Summary
Cell surface and cytoplasmic glycoconjugates were characterized in embryonic human explant cells (a transformed heteroploid line) cultured in iso-osmotic medium (0.137m NaCl) and in hyperosmotic medium (0.274m NaCl) for 10 days in order to study the changes induced in these compounds by hyperosmoticity. Cytochemical and ultracytochemical staining selective for glycoconjugates was carried out. The following results were obtained: (1) glycoproteins, glycosaminoglycans and glycolipids are present on the cell surface and in the cytoplasm of the explant cells; (2) lectin histochemistry combined with glycosidase digestion demonstrated the presence of the disaccharides fucose-N-acetylglucosamine and sialic acid-β-galactose as terminal sequences; (3) histophotometric evaluation of lectin labelling showed a noticeable decrease in histochemical reactivity of adapted cells; (4) plasma membrane cell coat decreased in adapted cells, which was emphasized by ultracytochemical reactions and a rearrangement of glycolipids in the cytoplasm.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baccarani Contri, M., Fornieri, C. &Pasquali Ronchetti, I. (1985) Elastin-proteoglycan association revealed by cytochemical methods.Connect. Tissue Res. 13, 237–49.
Ban, D. D. &Przelecka, A. (1981) Surface coat of plasma membrane of L-1210 lymphoid leukemia cells. A cytochemical study.Folia Histochem. Cytochem. 19, 3–10.
Bolognani, L., Bolognani Fantin, A. M., Castellana, C., Fuhrman Conti, A. M., Giuliani, A., Sabatini, M. A., Sanguini, C. L., Silvestri, M. G. &Tricoli, S. (1985) Enzymatic characterization of E.U.E. (embryonal human explants) cells adapted to hypertonic media.Acta histochem. 76, 1–11.
Bolognani, L., Bolognani Fantin, A. M., Fuhrman Conti, A. M., Gervaso, M. V. &Omodeo-Sale', M. F. (1978) Biochemical and histochemical features of human cultured cells (E.U.E.) adapted to hypertonic medium.Acta histochem. 62, 12–25.
Bolognani, L., Fuhrman Conti, A. M., Oberosler, R. &Giuliani De Bernardi, A. (1982) Investigation on the pattern of polar lipids extracted from different strains of cultured cells.Bas. appl. Histochem. 26, 217–22.
Bolognani, L., Fuhrman Conti, A. M. &Omodeo-Sale', M. F. (1976) Studies on polar lipids in human cultured cells adapted to hypertonic medium.Biochem. Exp. Biol. 12, 167–73.
Bolognani Fantin, A. M., Franchini, A., Fuhrman Conti, A. M., Corbascio, G. C. &Marchisio, P. C. (1988) Effects of hypertonic conditions on cytoskeletal and adhesion structures of EUE cells.Eur. J. Cell Biol. 47, 327–33.
Bolognani Fantin, A. M., Franchini, A., Sabatini, M. A. &Fuhrman Conti, A. M. (1987) Cytofunctional changes in E.U.E. cells adapated to hypertonic medium: scapning and transmission electron microscopy observations.Cell. Mol. Biol. 33, 51–61.
Bonucci, E. (1981)Manuale di Istochimica, pp. 154–4. Roma: Lombardo Ed.
Branny, J., Klein, A., Knycz, M., Drozd, CZ. &Pawelek, J. (1980) Preliminary results of the investigations of the cell surface membranes composition (proteins and sugars) in BHK-21, BHK-21/RSV and PR-RSH cell cultures.Folia Histochem. Cytochem. 18, 109–12.
Culp, L. A., Grimes, W. J. &Black, P. H. (1971) Contact-inhibited revertant cell lines isolated from SV 40-transformed cells.J. Cell Biol. 50, 682–90.
De Carli, L., Maio, J. J., Nuzzo, F. &Benerecetti, A. S. (1964) Cytogenetic studies with alkaline phosphatase in human heteroploid cells.Cold Spring Harbor Symp. Quant. Biol. 29, 223–31.
Faraggiana, T., Malchiodi, F., Prado, A. &Churg, J. (1982) Lectin-peroxidase conjugate reactivity in normal human kidney.J. Histochem. Cytochem. 30, 451–8.
