酸基团;(5)葡萄糖氧化酶法,该技术与伴刀豆球蛋白A凝集素一起进行,将葡萄糖转化为葡萄糖酸。来自欧洲滨螺、双花扁豆(DBA)、大豆、桑橙、花生、单叶蔓荆(凝集素I-B)和曼陀罗的凝集素在整个胃腺的壁细胞中显示出不同的聚糖表达。这种差异支持壁细胞在沿腺体下降时会经历成熟/退化过程。先前报道的DBA作为壁细胞标志物的作用应谨慎看待,因为这些细胞对该凝集素显示出不同的反应性,从阴性到强阳性标记不等。