A dual staining method for identifying mucins of different gastric epithelial mucous cells

Abstract

A dual staining method has been developed to identify two types of mucous secreting cells in the gastric mucosa of human and rat in one and the same tissue section. Sections were stained first using the galactose oxidase-cold thionin Schiff (GOCTS) procedure and then with paradoxical Concanavalin A staining (PCS). Surface mucous cell mucin stained blue with GOCTS, whereas gland mucous cell mucin stained brown with PCS. This method enabled us to differentiate these two types of mucins not only in gastric epithelial cell cytoplasm but also in the extracellular space. Sugar residues detected by GOCTS were explored by employing four species of lectins, which were peanut and Allomyrina dichotoma agglutinins for beta-galactose and Vicia villosa and Wistaria floribunda agglutinins for beta-N-acetylgalactosamine. The effect of oxidation with galactose oxidase was also examined on the affinities of reactive sites for these lectins. The results indicated that, in the human stomach, the sugar residues responsible for this reactivity were most likely beta-N-acetylgalactosamine and beta-galactose in specimens lacking secretion of blood group determinants and beta-N-acetylgalactosamine in those showing the secretion. In the rat stomach, on the other hand, sugar residues responsible for GOCTS were not elucidated by these lectins.

Histochemical similarities of mucins produced by Brunner's glands and pyloric glands: A comparative study

“…In contrast, PHA-L, a bi-and tri-antennary complex-type carbohydrate-specific lectin, reacted only with the mucin granules of some of the mucus-producing cells at the gastroduodenal junction in the rodents, rats, voles, and guinea pigs. The presence of complex-type carbohydrates in mucins is rather unusual; however, this rather unusual histochemical reaction pattern of rat gastric mucin has been reported previously (Ota et al, 1991). Our findings, therefore, appear to emphasize the specialisation of rodent mucins with respect to the presence of complex-type carbohydrates.…”

Helicobacter pylori-Induced Chronic Active Gastritis, Intestinal Metaplasia, and Gastric Ulcer in Mongolian Gerbils

“…The reactivity properties to GOTS/PCS of gerbils' stomach resembled those of the gastric mucosa of other mammals (Figure 2d). 11 Thus, surface mucous cells stained blue with GOTS and gland mucous cells stained brown with PCS ( Figure 2d). The mucosal surface was mostly covered by a thin layer of mucus in which gland mucous cell mucins predominated and surface mucous cell mucins were hardly identified ( Figure 2d).…”

“…This method was used to distinguish two types of gastric mucins, surface mucous cell-mucin and gland mucous cell-mucin. 11 In addition, tissue sections were immunostained for H. pylori by using the indirect immunoperoxidase method, with rabbit anti-H. pylori polyclonal antibody (1:20, DAKO, Glostrup, Denmark), and peroxidaselabeled polyclonal anti-rabbit IgG antibody (1:50, DAKO). Furthermore, BrdU labeling (an indicator of epithelial proliferation) was visualized by using mouse monoclonal antiBrdU antibody (1:50, DAKO) as described previously.…”

Pathophysiological Investigation of the Gastric Surface Mucous Gel Layer of Patients with Helicobacter pylori Infection by Using Immunoassays for Trefoil Factor Family 2 and Gastric Gland Mucous Cell-Type Mucin in Gastric Juice

“…These specimens were cut into 5-mm segments, dehydrated, and embedded in paraffin wax. Tissue sections were stained using hematoxylin and eosin (H&E) for morphological observation and with dual stain consisting of galactose oxidase-cold thionine Schiff reaction (GOTS) for SMC-type mucin, and paradoxical Concanavalin A staining (PCS) for GMC-type mucin [1]. Mouse monoclonal anti-gastric GMC antibody (M-GGMC-1, clone HIK1083, 10-fold dilution) (Kanto Chemical, Tokyo, Japan) and mouse monoclonal anti-human TFF2 antibody (20-fold dilution) (NovoCastra, Newcastle-upon-Tyne, UK) were used to localize GMC-type mucin and TFF2, respectively.…”

“…In the gastric mucosa, 2 types of mucous cells are present, namely, surface mucous cells (SMCs) lining the gastric pits and gland mucous cells (GMCs) in the gastric glands (cardiac gland cells, mucous neck cells, and pyloric gland cells) [1]. Mucins secreted from these mucous cells form the surface mucous gel layer (SMGL) covering the luminal surface of the gastric mucosa [2].…”