Summary
A polysaccharide surface coat was demonstrated in the various developmental stages of Leishmania donovani at the fine structural level by several cytochemical methods. During the course of in vitro cultivation, amastigote forms which had lost the outer membrane gave positive surface coat staining reactions following Alcian blue-lanthanum nitrate treatment. Coat matrix deposition appeared to increase during cultivation of the transforming amastigotes, reaching a maximum thickness at ∼7 hr characteristic of that observed with the promastigote stages. Differences were observed in the appearance of the surface coat materials in amastigotes treated with Alcian blue and lanthanum nitrate at 0°C and 24°C. Heavy deposits of coat-like material were observed in the flagellar tip region of promastigotes. Results of growth studies indicated that total rosette formation increased during the first 5 days of the promastigote growth phase. No differences were observed, however, in the appearance of the surface coat in promastigotes obtained from culture after various periods. Surface coat materials also were observed in promastigotes treated with purified ruthenium red and violet fractions. It was suggested that the surface coat might have a role in the adhesion of the organisms to various substrates and other cells in culture.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adler, A.: Leishmania. In: Advances in parasitology, vol. 2, (B. Dawes, ed.), p. 35–96. New York: Academic Press 1964
Allsopp, B. A., Njogu, A. R., Humphreys, K. C.: The nature and localization of Trypanosoma brucei subgroup exoantigen and its relationship to the 4S antigen. Exp. Parasit. 29, 271–284 (1971)
Behnke, O.: Electron microscopic observations on the surface coating of human blood platelets. J. Ultrastruct. Res. 24, 51–69 (1968)
Behnke, O., Zelander, T.: Preservation of intracellular substances by the cationic dye Alcian blue in preparative procedures for electron microscopy. J. Ultrastruct. Res. 31, 424–438 (1970)
Bennett, H. S.: Morphological aspects of extracellular polysaccharides. J. Histochem. Cytochem. 11, 14–23 (1963)
Brooker, B. E.: Flagellar attachment and detachment of Crithidia fasciculata to the gut wall of Anopheles gambiae. Protoplasma 73, 191–202 (1971)
Curtis, A. S. G.: Adhesive interactions between organisms. In: Functional aspects of parasite surfaces (A. E. R. Taylor and R. Muller, eds.), p. 1–21. London: Blackwell Scientific Publications 1972
D'Alesandro, P. A.: A serological comparison of the leishmaniform and the leptomonad stages of Leishmania donovani. M. A. Thesis, New Brunswick, N. J., Rutgers-The State University, p. 42 (1954)
Doganges, P. T., Schubert, M.: The use of lanthanum to study the degradation of a protein polysaccharide. J. biol. Chem. 239, 1498–1503 (1964)
Dwyer, D. M.: A monophasic medium for cultivating Leishmania donovani in large numbers. J. Parasit. 58, 847–848 (1972)
Dwyer, D. M.: Amastigote and promastigote stages of Leishmania donovani: an immunologic and immunochemical comparison. Proc. IVth Int. Congr. Protozool. 129 (1973)
Dwyer, D. M.: Lectin binding saccharides on a parasitic protozoan. Science, in press (1974)
Hughes, R. C.: Glycoproteins as components of cellular membranes. Progr. Biophys. molec. Biol. 26, 191–255 (1973)
Luft, J. H.: Improvement in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961)
Luft, J. H.: Ruthenium red and violet. I. Chemistry, purification, methods of use for electron microscopy and mechanism of action. Anat. Rec. 171, 347–368 (1971a)
Luft, J. H.: Ruthenium red and violet. II. Fine structural localization in animal tissues. Anat. Rec. 171, 369–416 (1971b)