Fuhrman Conti, A. M. (1979) Cytology and genetic features of mammalian cultured cells adapted to hypertonic medium.Bas. Appl. Histochem. suppl. 23, 51–7.
Graham, R. C. &Karnovsky, M. J. (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique.J. Histochem. Cytochem. 14, 291–302.
Kurth, P. D., Bustin, M. &Moudrianakis, E. N. (1979) Concanavalin A binds to puffs in polytene chromosomes.Nature 279, 448–50.
Luft, J. H. (1971) Ruthenium red and violet II. Fine structural localization in animal tissues.Anat. Rec. 171, 369–416.
Makita, A. &Seyama, Y. (1971) Alterations of Forsman-antigenic reactivity and of monosaccharide composition in plasma membrane from polyoma-transformed hamster cells.Biochim. Biophys. Acta 241, 403–11.
Manfredi Romanini, M. G., Fuhrman Conti, A. M. &Pellicciari, C. (1987) Effects of hypertonic medium on human cell growth: II. Heterogeneity of Feulgen-DNA content and nuclear morphology in EUE cell clones.Cell. Mol. Biol. 33, 73–81.
Ohta, N., Pardee, A. B., McAuslan, B. R. &Burger, M. M. (1968) Sialic acid content and controls of normal and malignant cells.Biochim. Biophys. Acta 158, 98–102.
Pearse, A. G. E. (1985) Carbohydrates and mucosubstances. InHistochemistry: Theoretical and Applied. Vol. 2, pp. 675–753. Edinburgh, London, New York: Churchill Livingstone.
Pellicciari, C., Garagna, S., Fuhrman Conti, A. M. &Manfredi Romanini, M. G. (1986) Effects of hypertonic medium on human cell growth-I. Feulgen-DNA content and chromosome number in EUE cells.Cell. Mol. Biol. 32, 173–81.
Rizzo, W. B. &Bustin, M. (1977) Lectins as probes of chromatin structure. Binding of Concanavalin A to purified rat liver chromatin.J. Biol. Chem. 252, 7062–7.
Robbins, F., Pederson, T. &Klein, P. (1970) Comparison of mitotic phenomena and effects induced by hypertonic solutions in HeLa cell.J. Cell Biol. 44, 400–16.
Roti Roti, W. L. &Rothstein, A. (1973a) Adaptation of mouse leukemic cells (L5178Y) to anisotonic media-I. Cell volume regulation.Exp. Cell Res. 79, 295–310.
Roti Roti, W. L. &Rothstein, A. (1973b) Adaptation of mouse leukemic cells (L5178Y) to anisotonic media-II. Cell growth.Exp. Cell Res. 79, 311–17.
Sevaljevic, L., Petrovic, S. L. &Petrovic, M. (1979) Binding ofLens culinaris lectin to sea urchin embryo chromatin.Experientia 35, 193–4.
Stein, G. S. &Kleinsmith, L. J. (eds) (1975)Chromosomal Proteins and their Role in the Regulation of Gene Expression, New York: Academic Press.
Stein, G. S., Roberts, R. M., Stein, J. L. &Davis, J. L. (1981) Nuclear glycoproteins and glycosaminoglycans. InThe Cell Nucleus, Vol. 9, pp. 341–57. New York: Academic Press.
Tenchini, M. L., Bolognani, L. &De Carli, L. (1980) Effect of hypertonicity on survival of unprotected human cultured cells following freezing and thawing.Cryobiology 17, 120–4.
Terni, M. &Lo Monaco, G. B. (1958) Coltura continua di cellule derivate da embrione umano.Lo Sperimentale 108, 177–85.
Zippel, R., Martegani, E., Vanoni, M., Mazzini, G. &Alberghina L. (1982) Cell cycle analysis in a human cell line (EUE cells).Cytometry 2, 426–30.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fantin, A.M.B., Menghi, G., Franchini, A. et al. Characterization of glycoconjugates in an embryonic human epithelial line and changes consequent to adaptation to a hyperosmotic medium. Histochem J 21, 79–88 (1989). https://doi.org/10.1007/BF01005983
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01005983