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PubMed2026

Colitogenic Environment Suppresses Notch/Mechanistic Target of Rapamycin-Mediated Epithelial Proliferation to Promote Goblet Cell Differentiation in the Colon.

Epithelial barrier dysfunction is a hallmark of inflammatory bowel diseases; however, the mechanisms underlying such impairment remain incompletely understood. In the present study, a dextran sulfate sodium-induced colitis model was used to investigate how the inflammatory environment damages the intestinal mucosa. The results demonstrated that colitogenic ambiance enhances intestinal epithelial cell death, delays epithelial cell proliferation, and exacerbates mucosal erosion. Unexpectedly, this work identified a previously unrecognized role for Notch signaling in mediating these effects. Specifically, the colitogenic milieu reduces Notch/mechanistic target of rapamycin complex 1 (mTORC1)-mediated intestinal epithelial cell proliferation to promote goblet cell differentiation. Chemical activation of Notch signaling stimulated intestinal epithelial cell proliferation and reduced goblet cell differentiation in the colitic mucosa, further aggravating mucosal damage. Conversely, inhibition of Notch or mTORC1 signaling during mucosal repair reduced intestinal epithelial cell proliferation and enhanced goblet cell differentiation, corroborating the implication of Notch and mTORC1 signaling in both processes. Collectively, these findings uncover a context-dependent role for the Notch-mechanistic target of rapamycin axis in regulating intestinal epithelial cell proliferation and differentiation in the colitic mucosa and suggest that its targeted modulation may hold therapeutic potential in inflammatory bowel diseases.

Animals↗
PubMed2026

Single-cell transcriptomics reveals that air-liquid interface culture promotes goblet cell differentiation and inhibits glycolysis in organoid cell monolayers.

Faithfully recapitulating the cellular heterogeneity of the intestinal epithelium is essential when using organoid models. Air-liquid interface (ALI) culture has been shown to promote secretory cell differentiation, but its impact on gene expression in each epithelial cell type remains unclear. In this study, we used single-cell RNA sequencing (scRNA-seq) to characterize the cellular heterogeneity of rabbit cecum-derived organoid monolayers grown under immerged or ALI conditions. We then compared these organoid cell type-specific gene expression profiles to a scRNA-seq atlas of the rabbit cecal epithelium in vivo. We selected the rabbit model notably because, unlike mice, it possesses BEST4+ epithelial cells, a newly discovered subset of mature absorptive cells. Our analysis revealed a high degree of transcriptomic similarity between in vivo and organoid-derived stem and transit-amplifying cells. ALI culture markedly enhanced the differentiation of the secretory lineage, especially goblet cells, whose transcriptome closely resembled that of in vivo goblet cells. Furthermore, ALI was the only condition allowing the detection of enteroendocrine cells. BEST4+ cells, however, were absent from organoids in immerged or ALI conditions despite their presence in vivo. In addition, ALI culture led to a consistent downregulation of hypoxia and glycolysis-associated genes across all cell types, which suggests a metabolic shift likely driven by increased oxygen availability in ALI conditions. Cell-cell communication analyses further indicated that ALI more closely mirrored in vivo patterns than immerged condition. Altogether, these results demonstrate that ALI culture allows for better recapitulation of the in vivo cellular heterogeneity and molecular signatures of the intestinal epithelium.NEW & NOTEWORTHY Using single-cell RNA sequencing, this study shows that air-liquid interface (ALI) culture enhances secretory lineage differentiation of intestinal organoid cell monolayers and improves transcriptomic similarity to the native epithelium. ALI reduced hypoxia-associated gene expression and better recapitulates in vivo-like cell-cell interactions, supporting its value for modeling intestinal epithelial heterogeneity in organoids.

Animals↗
PubMed1979

Purification of human intestinal goblet cell antigen (GOA), its immunohistological demonstration in the intestine and in mucus producing gastrointestinal adenocarcinomas.

Goblet cell antigen (GOA) was purified from gastric signet ring cell carcinoma. It was immunogenic and was used to produced antisera which stained goblet cells of the small and large intestine and of intestinalized gastric mucosa by indirect immunological methods. Various types of gastric and colonic cancer contained GOA. These findings demonstrate a histiogenic relationship between intestinal goblet cells, various gastrointestinal cancers and associated premalignant conditions.

