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Alvaro Acosta-serrano - One of the best experts on this subject based on the ideXlab platform.
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Trypanosoma brucei colonises the tsetse gut via an immature Peritrophic Matrix in the proventriculus
2019Co-Authors: Clair Rose, Naomi A Dyer, Carla Solorzano, Alison J Beckett, Ben Middlehurst, Marco Marcello, Michael J. Lehane, Aitor Casas-sanchez, Ian A. Prior, Alvaro Acosta-serranoAbstract:Abstract The Peritrophic Matrix (PM) of haematophagus insects is a chitinous structure that surrounds the bloodmeal, forming a protective barrier against oral pathogens and abrasive particles. To establish an infection in the tsetse midgut, Trypanosoma brucei must colonise the ectoPeritrophic space (ES), located between the PM and gut epithelium. Although unproven, it is generally accepted that trypanosomes reach the ES by directly penetrating the PM in the anterior midgut. Here we revisited this event by employing novel fluorescence and electron microscopy methodologies and found that instead, trypanosomes reach the ES via the newly secreted PM in the tsetse proventriculus. Within this model, parasites colonising the proventriculus can either migrate to the ES or become trapped within PM layers forming cysts that move along the entire gut as the PM gets remodelled. Early proventricular colonisation appears to be promoted by unidentified factors in trypanosome-infected blood, resulting in higher salivary gland infections and potentially increasing parasite transmission.
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An Investigation into the Protein Composition of the Teneral Glossina morsitans morsitans Peritrophic Matrix
2016Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Gemma Molyneux, Jonathan Wastling, Alvaro Acosta-serranoAbstract:Background: Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism. Methods: PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS. Results: Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM
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An Investigation into the Protein Composition of the Teneral Glossina morsitans morsitans Peritrophic Matrix
PLoS neglected tropical diseases, 2014Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Jonathan Wastling, Gemma R. Molyneux, Alvaro Acosta-serranoAbstract:BACKGROUND Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism. METHODS PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS. RESULTS Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM. CONCLUSION To our knowledge this is the first report on the protein composition of teneral G. m. morsitans, an important vector of African trypanosomes. Further functional analyses of these proteins will lead to a better understanding of the tsetse physiology and may help identify potential molecular targets to block trypanosome development within the tsetse.
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An investigation into the protein composition of the teneral Glossina morsitans morsitans Peritrophic Matrix.
Public Library of Science (PLoS), 2014Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Gemma Molyneux, Jonathan Wastling, Alvaro Acosta-serranoAbstract:Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism.PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS.Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM.To our knowledge this is the first report on the protein composition of teneral G. m. morsitans, an important vector of African trypanosomes. Further functional analyses of these proteins will lead to a better understanding of the tsetse physiology and may help identify potential molecular targets to block trypanosome development within the tsetse
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Categorization of the G. m. morsitans Peritrophic Matrix proteins as identified through LC-MS/MS according to their putative functions.
2014Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Gemma Molyneux, Jonathan Wastling, Michael J. Lehane, Alvaro Acosta-serranoAbstract:Categorization of the G. m. morsitans Peritrophic Matrix proteins as identified through LC-MS/MS according to their putative functions.
Clair Rose - One of the best experts on this subject based on the ideXlab platform.
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trypanosoma brucei colonizes the tsetse gut via an immature Peritrophic Matrix in the proventriculus
Nature microbiology, 2020Co-Authors: Clair Rose, Lee R. Haines, Aitor Casassanchez, Naomi A Dyer, Carla Solorzano, Alison J Beckett, Ben Middlehurst, Marco Marcello, Jaime Lisack, Markus EngstlerAbstract:The Peritrophic Matrix of blood-feeding insects is a chitinous structure that forms a protective barrier against oral pathogens and abrasive particles1. Tsetse flies transmit Trypanosoma brucei, which is the parasite that causes human sleeping sickness and is also partially responsible for animal trypanosomiasis in Sub-Saharan Africa. For this parasite to establish an infection in flies, it must first colonize the area between the Peritrophic Matrix and gut epithelium called the ectoPeritrophic space. Although unproven, it is generally accepted that trypanosomes reach the ectoPeritrophic space by penetrating the Peritrophic Matrix in the anterior midgut2-4. Here, we revisited this event using fluorescence- and electron-microscopy methodologies. We show that trypanosomes penetrate the ectoPeritrophic space in which the newly made Peritrophic Matrix is synthesized by the proventriculus. Our model describes how these proventriculus-colonizing parasites can either migrate to the ectoPeritrophic space or become trapped within Peritrophic Matrix layers to form cyst-like bodies that are passively pushed along the gut as the Matrix gets remodelled. Furthermore, early proventricular colonization seems to be promoted by factors in trypanosome-infected blood that cause higher salivary gland infections and potentially increase parasite transmission.
