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Histochemical methods for characterizing secretory and cell surface sialoglycoconjugates.

B A Schulte, S S Spicer

    The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
    |May 1, 1985
    PubMed
    Summary
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    This study used lectin staining to map carbohydrate structures in rat, mouse, and hamster tissues. Peanut agglutinin (PNA) and Dolichos biflorus agglutinin (DBA) revealed specific sugar residues in various cell types and secretions.

    Area of Science:

    • Glycobiology and Histochemistry
    • Comparative Anatomy of Rodent Tissues

    Background:

    • Carbohydrate structures (glycoconjugates) play crucial roles in cellular functions and interactions.
    • Specific lectins, like Peanut Agglutinin (PNA) and Dolichos biflorus Agglutinin (DBA), are valuable tools for identifying terminal sugar residues.
    • Understanding the distribution of these glycans is essential for characterizing tissue-specific functions and species differences.

    Purpose of the Study:

    • To characterize the distribution of terminal beta-galactose and alpha-N-acetylgalactosamine residues in the trachea, sublingual gland, and pancreas of rats, mice, and hamsters.
    • To investigate the presence of penultimate sugars after sialic acid removal using enzymatic digestion.
    • To identify glycans with O-acetylated sialic acid side chains through periodate oxidation resistance.

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    Main Methods:

    • Paraffin-embedded sections of trachea, sublingual gland, and pancreas from rats, mice, and hamsters were stained with PNA or DBA conjugated to horseradish peroxidase.
    • Enzymatic removal of sialic acid (sialidase digestion) and periodate oxidation were performed on adjacent sections prior to lectin staining.
    • Microscopic examination documented lectin binding sites to identify specific carbohydrate structures and their locations.

    Main Results:

    • PNA binding indicated terminal beta-galactose in various secretions and cell membranes, including mouse tracheal serous cells and pancreatic acinar cells.
    • Sialidase digestion revealed penultimate beta-galactose in mucous cell secretions and tracheal epithelium glycocalyx, and PNA affinity in endothelium and serous cells.
    • DBA binding identified terminal alpha-N-acetylgalactosamine in mucous cell secretions, tracheal epithelium, and glandular serous cells across species, with variations noted.

    Conclusions:

    • The study successfully mapped the distribution of specific terminal and penultimate carbohydrate residues in key rodent organs using PNA and DBA lectins.
    • Enzymatic and oxidative treatments provided insights into the presence of sialic acids, including O-acetylated forms, and their linkage to underlying sugars.
    • Distinct patterns of glycoconjugate expression were observed among species and tissue types, highlighting their potential functional significance.