Skip to main content
Log in

Subcellular characterization of glycoproteins in the principal cells of human gallbladder

A lectin cytochemical study

  • Original Paper
  • Published:
Histochemistry Aims and scope Submit manuscript

Abstract

Gallbladder mucus is mainly composed of glycoproteins, which seem to play a critical role in cholesterol nucleation during gallstone formation. The biosynthetic pathway and sequential processing as well as the characterization of the oligosaccharide sidechains of human gallbladder secretory glycoproteins have not been completely defined. The aim of the present study is the subcellular characterization of the glycoproteins in the principal cells of human gallbladder. Principal cells of normal human gallbladder were studied by means of a variety of cytochemical techniques, including lectin histochemistry, enzyme and chemical treatments, immunocytochemistry and lectin-gold technology. Fucose, galactose, N-acetylglucosamine, N-acetylgalactosamine and N-acetylneuraminic acid residues were detected in mucous granules, Golgi apparatus and apical membrane of principal cells. Mannose residues were only observed in dense bodies. Oligosaccharide side-chains of the glycoproteins contained in the biliary mucus are synthesized in the Golgi apparatus of the principal cells of the gallbladder epithelium and are also contained in the mucous granules of these cells. Terminal N-acetylneuraminic acid(α2-3)galactose(β1-3)N-acetylgalactosamine, N-acetylneuraminic acid(α2-3)galactose(β1-4)N-acetylglucosamine and galactose(β1-4)N-acetylglucosamine sequences are contained in the oligosaccharide chains of gallbladder mucus glycoproteins. The dense bodies detected in the cytoplasm of the principal cells contained N-linked glycoproteins. Mucin-type O-linked glycoproteins were the main components of the mucous granules although some N-linked chains were also detected.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Baenziger JU, Fiete D (1979) Structural determinants of Ricinus communis agglutinin and toxin specificity for oligosaccharides. J Biol Chem 254:9795–9799

    Google Scholar 

  • Bouchier IAD, Cooperband SR, El Kodsi BM (1965) Mucous substances and viscosity of normal and pathologic bile. Gastroenterology 49:343–353

    Google Scholar 

  • Brewer CF, Bhattacharyya L (1988) Concanavalin A interactions with asparagine-linked glycopeptides. The mechanism of binding of oligomannose bisected hybrid and complex type carbohydrates. Glycoconjugate J 5:159–173

    Google Scholar 

  • Carlemalm E, Garavito M, Villiger W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc 126:123–143

    Google Scholar 

  • Castells MT, Ballesta J, Pastor LM, Madrid JF, Marin JA (1990) Histochemical characterization of glycoconjugates in the epithelium of the extrapulmonary airways of several vertebrates. Histochem J 22:24–35

    Google Scholar 

  • Castells MT, Ballesta J, Madrid JF, Avilés M, Martínez-Menárguez JA (1991) Characterization of glycoconjugates in developing rat respiratory system by means of conventional and lectin histochemistry. Histochemistry 95:419–426

    Google Scholar 

  • Castells MT, Ballesta J, Madrid JF, Martínez-Menárguez JA, Avilés M (1992) Ultrastructural location of glycoconjugates in human bronchial glands: the subcellular organization of N- and O-linked oligosaccharide chains. J Histochem Cytochem 40:265–274

    Google Scholar 

  • Debray H, Decout D, Strecker G, Spik G, Montreuil J (1981) Specificity of twelve lectins towards oligosaccharides and glycopeptides related to N-glycosylproteins. Eur J Biol Chem 117:41–55

    Google Scholar 

  • Egea G, Goldstein IJ, Roth J (1989) Light and electron microscopic detection of (3 Gal β 1,4 GlcNAc β1) sequences in asparagine-linked oligosaccharides with the Datura stramonium lectin. Histochemistry 92:515–522

    Google Scholar 

  • Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nature (Phys Sci) 241:20–22

