Okamoto C T, Forte J G
Department of Physiology-Anatomy, University of California, Berkeley.
Gastroenterology. 1988 Aug;95(2):334-42. doi: 10.1016/0016-5085(88)90488-x.
The distribution of lectin-binding sites in oxyntic and chief cells of isolated rabbit gastric glands was determined with seven fluoresceinated lectins, to ascertain which lectins might best be used in the biochemical characterization of cell membranes and glycoproteins of these two cell types. Oxyntic cell canaliculi were labeled by wheat germ, Helix pomatia, and peanut lectins, suggesting a predominance of N-acetylhexosamines. Tubulovesicles were heavily stained by wheat germ, Helix pomatia, and Ricinus communis I lectins, indicative of N-acetylhexosamine- and galactose-containing glycoconjugates. Diffuse oxyntic cell cytoplasmic staining was observed with the mannose-binding lectin concanavalin A. This lectin, along with wheat germ, soybean, Helix pomatia, and Ricinus communis I lectins, bound to oxyntic cell basolateral membranes, indicating mannose, N-acetylhexosamine, and galactose residues. Chief cell apical membranes were labeled with peanut, Ricinus communis I, Helix pomatia, and Ulex europaeus lectins, suggesting a predominance of N-acetylhexosamine, galactose, and fucose residues. None of the lectins demonstrated any significant affinity for chief cell cytoplasm or basolateral membrane. Ulex europaeus agglutinin-binding sites were additionally concentrated in lateral intercellular spaces. The results of this study indicate that the range of utility of isolated rabbit gastric glands can be expanded to include histochemical work. In addition, the data suggest the applicability of lectin affinity chromatography in the isolation and characterization of oxyntic and chief cell membranes and glycoproteins.
用七种荧光素标记的凝集素测定了分离的兔胃腺壁细胞和主细胞中凝集素结合位点的分布，以确定哪些凝集素最适合用于这两种细胞类型的细胞膜和糖蛋白的生化特性研究。壁细胞的微管被麦芽、蜗牛和花生凝集素标记，表明N-乙酰己糖胺占优势。微管泡被麦芽、蜗牛和蓖麻凝集素I重度染色，表明存在含N-乙酰己糖胺和半乳糖的糖缀合物。用结合甘露糖的凝集素伴刀豆球蛋白A观察到壁细胞胞质弥漫性染色。这种凝集素与麦芽、大豆、蜗牛和蓖麻凝集素I一起，与壁细胞基底外侧膜结合，表明存在甘露糖、N-乙酰己糖胺和半乳糖残基。主细胞顶端膜被花生、蓖麻凝集素I、蜗牛和荆豆凝集素标记，表明N-乙酰己糖胺、半乳糖和岩藻糖残基占优势。没有一种凝集素对主细胞胞质或基底外侧膜表现出任何显著亲和力。荆豆凝集素结合位点还集中在细胞间侧隙。这项研究的结果表明，分离的兔胃腺的应用范围可以扩大到包括组织化学工作。此外，数据表明凝集素亲和色谱法在壁细胞和主细胞膜及糖蛋白的分离和特性鉴定中的适用性。