Adenocarcinoma↗
PubMed1975

Calcium binding to intestinal goblet cell mucin.

1. Binding of Ca-2+ to goblet cell mucin of rat small intestine was studied using equilibrium dialysis against 0.01 M Tris/HCl buffer (pH 7.4) and tracer amounts of 45-CaCl2. Binding was found to reach saturation at a Ca free -2+ concentration of 0.1--1.0 mM, to be independent of temperature (4-37 degrees C), and to increase with increasing pH (5.0-8.7). At low concentrations of Ca free -2+ (smaller than 0.03 mM) the binding curve was sigmoidal, suggesting positive cooperativity of binding sites and a possible change in the tertiary structure of the mucin. Binding was markedly reduced, and sigmoidicity abolished, by removal of sialic acid from the mucin, or by adding 0.14 M NaCl to the dialysis medium. This latter finding suggests that, in vivo, other cations would compete for Ca-2+ binding ligands. 2. Under conditions mimicking those used for binding studies, CaCl2 (10- minus 5 M) was found to cause a small increase (0.03 units) in the absorbance of mucin solutions, especially in the ultraviolet region, possibly indicating increased light scattering. No change in the solubility of the mucin was observed after the addition of CaCl2 (10- minus 6-10- minus 4 M). A significant decrease in viscosity of the mucin was noted, however, with the addition of CaCl2 (10- minus 6-10- minus 2 M). Together with the binding data, these findings suggested that during binding, Ca-2+ combines with negative charges on goblet cell mucin (especially those of sialic acid carboxyl groups) and induces contraction or folding of the macromolecule which promotes cooperative cation binding. No evidence was obtained to suggest that CaCl2 caused precipitation, polymerization or gelation of the mucin in 0.01 M Tris/HCl.?

Animals↗
PubMed1979

Intestinal goblet cell mucus release. II. In vivo stimulation by antigen in the immunized rat.

We previously reported that the infusion of certain soluble immune complexes stimulated mucus release from the rat small intestine in vivo. The present studies sought to evaluate the response of the intestine of normal and immunized rats to the infusion of antigen alone. One hour after the intraduodenal infusion of antigen, small intestinal washings were obtained and analyzed for the presence of 35S-labeled, high m.w. glycoprotein of goblet cell origin. The amount of goblet cell glycoprotein released was estimated from the radioactivity present in the void volume of a Sepharose 4B gel filtration column. The release of goblet cell mucus was enhanced by antigen stimulation in orally immunized animals. The discharge of goblet cell mucus was not increased after antigen infusion in animals immunized by the i.p. route despite the induction of high levels of serum antibody. The inability to demonstrate release of mucus after antigen challenge in systemically immunized rats suggests that the amount or the type(s) of antibody required at the mucosal surface is produced only after oral immunization.

Animals↗
PubMed1977

Quantitative differences in goblet cells in the tracheal epithelium of male and female rats.

The cause of higher morbidity rates among men than women from diseases of the respiratory tract may be complex and involve several factors, one of which may be structural differences within airways. In an attempt to determine whether such differences exist, 13 male rats and 45 female rats (15 each in proestrous, estrous, and diestrous) were evaluated. Tracheas were fixed in situ by injection of a mixture of formaldehyde and glutaraldehyde. Longitudinal sections cut from paraffin blocks were stained and combined alcian blue (pH 2.5) and periodic acid-Schiff stain to count goblet cells, or with Weigert's hematoxylin and eosin stain to determine epithelial thickness. In the normal rat trachea, cells containing periodic acid-Schiff-positive granules at their apices constituted a major population of goblet cells. The number of these goblet cells present in the trachea was greater in female than in male rats at each stage of the estrous cycle, with the values in the diestrous females closest to the value in the males. Among females, estrous and proestrous rats contained significantly more of these goblet cells than did diestrous animals. The tracheal epithelium of male rats was significantly thicker than that of female rats. These studies demonstrate that the tracheal epithelium differs in male and female rats and varies in the female rat as a function of hormonal cycles. These results raise the question of whether similar differences might be important in the pathogenesis of human disease, and they deserve further clarification in man.