    Google Scholar 

  • Freston JW, Bouchier IAD, Newman J (1969) Biliary mucous substances in dihydrocholesterol-induced cholelithiasis. Gastroenterology 67:670–680

    Google Scholar 

  • Gallinger S, Taylor RD, Harvey PRC, Petrunka CN, Strasberg S (1985) Effects of mucous glycoprotein on nucleation time in human bile. Gastroenterology 98:648–658

    Google Scholar 

  • Geoghegan WD, Ackerman GA (1977) Adsorbtion of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem Cytochem 25:1187–1200

    Google Scholar 

  • Gilloteaux J, Pomerants B, Kelly TR (1989) Human gallbladder mucosa ultrastructure: evidence of intraepithelial nerve structures. Am J Anat 184:321–333

    Google Scholar 

  • Goldstein IJ, Hayes CE (1978) The lectins: carbohydrate-binding proteins of plants and animal. Adv Carbohydr Chem Biochem 35:127–140

    Google Scholar 

  • Green ED, Brodbeck RM, Baenziger JU (1987) Lectin affinity high-performance liquid chromatography. J Biol Chem 262:12030–12039

    Google Scholar 

  • Hacker GW, Grimelius L, Danscher G, Bernatzky G, Muss W, Adam H, Thurner J (1988) Silver acetate autometallography: an alternative enhancement technique for immunogold-silver staining (IGSS) and silver amplification of gold, silver, mercury and zinc in tissues. J Histotechnol 11:213

    Google Scholar 

  • Hayward AF (1968) The structure of the gall bladder epithelium. Int Rev Gen Exp Zool Lond 3:205–239

    Google Scholar 

  • Karayannopoulou G, Damjanov I (1987) Lectin binding sites in the human gallbladder and cystic duct. Histochemistry 88:75–83

    Google Scholar 

  • Knibbs RN, Goldstein IJ, Ratcliffe RM, Shibuya N (1991) Characterization of the carbohydrate binding specificity of the leukoagglutinin lectin from Maackia amurensis. Comparison with other sialic acid-specific lectins. J Biol Chem 266:83–88

    Google Scholar 

  • La Mont JT, Smith BF, Moore JRL (1984) Role of gallbladder mucin in pathophysiology of gallstones. Hepatology 4:51S-56S

    Google Scholar 

  • Laboisse C, Bogomoletz WV (1989) Les mucines: des glycoprotéines en quête de reconnaissance. Ann Pathol 9:175–181

    Google Scholar 

  • Lee SP, Lim TH, Scott AJ (1979) Carbohydrate moieties of glycoproteins in human hepatic and gallbladder bile, gallbladder mucosa and gallstones. Clin Sci Mol Med 56:533–538

    Google Scholar 

  • Lee SP, La Mont JT, Carey MC (1981) The role of gallbladder mucus hypersecretion in the evolution of cholesterol gallstones: studies in the prairie dog. J Clin Invest 67:1712–1723

    Google Scholar 

  • Levy PF, Smith BF, La Mont JT (1984) Human gallbladder mucin accelerates in vitro nucleation of cholesterol in artificial bile. Gastroenterology 87:270–275

    Google Scholar 

  • Lotan R, Skutelsky E, Danon D, Sharon N (1975) The purification, composition, and specificity of the anti-T lectin from peanut (Arachis hypogaea). J Biol Chem 250:8518–8523

    Google Scholar 

  • Luciano L, Reale E (1990) The human gallbladder. In: Riva A, Motta PM (eds) Ultrastructure of the extraparietal glands of the digestive tract. Kluver Academic Publishers, Boston, pp 231–248

    Google Scholar 

  • Luciano L, Wermbter G, Reale E (1973) Die Feinstruktur der Gallenblase und der Gallengänge. III. Beobachtungen an gefriergeätzten Präparaten der Gallenblase der Maus unter besonderer Berücksichtigung der dichten Körper und des Verbindungskomplexes. Cytobiologie 7:76–78

