Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans
Tsetse flies are the principal insect vectors of African trypanosomes—sleeping sickness in humans and Nagana in cattle. One of the tsetse fly species, Glossina morsitans morsitans, is host to the parasite, Trypanosoma brucei, a major cause of African trypanosomiasis. Precise details of the life cycl...
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MDPI AG
2018-03-01
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| Series: | Pathogens |
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| Online Access: | http://www.mdpi.com/2076-0817/7/1/32 |
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| author | Evelyn Rogerson Julien Pelletier Alvaro Acosta-Serrano Clair Rose Sarah Taylor Scott Guimond Marcelo Lima Mark Skidmore Edwin Yates |
| author_facet | Evelyn Rogerson Julien Pelletier Alvaro Acosta-Serrano Clair Rose Sarah Taylor Scott Guimond Marcelo Lima Mark Skidmore Edwin Yates |
| author_sort | Evelyn Rogerson |
| collection | DOAJ |
| description | Tsetse flies are the principal insect vectors of African trypanosomes—sleeping sickness in humans and Nagana in cattle. One of the tsetse fly species, Glossina morsitans morsitans, is host to the parasite, Trypanosoma brucei, a major cause of African trypanosomiasis. Precise details of the life cycle have yet to be established, but the parasite life cycle involves crossing the insect peritrophic matrix (PM). The PM consists of the polysaccharide chitin, several hundred proteins, and both glycosamino- and galactosaminoglycan (GAG) polysaccharides. Owing to the technical challenges of detecting small amounts of GAG polysaccharides, their conclusive identification and composition have not been possible until now. Following removal of PMs from the insects and the application of heparinases (bacterial lyase enzymes that are specific for heparan sulphate (HS) GAG polysaccharides), dot blots with a HS-specific antibody showed heparan sulphate proteoglycans (HSPGs) to be present, consistent with Glossina morsitans morsitans genome analysis, as well as the likely expression of the HSPGs syndecan and perlecan. Exhaustive HS digestion with heparinases, fluorescent labeling of the resulting disaccharides with BODIPY fluorophore, and separation by strong anion exchange chromatography then demonstrated the presence of HS for the first time and provided the disaccharide composition. There were no significant differences in the type of disaccharide species present between genders or between ages (24 vs. 48 h post emergence), although the HS from female flies was more heavily sulphated overall. Significant differences, which may relate to differences in infection between genders or ages, were evident, however, in overall levels of 2-O-sulphation between sexes and, for females, between 24 and 48 h post-emergence, implying a change in expression or activity for the 2-O-sulphotransferase enzyme. The presence of significant quantities of disaccharides containing the monosaccharide GlcNAc6S contrasts with previous findings in Drosophila melanogaster and suggests subtle differences in HS fine structure between species of the Diptera. |
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| language | English |
| last_indexed | 2024-04-13T07:25:41Z |
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| spelling | doaj.art-4270412daf4b4a76aaad53967a6d2a412022-12-22T02:56:29ZengMDPI AGPathogens2076-08172018-03-01713210.3390/pathogens7010032pathogens7010032Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitansEvelyn Rogerson0Julien Pelletier1Alvaro Acosta-Serrano2Clair Rose3Sarah Taylor4Scott Guimond5Marcelo Lima6Mark Skidmore7Edwin Yates8Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UKMolecular & Structural Bioscience, School of Life Sciences, Keele University, Keele ST5 5BG, UKLiverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UKLiverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UKInstitute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB, UKInstitute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB, UKDepartment of Biochemistry, Universidade Federal de São Paulo, São Paulo 04044-020, BrazilMolecular & Structural Bioscience, School of Life Sciences, Keele University, Keele ST5 5BG, UKMolecular & Structural Bioscience, School of Life Sciences, Keele University, Keele ST5 5BG, UKTsetse flies are the principal insect vectors of African trypanosomes—sleeping sickness in humans and Nagana in cattle. One of the tsetse fly species, Glossina morsitans morsitans, is host to the parasite, Trypanosoma brucei, a major cause of African trypanosomiasis. Precise details of the life cycle have yet to be established, but the parasite life cycle involves crossing the insect peritrophic matrix (PM). The PM consists of the polysaccharide chitin, several hundred proteins, and both glycosamino- and galactosaminoglycan (GAG) polysaccharides. Owing to the technical challenges of detecting small amounts of GAG polysaccharides, their conclusive identification and composition have not been possible until now. Following removal of PMs from the insects and the application of heparinases (bacterial lyase enzymes that are specific for heparan sulphate (HS) GAG polysaccharides), dot blots with a HS-specific antibody showed heparan sulphate proteoglycans (HSPGs) to be present, consistent with Glossina morsitans morsitans genome analysis, as well as the likely expression of the HSPGs syndecan and perlecan. Exhaustive HS digestion with heparinases, fluorescent labeling of the resulting disaccharides with BODIPY fluorophore, and separation by strong anion exchange chromatography then demonstrated the presence of HS for the first time and provided the disaccharide composition. There were no significant differences in the type of disaccharide species present between genders or between ages (24 vs. 48 h post emergence), although the HS from female flies was more heavily sulphated overall. Significant differences, which may relate to differences in infection between genders or ages, were evident, however, in overall levels of 2-O-sulphation between sexes and, for females, between 24 and 48 h post-emergence, implying a change in expression or activity for the 2-O-sulphotransferase enzyme. The presence of significant quantities of disaccharides containing the monosaccharide GlcNAc6S contrasts with previous findings in Drosophila melanogaster and suggests subtle differences in HS fine structure between species of the Diptera.http://www.mdpi.com/2076-0817/7/1/32Glossina morsitans morsitanstsetse flyTrypanosoma bruceiheparan sulphateperitrophic matrix |
| spellingShingle | Evelyn Rogerson Julien Pelletier Alvaro Acosta-Serrano Clair Rose Sarah Taylor Scott Guimond Marcelo Lima Mark Skidmore Edwin Yates Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans Pathogens Glossina morsitans morsitans tsetse fly Trypanosoma brucei heparan sulphate peritrophic matrix |
| title | Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans |
| title_full | Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans |
| title_fullStr | Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans |
| title_full_unstemmed | Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans |
| title_short | Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans |
| title_sort | variations in the peritrophic matrix composition of heparan sulphate from the tsetse fly glossina morsitans morsitans |
| topic | Glossina morsitans morsitans tsetse fly Trypanosoma brucei heparan sulphate peritrophic matrix |
| url | http://www.mdpi.com/2076-0817/7/1/32 |
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