Animals↗
PubMed1976

[Goblet cells in nasal allergy (author's transl)].

After staining of the whole nasal mucosa from a patient with nasal allergy by the PAS-alcian-blue whole mount method, the density of goblet cells was determined quantitatively. In most sections of the nasal septum and of the inferior and middle nasal concha, the density of goblet cells was found to be considerably lower than in normal subjects. While in some sections, in particular posteriorly, the density was the same as in normal subjects, it was nowhere higher. The low density may be due to metaplastic changes of the epithelium which had thickened and lost ciliary cells. Previous statements that the density of goblet cells is increased in nasal allergy could not be confirmed quantitatively, but investigation of larger materials is called for before any conclusions may be drawn.

Adult↗
PubMed1978

Alcian blue staining intestinal goblet cell antigen (GOA): a marker for gastric signet ring cell and colonic colloidal carcinoma.

An Alcian blue staining, perchloric acid-soluble, antigenic acidic mucosubstance (GOA) was purified from human gastric signet ring cell carcinoma with DEAE-cellulose chromatography, Seqhadex G-200 and preparative polycrylamide gel electrophoresis. Specific antisera were raised which reacted in indirect immunoenzyme histology with normal goblet cells of the small and large intestine and with goblet cells of intestinalized gastric mucosa. In surgical resection specimens of the stomach (n = 100) and of the colon (n = 19) 3 gastric signet ring cell carcinomas and 3 colonic colloidal adenocarcinomas stained for GOA, demonstrating an immunochemical relationship with normal intestinal goblet cells.

Adenocarcinoma, Mucinous↗
PubMed2025

Cetobacterium somerae ZNN-1 promotes goblet cell differentiation through glutamine-mediated Notch signaling suppression.

INTRODUCTION: The gut microbiota acts as a crucial mediator in the interaction between the diet components and the host metabolism. However, the molecular mechanism by which the gut microbiota adapts to dietary components and subsequently regulates host physiological responses remains unclear. OBJECTIVES: This study aimed to investigate the response of gut microbiota to a plant-based protein diet (soybean meal, SM) and the effects of gut microbiota on host intestinal barrier function, along with the underlying mechanisms in a fish model. METHODS: Histopathological examination, and transepithelial electrical resistance test were used to evaluate the effects of Cetobacterium somerae on intestinal barrier function. Potential molecular mechanisms were validated by integrating whole-genome sequencing, microbiota composition sequencing, transcriptomics, and metabolomics, and utilizing in vitro cell models and mouse-derived organoid models. RESULTS: The results revealed that the SM diet significantly increased the abundance of Cetobacterium somerae in fish. Administration of C.somerae ZNN-1, a dominant strain isolated from the intestine of fish fed with the SM diet, enhanced the intestinal barrier function, particularly increasing the number of goblet cells in the intestine. Whole genome analysis of C. somerae ZNN-1 showed carbohydrate metabolism-associated genes were the most abundant in its metabolic modules. C.somerae ZNN-1 supplementation significantly inhibited the Notch signaling pathway in fish intestine. Metabolomics analysis revealed that administration of C.somerae ZNN-1 increased the glutamine level in fish gut. In vitro experiments demonstrated that glutamine regulated the differentiation of goblet cell by inhibiting the Notch signaling pathway in both human intestinal epithelial cell model and mouse intestinal organoid model. CONCLUSION: C. somerae served as a key bacterium adapted to soybean meal-derived carbohydrates, and it promoted goblet cell differentiation by inhibiting the Notch pathway. This study provides a new perspective for unraveling the interaction mechanisms among diet components, intestinal microbiota and host health.

Animals↗
PubMed1979

Intestinal goblet cell differentiation in Nippostrongylus-infected rats after transfer of fractionated thoracic duct lymphocytes.