Molyneux, D. H.: The fine-structure of the epimastigote forms of Trypanosoma lewisi in the rectum of the flea Nosopsyllus fasciatus. Parasitology 59, 55–66 (1969)
Njogu, A. R., Humphreys, K. C.: The nature of the 4S antigens of the brucei subgroup trypanosomes. Exp. Parasit. 31, 178–187 (1972)
Poste, G., Greenham, L. W., Mallucci, L., Reeve, P., Alexander, D. J.: The study of cellular “microexudates” by ellipsometry and their relationship to the cell coat. Exp. Cell Res. 78, 303–313 (1973)
Rambourg, A.: Morphological and histochemical aspects of glycoproteins at the surface of animal cells. Int. Rev. Cytol. 31, 57–114 (1971)
Rudzinska, M. A., D'Alesandro, P. A., Trager, W.: The fine structure of Leishmania donovani and the role of the kinetoplast in the leishmania-leptomonad transformation. J. Protozool. 11, 166–191 (1964)
Shea, S. M.: Lanthanum staining of the surface coat of cells, its enhancement by the use of fixatives containing Alcian blue or cetylpyridinium chloride. J. Cell Biol. 51, 611–620 (1971)
Simpson, L.: The leishmania-leptomonad transformation of Leishmania donovani: nutritional requirements, respiration changes and antigenic changes. J. Protozool. 15, 201–207 (1968)
Spiro, R. G.: Glycoproteins. Ann. Rev. Biochem. 39, 599–638 (1970)
Strauss, P.: The effect of homologous rabbit antiserum on the growth of Leishmania tarentolae—a fine structure study. J. Protozool. 18, 147–156 (1971)
Szirmai, J. A.: Quantitative approaches in the histochemistry of mucopolysaccharides. J. Histochem. Cytochem. 11, 24–34 (1963)
Szubinska, B., Luft, J. H.: Ruthenium red and violet. III. Fine structure of the plasma membrane and extraneous coats in amoebae (A. proteus and Chaos chaos). Anat. Rec. 171, 417–442 (1971)
Venable, J., Coggeshall, R.: The use of a simple lead citrate stain in electron microscopy. J. Cell Biol. 25, 407–408 (1965)
Vickerman, K.: Morphological and physiological considerations of extracellular blood protozoa. In: Ecology and physiology of parasites (A. M. Fallis, ed.), p. 58–91. Toronto: University of Toronto Press 1971
Vickerman, K.: The host-parasite interface of parasitic protozoa. Some problems posed by ultrastructural studies. In: Functional aspects or parasite surfaces (A. E. R. Taylor and R. Muller, eds.), p. 71–91. London: Blackwell Scientific Publications 1972
Vickerman, K.: The mode of attachment of Trypanosoma vivax in the proboscis of the tsetse fly Glossina fuscipes: an ultrastructural study of the epimastigote stage of the trypanosome. J. Protozool. 20, 394–404 (1973)
Wright, K. A., Hales, H.: Cytochemistry of the pellicle of bloodstream forms of Trypnaosoma (Trypanozoon) brucei. J. Parasit. 56, 671–683 (1970)
Author information
Authors and Affiliations
Additional information
Supported in part by Training Grant 5TOI-AI00192-12, 13 and Research Grant 5TOI-AIO8989-03, 04 from NIAID, U.S. Public Health Service.
Postdoctoral Research Fellow supported by Fellowship 5FO2-AI51048-01, 02 from NIAID, U.S. Public Health Service.
The authors wish to thank Prof. William Trager and Philip A. D'Alesandro of The Rockefeller University for their helpful comments and criticisms during the course of the present study and in the preparation of this manuscript. The excellent technical assistance of Dr. Earl Weidner during the initial phases of this work is gratefully acknowledged.
Rights and permissions
About this article
Cite this article
Dwyer, D.M., Langreth, S.G. & Dwyer, N.K. Evidence for a polysaccharide surface coat in the developmental stages of Leishmania donovani: A fine structure-cytochemical study. Z. Parasitenk. 43, 227–249 (1974). https://doi.org/10.1007/BF00328879
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00328879