Differentiation-Dependent Glycosylation of Cells in Squamous Cell Epithelia Detected by a Mammalian Lectin

Abstract

The squamous stratified epithelia contain a proliferative (harboring mitotic activity) and a differentiating compartment. Due to the potential of protein-carbohydrate interactions to regulate cellular activities we introduced a mammalian lectin to cyto- and histochemical analysis. We answer the questions of whether and to what extent this new probe can pinpoint differentiation-dependent glycosylation changes in sections and in culture of keratinocytes. Material and Methods: Purification and labeling enabled monitoring of galectin-3 reactivity in frozen sections of human and pig epidermis and basal cell carcinomas as well as in culture of keratinocytes. The staining pattern of the lectin was correlated with the staining profile of other cell markers including desmosomal proteins, β₁ integrin, and the proliferation marker Ki-67. The Dolichos biflorus agglutinin (DBA) sharing binding reactivity of galectin-3 to the A type histoblood group epitope was used for comparison. Results: Both lectins exhibit suprabasal binding. However, their profiles were not identical, substantiated by lack of coinhibition. Strong DBA reactivity was also observed in a limited number of basal layer cells, namely in cells without the expression of the proliferation marker Ki-67. Cultured mitotic epidermal cells have no reactivity for DBA. Presence of ligands for this plant lectin was connected with decreased positivity of nuclei for Ki-67 and the occurrence of ring-shaped nucleoli, micronucleoli or absence of nucleoli. Considering colocalization the pattern of galectin-3-binding sites coincided with the presence of desmosomal proteins such as desmoplakin-1 and desmoglein but not β₁ integrin, a potential ligand. Interestingly, studied basal cell carcinomas expressed no binding sites for galectin-3, while a limited number of cells were DBA-reactive. Conclusion: The expression of galectin-3-binding sites and also DBA-reactive glycoligands correlates with an increased level of differentiation and/or cessation of proliferation in the examined squamous stratified epithelia. Further application of tissue lectins for characterizing ligand expression and its modulation is an important step to reveal functional relevance.

Glycobiology of Head and Neck Squamous Epithelia and Carcinomas

“…P-cadherin, a molecule participating in the intercellular contact formation, is also expressed by basal cells only. As basal cells can proliferate, their nuclei are frequently positive for the proliferation marker Ki-67 and proliferating cell nuclear antigen [20]. The highly specific proteins connected with late terminal differentiation such as involucrin are expressed suprabasally and reflect the formation of the cornified envelope in the suprabasal keratinized cells.…”

“…This lectin and, similarly, accessible glycoligands reactive for Gal-3 are expressed by suprabasal cells [27,28]. No expression of proliferation markers such as Ki-67 is observed in cells positive for Gal-3 and its reactive glycoligands [20]. These cells do not express the ß 1 -chain of the integrin receptor either [29].…”

“…These cells do not express the ß 1 -chain of the integrin receptor either [29]. The Gal-3-reactive epitope expression is restricted to the intercellular contacts of suprabasal cells [20,28]. These binding sites precisely colocalize with the desmosomal proteins desmoplakin and desmoglein [20,28].…”

“…The Gal-3-reactive epitope expression is restricted to the intercellular contacts of suprabasal cells [20,28]. These binding sites precisely colocalize with the desmosomal proteins desmoplakin and desmoglein [20,28]. The cells recognized by Gal-3 strongly express cytokeratin type 10 [20].…”

“…These binding sites precisely colocalize with the desmosomal proteins desmoplakin and desmoglein [20,28]. The cells recognized by Gal-3 strongly express cytokeratin type 10 [20]. Labeled neoglycoligands with covalently linked defined saccharidic motifs reactive for Gal-3 recognize cells of the suprabasal layers of the studied epithelia.…”