Moluccella laevis lectin, a marker for cellular differentiation programs in mouse gut epithelium

Abstract

We have assembled a system for testing the hypothesis that changes in glycoconjugate production represent markers for defining developmental, spatial, and environmental influences on the proliferation and differentiation programs of various mouse gut epithelial cell lineages. Multilabel immunohistochemical methods were used to survey the interactions of purified lectins with 1) normal fetal, neonatal, and adult FVB/N mouse gut, 2) gastric and intestinal isografts harvested at various developmental stages, and 3) transgenic mouse models of intestinal epithelial cell hyperplasia, dysplasia, and/or neoplasia. As a demonstration of the system's utility, we used the recently purified, alpha-N-acetyl-D-galactosamine-specific, Moluccella laevis lectin (MLL). In the adult FVB/N mouse stomach, MLL only recognizes glycoconjugates produced by a population of nonproliferating neck and prezymogenic cells that occupy a pivotal point in the complex, migration-associated differentiation program of the zymogenic cell lineage. In the developing FVB/N stomach, MLL binds to members of the zymogenic and pit lineages even before morphogenesis of gastric units is completed. Expression of MLL epitopes in pit cells is restricted to the period before the gastric epithelium has completed its morphoregulatory program. Analysis of gastric isografts indicates that these lineage- and developmental stage-specific patterns of glycoconjugate accumulation are not influenced by normal luminal contents. In the adult FVB/N intestine, MLL binding can be used to operationally define variations in the differentiation programs of 1) members of the enteroendocrine and goblet cell lineages during their migration along the crypt-to-villus axis and 2) cells comprising the follicle-associated epithelium overlying Peyer's patches. Accumulation of MLL epitopes in villus-associated enterocytes does not appear to be affected when these cells are induced to reenter the cell cycle by simian virus 40 large T antigen (SV40 TAg). MLL reactivity is not diminished when enterocytes begin to dedifferentiate as a result of production of SV40 TAg, human K-rasVal12, and a dominant negative human p53 mutant. The lack of change in MLL binding properties may reflect the brief residence time of enterocytes on the villus. These results indicate that glycoconjugate production represents a very useful tool for studying gut epithelial cell biology. Preliminary studies suggest that this is also true in the human gut.

Epithelial BMP signaling is required for proper specification of epithelial cell lineages and gastric endocrine cells

“…Indeed there is a probability where misspecification of these cells might lead to a gain of expression in UEA1 lectin. Under normal conditions, this specific lectin is expressed exclusively by the mucous surface cell in the mouse gastric epithelium (9). Nevertheless, future development of specific markers for the different gastric precursor cells should allow the accurate evaluation of the exact role for BMP signaling on the specification of these cell subpopulations.…”

Gastrin stimulates the growth of gastric pit cell precursors by inducing its own receptors

“…An LSAB2/HRP staining kit (DAKO Japan) was used as the secondary antibody reaction system. Gastric pit cells were visualized using fluorescein-5-isothiocyanatelabeled CTB lectin (Sigma, St Louis, MO) binding as previously reported (6,7).…”

Diphtheria Toxin-mediated Ablation of Parietal Cells in the Stomach of Transgenic Mice

“…Anguilla anguilla agglutinin (15)(16)(17). Histochemical studies with these lectins suggested that there was an increase in pit cell number in the zymogenic gastric units of transgenic mice (e.g.…”