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Analysis of surface carbohydrates of Trichobilharzia ocellata miracidia and sporocysts using lectin binding techniques

Published online by Cambridge University Press:  06 April 2009

M. J. T. Gerhardus ,
J. M. C. Baggen ,
W. P. W. Van Der Knaap  and
T. Sminia
M. J. T. Gerhardus
Affiliation:
Department of Medical Microbiology and ParasitologyVrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
J. M. C. Baggen
Affiliation:
Department of Medical Microbiology and ParasitologyVrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
W. P. W. Van Der Knaap
Affiliation:
Department of Medical Microbiology and ParasitologyVrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
T. Sminia
Affiliation:
Department of Histology, Faculty of Medicine, Vrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
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Extract

Miracidia and in vitro-derived primary sporocysts of the avian schistosome Trichobilharzia ocellata were studied for the expression and the characteristics of glycoconjugate moieties comprising the surface coat. Using a panel of 9 peroxidase labelled lectins, several different lectin binding sites were demonstrated on the larvae. Fixed miracidia have binding sites for 7 of the lectins; wheat-germ agglutinin binds to both the ciliated plates and the tegumental ridges between them; the other 6 lectins bind to the plates only. Three of the miracidia-binding lectins, wheat-germ agglutinin, concanavalin A and peanut agglutinin, also bind to fixed sporocysts. Since the miracidial ridges are devoid of binding sites for concanavalin A and peanut agglutinin, whereas the sporocyst tegument binds these lectins, it appears that these sites become exposed during or shortly after transformation. In saturation experiments, low concentrations of peanut agglutinin and concanavalin A are bound more avidly by sporocysts than by miracidia, indicating a higher binding affinity of the former. The two larval forms do not differ in affinity for wheat-germ agglutinin but they have different binding capacities; when offered in high concentrations, more of this lectin is bound by sporocysts than by miracidia. Lectin binding was competitively inhibited by adding the appropriate free saccharides. Live larvae showed the same lectin binding pattern as did fixed specimens. Proteinase treatment reduced lectin binding to living and, to a lesser extent, to fixed larvae, suggesting that binding sites are constituents of proteoglycoconjugates. After SDS–PAGE of extracts from miracidia and sporocysts and subsequent Western blotting, some of the lectins failed to bind glycoproteins, others reacted with an array of bands. The patterns differed among the lectins and each lectin gave different patterns for miracidia and sporocysts. The results obtained with these two lectin-binding techniques support the conclusion that stage-specific proteoglycoconjugates occur at the surface of T. ocellata larvae.

Keywords

Trichobilharzia ocellataschistosomeglycoproteinsligand blottinglectins
Type
Research Article
Information
Parasitology , Volume 103 , Issue 1 , August 1991 , pp. 51 - 59
Copyright
Copyright © Cambridge University Press 1991

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References

Bayne, C. J., Loker, E. S. & Yui, M. A. (1986). Interactions between the plasma proteins of Biomphalaria glabrata (Gastropoda) and the sporocyst tegument of Schistosoma mansoni (Trematoda). Parasitology 92, 653–64.CrossRefGoogle ScholarPubMed
Boswell, C. A. & Bayne, C. J. (1986). Lectin-dependent cell-mediated cytotoxicity in an invertebrate model: Con A does not act as a bridge. Immunology 57, 261–4.Google Scholar
Boswell, C. A., Yoshino, T. P. & Dunn, T. S. (1987). Analysis of tegumental surface proteins of Schistosoma mansoni primary sporocysts. Journal of Parasitology 73, 778–86.CrossRefGoogle ScholarPubMed
Dunn, T. S. & Yoshino, T. P. (1988). Schistosoma mansoni: origin and expression of a tegumental surface antigen on the miracidium and primary sporocyst. Experimental Parasitology 67, 167–81.CrossRefGoogle ScholarPubMed
Hayunga, E. G. & Sumner, M. P. (1986). Characterization of surface glycoproteins on Schistosoma mansoni adult worms by lectin affinity chromatography. Journal of Parasitology 72, 283–91.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Macgregor, A. N., Stott, D. I. & Kusel, J. R. (1985). Lectin binding to glycoproteins in the surface membrane of Schistosoma mansoni. Molecular and Biochemical Parasitology 16, 163–72.CrossRefGoogle ScholarPubMed
Mellink, J. J. & Van Den Bovenkamp, W. (1985). In vitro culture of intramolluscan stages of the avian schistosome Trichobilharzia ocellata. Zeitschrift für Parasitenkunde 71, 337–51.CrossRefGoogle ScholarPubMed
Meuleman, E. A., Huyer, A. R. & Mooij, J. H. (1984). Maintenance of the life cycle of Trichobilharzia ocellata via the duck Anas platyrhynchos and the pond snail Lymnaea stagnalis. Netherlands Journal of Zoology 34, 414–17.CrossRefGoogle Scholar
Meuleman, E. A., Lyaruu, D. M., Khan, M. A., Holzmann, P. J. & Sminia, T. (1978). Ultrastructural changes in the body wall of Schistosoma mansoni during the transformation of the miracidium into the mother sporocyst in the snail host Biomphalaria pfeifferi. Zeitschrift für Parasitenkunde 56, 227–42.CrossRefGoogle ScholarPubMed
Renwrantz, L. (1986). Lectins in molluscs and arthropods: their occurrence, origin and roles in immunity. In Immune Mechanisms in Invertebrate Vectors (ed. Lackie, A. M.), pp. 81–93. Oxford: Clarendon Press.Google Scholar
Schallig, H. D. F. H., Schut, A., Van Der Knaap, W. P. W. & De Jong-Brink, M. (1990). A simplified medium for the in vitro culture of mother sporocysts of the schistosome Trichobilharzia ocellata. Parasitology Research 76, 278–9.CrossRefGoogle Scholar
Simpson, A. J. G. (1990). Schistosome surface antigens: developmental expression and immunological function. Parasitology Today 6, 40–5.CrossRefGoogle ScholarPubMed
Simpson, A. J. G., Correa-Oliveira, R., Smithers, S. R. & Sher, A. (1983). The exposed carbohydrates of schistosomula of Schistosoma mansoni and their modification during maturation in vivo. Molecular and Biochemical Parasitology 8, 191–205.CrossRefGoogle ScholarPubMed
Simpson, A. J. G. & Smithers, S. R. (1980). Characterization of the exposed carbohydrates on the surface membranes of adult Schistosoma mansoni by analysis of lectin binding. Parasitology 81, 1–15.CrossRefGoogle ScholarPubMed
Sokal, R. R. & Rohlf, F. J. (1969). Biometry. The Principles and Practice of Statistics in Biological Research. San Francisco: W. H. Freeman.Google Scholar
Towbin, H., Staehelin, T. & Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350–4.CrossRefGoogle ScholarPubMed
Van Der Knaap, W. P. W. & Loker, E. S. (1990). Immune mechanisms in trematode–snail interactions. Parasitology Today 6, 175–82.CrossRefGoogle ScholarPubMed
Yoshino, T. P., Cheng, T. C. & Renwrantz, L. R. (1977). Lectin and human blood group determinants of Schistosoma mansoni: alterations following in vitro transformation of miracidium to mother sporocyst. Journal of Parasitology 63, 818–24.CrossRefGoogle ScholarPubMed
Zelck, U. & Becker, W. (1990). Lectin binding to cells of Schistosoma mansoni sporocysts and surrounding Biomphalaria glabrata tissue. Journal of Invertebrate Pathology 55, 93–9.CrossRefGoogle ScholarPubMed