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Evidence for the Involvement of a Tsetse Midgut Lectin-Trypsin Complex in Differentiation of Bloodstream-Form Trypanosomes

Published online by Cambridge University Press:  19 September 2011

L. U. Abubakar ,
G. Zimba ,
C. Wells ,
F. Mulaa  and
E. O. Osir
L. U. Abubakar
Affiliation:
International Centre of Insect Physiology and Ecology, P. O. Box 30772, Nairobi, Kenya
G. Zimba
Affiliation:
University of Malawi, Bunda College of Agriculture, P. O. Box 219, Lilongwe, Malawi
C. Wells
Affiliation:
International Livestock Research Institute, P. O. Box 30709, Nairobi, Kenya
F. Mulaa
Affiliation:
Department of Biochemistry, University of Nairobi, P. O. Box 30197, Nairobi, Kenya
E. O. Osir*
Affiliation:
International Centre of Insect Physiology and Ecology, P. O. Box 30772, Nairobi, Kenya
*
E-mail: [email protected]
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Abstract

We have previously described a bloodmeal-induced molecule (lectin-trypsin complex) from the midgut of the tsetse fly, Glossina longipennis, with both lectin and trypsin activities (Osir et al., 1995). In this paper, we report on the isolation of a similar molecule from the midguts of Glossina fuscipes fnscipes and provide direct evidence for its involvement in the development of African trypanosomes. The molecule (native Mr ∼65,700) has two non-covalently linked subunits, Mr ∼28,800 and Mr ∼35,700. The native molecule was found to be capable of inducing differentiation of bloodstream-form trypanosomes into procyclic (midgut forms) in vitro. In the assays, specific antibodies against procyclin were used to monitor the transformation of the bloodstream-form trypanosomes into procyclic forms. This induction was specifically inhibited by D-glucosamine. Cis-aconitate was also capable of inducing the transformation process with the same efficiency as that of the lectin-trypsin complex. While increasing the concentrations of the lectin-trypsin complex (≥100 μg protein/ml) in the incubation assays resulted into higher transformation rates, it also led to high parasite mortality. These results provide evidence for the involvement of the midgut lectin-trypsin complex in the differentiation of bloodstream-form trypanosomes within tsetse midgut.

Résumé

Nous avons précédemment décrit une molécule induite par le repas de sang (un complexe lectine-trypsine) présente dans l'intestin moyen de la mouche tsé-tsé 'Glossina longipennis, ayant une activité lectine et trypsine (Osir et al., 1995). Dans ce papier, nous décrivons l'isolement d'une molécule similaire présente dans l'intestin moyen de Glossina fuscipes fuscines et démontrons son implication dans le développement des trypanosomes africains. La molécule (pure Mr ∼65,700) présente deux sous unités liées non covalentes, Mr ∼28,800 et Mr ∼35,700. La molécule pure est capable d'induire la différenciation des formes sanguines du trypanosome en formes pro-cycliques (intestin moyen) in vitro. Lors des essais, des anticorps spécifiques de la procycline ont été utilisés pour contrôler la transformation des formes sanguines du trypanosome en forme procycliques. Cette induction a été inhibée spécifiquement avec du D-glucosamine. Le cis-aconitate est également capable d'induire le processus de transformation avec une efficacité comparable à celle du complexe lectine-trypsine. Alors que l'augmentation des concentrations du complexe lectine-trypsine (≥100 μg protéine/ml) dans les essais d'incubation permet d'augmenter les taux de transformation, il induit également une importante mortalité du parasite. Ces résultats démontrent la participation du complexe lectine-trypsine de l'intestin moyen dans la différenciation des formes sanguines des trypanosomes dans l'intestin moyen de la mouche tsé-tsé.

Keywords

GlossinaTripanosoma bruceilectintrypsinGlossinaTripanosoma bruceilectinetrypsine
Type
Research Articles
Information
International Journal of Tropical Insect Science , Volume 23 , Issue 3 , September 2003 , pp. 197 - 205
Copyright
Copyright © ICIPE 2003

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