Adoptive immunization of Nippostrongylus-infected rats augmented goblet cell differentiation in the intestinal epithelium. After fractionation of immune thoracic duct lymphocytes (TDL), cells lacking surface immunoglobulin (sIg-) were the most potent stimulators of goblet cell differentiation. Moreover, TDL drained from rats harbouring a primary infection were more effective than TDL from hyperimmune rats.

Animals↗
PubMed1977

[Electron microscopic of mucopolysaccharides in the goblet cells of the bronchial mucosa].

Author using various methods of fixation and electronmicroscopical histochemistry investigated the detectability of mucopolysaccharides in the goblet cells of the bronchial mucosa obtained by biopsy. Results show, that the method of choice of the detection of glycosaminoglycans in the goblet cells is the PA-Silver technique carried out after fixation with solution of glutaraldehyde only. This technique-- when appropriate control is used--seems to be specific.

Biopsy↗
PubMed1979

Goblet cell carcinoid tumor of the appendix. Report of five cases and review of the literature.

Five cases of goblet cell carcinoid tumor of the appendix showed characteristic histologic features that justified classification of these lesions as mucinous variants of carcinoid tumor. The tumor has low-grade malignancy, and metastases are uncommon. Resemblance to mucinous adenocarcinoma of the appendix is striking, and the features that help to differentiate the two lesions are delineated.

Adenocarcinoma, Mucinous↗
PubMed1979

Radioimmunoassay of human intestinal goblet cell mucin. Investigation of mucus from different organs and species.

We have developed a double-antibody radioimmunoassay for the quantitative measurement of human goblet cell mucin (GCM) in order to study intestinal mucus in human and other species. The assay used 3H-labeled mucin as the antigen, rabbit antisera, and sheep anti-rabbit IgG antisera as the second antibody. A number of applications of the assay were investigated. A survey of human tissues revealed that mucins of the rectum, colon, and small intestine had identical affinity for the rabbit antibody, whereas lung eyelid conjunctiva, esophagus, and stomach reacted less strongly. GCM concentration ranged from 1.9 to 14 microgram mucin protein/mg tissue protein in the small and large intestine, respectively. The radioimmunoassay was also found to be useful as a marker during the isolation of GCM from human ileal extracts, where it indicated that a 10,000-fold purification had been achieved. Antigenic determinants of the mucin did not rely upon ABH blood group-specific terminal sugars in oligosaccharide chains. A comparison of mucins among various species revealed a partial species specificity of the GCM antibody. Human GCM cross-reacted with dog, monkey, and rabbit mucins, but not with mucins of rat, pig, toad, and oyster. Organ distributions of cross-reactive mucins in rabbit tissues indicated a pattern that was qualitatively similar to that seen in human tissues. Possible implications of these findings for autoimmune diseases are briefly discussed.

Animals↗
PubMed1976

An ultrastructural and histochemical study of the mucous membrane adjacent to and remote from carcinoma of the colon.

The material was taken from surgical specimens of large intestine resected for carcinoma. A parallel study by electron microscopy and mucin histochemistry was made on fragments from "transitional" (TR) mucosa (adjacent to carcinoma) and "normal" (N) mucosa (remote from the tumor). These were compared with similar studies on the mucosa from control individuals. Histochemically, the "TR" mucosa shows an increase in sialomucins as compared with the "N" and control mucosae, where sulphomucins normally predominate. At the ultrastructural level, the "TR" mucosa is characterized by the following changes which may precede the histochemical variations: A) alteration in the relative proportions of the different cell types, along the crypt, with a persistence of immature and intermediate cells at higher levels of the crypt than in the control. Mature absorptive cells are fewer and bear an inverse relationship to intermediate cells. Goblet cells are increased in number and size. B) Appearance of electron-dense bodies 0.15-0.3mum in in diameter and membrane limited. C) An elaborated and enlarged Golgi zone showing increased secretory activity. It is suggested that mucin and ultrastructural changes described in the "TR" zone may indicate a failure in the normal process of cell differentiation along the crypt. Previous work7,8 further suggests that the mucin changes may be primary, reflecting a cellular response to unknown stimuli (i.e., carcinogens) rather than a local secondary effect of tumor growth. As for the ultrastructural features in the "TR" mucosa, the secondary effect cannot be excluded at the present.

Carcinoma↗