Fluorochrome-coupled lectins reveal distinct cellular domains in human epidermis.

Abstract

The distribution of saccharide moieties in human interfollicular epidermis was studied with fluorochrome-coupled lectins. In frozen sections Concanavalin A (Con A), Lens culinaris agglutinin (LCA), Ricinus communis agglutinin I (RCAI), and wheat germ agglutinin (WGA) stained intensively both dermis and viable epidermal cell layers, whereas peanut agglutinin (PNA) bound only to living epidermal cell layers. Ulex europaeus agglutinin I (UEAI) bound to dermal endothelial cells and upper cell layers of the epidermis but left the basal cell layer unstained. Dolichos biflorus agglutinin (DBA) bound only to basal epidermal cells, whereas both soybean agglutinin (SBA) and Helix pomatia agglutinin (HPA) showed strong binding to the spinous and granular cell layers. On routinely processed paraffin sections, a distinctly different staining pattern was seen with many lectins, and to reveal the binding of some lectins a pretreatment with protease was required. All keratin-positive cells in human epidermal cell suspensions, obtained with the suction blister method, bound PNA, whereas only a fraction of the keratinocytes bound either DBA or UEAI. Such a difference in lectin binding pattern was also seen in epidermal cell cultures both immediately after attachment and in organized cell colonies. This suggests that in addition to basal cells, more differentiated epidermal cells from the spinous cell layer are also able to adhere and spread in culture conditions. Gel electrophoretic analysis of the lectin-binding glycoproteins in detergent extracts of metabolically labeled primary keratinocyte cultures revealed that the lectins recognized both distinct and shared glycoproteins. A much different lectin binding pattern was seen in embryonic human skin: fetal epidermis did not show any binding of DBA, whereas UEAI showed diffuse binding to all cell layers but gave a bright staining of dermal endothelial cells. This was in contrast to staining results obtained with a monoclonal cytokeratin antibody, which showed the presence of a distinct basal cell layer in fetal epidermis also. The results indicate that expression of saccharide moieties in human epidermal keratinocytes is related to the stage of cellular differentiation, different cell layers expressing different terminal saccharide moieties. The results also suggest that the emergence of a mature cell surface glycoconjugate pattern in human epidermis is preceded by the acquisition of cell layer-specific, differential keratin expression.

Binding of the blood group‐reactive lectins to human adult kidney specimens

“…Although some studies have reported that the DBA conjugates, with a nominal specificity for blood group AI, would react with endothelia in blood group A specimens (Hsu and Ree, 1983;Schaumburg-Lever et al, 19841, both our earlier studies (Virtanen et al, 1986;Hormia et al, 19881, those of others (Damjanov, 19871, Fig. 3. Reactivity of the GSA-I-B4 and SJA conjugates with human kidney specimens.…”

Light microscopic lectin histochemistry in aging mouse kidney: study of compositional changes in glycoconjugates.

“…Glycoconjugates are generally regarded as playing important roles in, e.g., the morphogenesis of organs, transformation of cells (5,19), and adhesion of ceh (3,29). VariouS lectin conjugates such as H'K-kctin (6,7,33,34), peroxrdase-lectin (23,26), biotinylated lectin (8,15,35), or colloid?l gold-kctin (20,21) h2R been developed U, demonstrate the localizltion o f glycoconjugates histochemically. Lectin histochemistry has been employed to characterize the glycoconjugates in adult kidney tissues at both light and electron microscopic levels.…”

Expression and Localization of N acetyl glucosamine in diabetic rats

“…Lektinler karbonhidrat bağlayan proteinlerdir, en az iki şeker bağlama bölgesi içerirler, enzimatik aktivite göstermezler ve insan, bakteri veya bitki kaynaklı olabilirler Genellikle çeşitli epitellerdeki glikokonjugat dağılımları 8,9 , gelişim ve farklılaşma üzerine çalışmalarda kullanılırlar 10 . Lektinlerle ilgili çok fazla sayıda çalışma olmasına rağmen diş eti dokusu, diyabet ve lektinler arasındaki ilişkiler hakkında çalışma yok denecek kadar azdır, özellikle yapılan literatür araştırmalarında diyabetik hastaların diş eti dokusunun lektinlerle olan ilişkilerine dair bir çalışmaya rastlanmamıştır.…”