    Google Scholar 

  • Madrid JF, Ballesta J, Galera T, Castells MT, Pérez-Tomás R (1989a) Histochemistry of glycoconjugates in the gallbladder epithelium of ten animal species. Histochemistry 91:437–443

    Google Scholar 

  • Madrid JF, Ballesta J, Castells MT, Marín JA, Pastor LM (1989b) Characterization of glycoconjugates in the intestinal mucosa of vertebrates by means of lectin histochemistry. Acta Histochem Cytochem 22:1–14

    Google Scholar 

  • Madrid JF, Ballesta J, Castells MT, Hernández F (1990) Glycoconjugate distribution in the human fundic mucosa revealed by lectin- and glycoprotein-gold cytochemistry. Histochemistry 95:179–187

    Google Scholar 

  • Mandal C, Mandal C (1990) Sialic acid binding lectins. Experientia 46:433–441

    Google Scholar 

  • Martínez-Menárguez JA, Avilés M, Castells MT, Madrid JF, Ballesta J (1992) Cytochemical characterization of the oligosaccharide biosynthesis of rat acrosomal glycoproteins. Histochem J 24:503

    Google Scholar 

  • Martínez-Menárguez JA, Avilés M, Madrid JF, Castells MT, Ballesta J (1993) Glycosylation in Golgi apparatus of early spermatids of rat. A high resolution lectin cytochemical study. Eur J Cell Biol 61:21–33

    Google Scholar 

  • Nevalainen TJ, Dean S, Jokelainen PT, Laitio M (1976) Lamellated osmiophilic bodies in mouse gallbladder epithelium. Acta Anat 96:486–496

    Google Scholar 

  • Osawa T, Tsuji T (1987) Fractionation and structural assessment of oligosaccharides and glycopeptides by use of immobilized lectins. Annu Rev Biochem 56:21–42

    Google Scholar 

  • Pearson JP, Kaura R, Taylor W, Allen A (1982) The composition and polymeric structure of mucus glycoprotein from human gallbladder bile. Biochem Biophys Acta 706:221–228

    Google Scholar 

  • Pereira MEA, Kabat EA (1974) Blood group specificity of the lectin Lotus tetragonolobus. Ann NY Acad Sci 234:301–305

    Google Scholar 

  • Piller V, Piller F, Cartron JP (1990) Comparison of the carbohydrate-binding specificities of seven N-acetyl-d-galactosamine-recognizing lectins. Eur J Biochem 191:461–466

    Google Scholar 

  • Roberts GP (1977) Histochemical detection of sialic acid residues using periodate oxidation. Histochem J 9:97–102

    Google Scholar 

  • Roth J (1983a) Applications of immunocolloids in light microscopy. II. Demonstration of lectin-binding sites in paraffin sections by the use of lectin-gold or glycoprotein gold complexes. J Histochem Cytochem 31:547–552

    Google Scholar 

  • Roth J (1983b) Application of lectin-gold complexes for electron microscopic localization of glycoconjugates on thin sections. J Histochem Cytochem 31:987–999

    Google Scholar 

  • Roth J (1984) Cytochemical localization of terminal N-acetyl-d-galactosamine residues in cellular compartments of intestinal goblet cells: implications for the topology of O-glycosylation. J Cell Biol 98:399–406

    Google Scholar 

  • Roth J (1987) Subcellular organization of glycosylation in mammalian cells. Biochim Biophys Acta 906:405–436

    Google Scholar 

  • Roth J, Lucocq JM, Charest PM (1984) Light and electron microscopic demonstration of sialic acid residues with the lectin from Limax flavus: a cytochemical affinity technique with the use of fetuin-gold complexes. J Histochem Cytochem 32:1167–1176

