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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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