    Google Scholar 

  • Sata T, Lackie PM, Taatjes DJ, Peumans W, Roth J (1989) Detection of Neu5Ac (α2,3)Gal(β1,4)GlcNAc sequences with the leukoagglutinin from Maackia amurensis: light and electron microscopic demonstration of differential tissue expression of terminal sialic acid in α2,3 and α2,6-linkage. J Histochem Cytochem 37:1577–1588

    Google Scholar 

  • Sata T, Zuber C, Roth J (1990) Lectin-digoxigenin conjugates: a new hapten system for glycoconjugate cytochemistry. Histochemistry 94:1–11

    Google Scholar 

  • Schauer R (1982) Chemistry, metabolism and biological functions of sialic acid. Adv Carbohydr Chem Biochem 40:131–234

    Google Scholar 

  • Shibuya N, Goldstein IJ, Broekaert WF, Nsimba-Lubaki M, Peeters B, Peumans WJ (1987) The elderberry (Sambucus nigra L.) bark lectin recognizes the Neu5Ac (α2-6)Gal/GalNAc sequence. J Biol Chem 255:9719–9723

    Google Scholar 

  • Shibuya N, Goldstein IJ, Van Damme EJM, Peumans WJ (1988) Binding properties of a mannose specific lectin from the snowdrop (Galanthus nivalis) bulb. J Biol Chem 263:728–734

    Google Scholar 

  • Slot JW, Geuze HJ (1985) A new method of preparing gold probes for multiple-labelling cytochemistry. Eur J Cell Biol 38:87–93

    Google Scholar 

  • Smith BF (1987) Human gallbladder mucin binds biliary lipids and promotes cholesterol crystal nucleation in model bile. J Lipid Res 28:1088–1097

    Google Scholar 

  • Smith BF, La Mont JT (1984) Hydrophobic binding properties of bovine gallbladder mucin. J Biol Chem 259:12170–12177

    Google Scholar 

  • Smith BF, Peetermans JA, Tanaka T, La Mont T (1989) Subunit interactions and physical properties of bovine gallbladder mucin. Gastroenterology 97:179–187

    Google Scholar 

  • Sugii S, Kabat EA (1982) Further immunochemical studies on the combining sites of Lotus tetragonolobus and Ulex europeaus I and II lectins. Carbohydr Res 99:99–101

    Google Scholar 

  • Taatjes DJ, Roth J, Peumans W, Goldstein IJ (1988) Elderberry bark lectin-gold techniques for the detection of Neu5Ac (α2,6)Gal/GalNAc sequences: applications and limitations. Histochem J 20:478–490

    Google Scholar 

  • Von Figura K, Hasilik A (1986) Lysosomal enzymes and their receptors. Annu Rev Biochem 55:167–193

    Google Scholar 

  • Wahlin T (1979) Histochemical analysis of mucosubstances and cytochemical studies of the Golgi region and the secretory granules of the normal gerbil gallbladder principal cells. Acta Anat 103:468–476

    Google Scholar 

  • Wu AM, Sugii S (1991) Coding and classification of d-galactose, N-acetyl-d-galactosamine, and β-d-Galp[1->3(4)]-β-d-Glcp-NAc, specificities of applied lectins. Carbohydr Res 213:127–143

    Google Scholar 

  • Yamashita K, Kochibe N, Ohkura T, Ueda I, Kobata A (1985) Fractionation of l-fucose-containing oligosaccharides on immobilized Aleuria aurantia lectin. J Biol Chem 260:4688–4693

    Google Scholar 

  • Yamashita K, Tokani K, Ohkura T, Takasaki T, Goldstein IJ, Kobata A (1987) Carbohydrate binding properties of complextype oligosaccharides on immobilized Datura stramonium lectin. J Biol Chem 262:1602–1607

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Madrid, J.F., Castells, M.T., Martínez-Menárguez, J.A. et al. Subcellular characterization of glycoproteins in the principal cells of human gallbladder. Histochemistry 101, 195–204 (1994). https://doi.org/10.1007/BF00269544

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00269544

Keywords

Navigation