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Trypanosoma brucei colonises the tsetse gut via an immature Peritrophic Matrix in the proventriculus
2019Co-Authors: Clair Rose, Naomi A Dyer, Carla Solorzano, Alison J Beckett, Ben Middlehurst, Marco Marcello, Michael J. Lehane, Aitor Casas-sanchez, Ian A. Prior, Alvaro Acosta-serranoAbstract:Abstract The Peritrophic Matrix (PM) of haematophagus insects is a chitinous structure that surrounds the bloodmeal, forming a protective barrier against oral pathogens and abrasive particles. To establish an infection in the tsetse midgut, Trypanosoma brucei must colonise the ectoPeritrophic space (ES), located between the PM and gut epithelium. Although unproven, it is generally accepted that trypanosomes reach the ES by directly penetrating the PM in the anterior midgut. Here we revisited this event by employing novel fluorescence and electron microscopy methodologies and found that instead, trypanosomes reach the ES via the newly secreted PM in the tsetse proventriculus. Within this model, parasites colonising the proventriculus can either migrate to the ES or become trapped within PM layers forming cysts that move along the entire gut as the PM gets remodelled. Early proventricular colonisation appears to be promoted by unidentified factors in trypanosome-infected blood, resulting in higher salivary gland infections and potentially increasing parasite transmission.
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An Investigation into the Protein Composition of the Teneral Glossina morsitans morsitans Peritrophic Matrix
2016Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Gemma Molyneux, Jonathan Wastling, Alvaro Acosta-serranoAbstract:Background: Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism. Methods: PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS. Results: Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM
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An Investigation into the Protein Composition of the Teneral Glossina morsitans morsitans Peritrophic Matrix
PLoS neglected tropical diseases, 2014Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Jonathan Wastling, Gemma R. Molyneux, Alvaro Acosta-serranoAbstract:BACKGROUND Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism. METHODS PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS. RESULTS Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM. CONCLUSION To our knowledge this is the first report on the protein composition of teneral G. m. morsitans, an important vector of African trypanosomes. Further functional analyses of these proteins will lead to a better understanding of the tsetse physiology and may help identify potential molecular targets to block trypanosome development within the tsetse.
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An investigation into the protein composition of the teneral Glossina morsitans morsitans Peritrophic Matrix.
Public Library of Science (PLoS), 2014Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Gemma Molyneux, Jonathan Wastling, Alvaro Acosta-serranoAbstract:Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism.PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS.Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM.To our knowledge this is the first report on the protein composition of teneral G. m. morsitans, an important vector of African trypanosomes. Further functional analyses of these proteins will lead to a better understanding of the tsetse physiology and may help identify potential molecular targets to block trypanosome development within the tsetse
Kent S. Shelby - One of the best experts on this subject based on the ideXlab platform.
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detection of alternative splicing in western corn rootworm diabrotica virgifera virgifera leconte in association with ecry3 1ab resistance using rna seq and pacbio iso seq
Insect Molecular Biology, 2021Co-Authors: Z. Zhao, Bruce E Hibbard, Christine G. Elsik, Kent S. ShelbyAbstract:Alternative splicing is a common feature in eukaryotes that not only increases the transcript diversity, but also has have functional consequences. In insects, alternative splicing has been found associated with resistance to pesticides and Bt toxins. Up to date the alternative splicing in western corn rootworm (Diabrotica virgifera virgifera LeConte) has not been studied. To investigate its alternative splicing pattern and relation to Bt resistance, we carried out single-molecule real-time (SMRT) transcript sequencing and Iso-seq analysis on resistant, eCry3.1Ab-selected and susceptible, unselected, western corn rootworm neonate midguts which fed on seedling maize with and without eCry3.1Ab for 12 and 24 hours. We present transcriptome-wide alternative splicing patterns of western corn rootworm midgut in response to feeding on eCry3.1Ab-expressing corn using a comprehensive approach that combines both RNA-seq and SMRT transcript sequencing techniques. The results showed genes in western corn rootworm are highly alternatively spliced, which happens on 67.73% of multi-exon genes. One of the alternative splicing events we identified was a novel Peritrophic Matrix protein with two alternative splicing isoforms. Analysis of differential exon usage between resistant and susceptible colonies showed that in eCry3.1Ab-resistant western corn rootworm, expression of one isoform was significantly higher than in the susceptible colony, while no significant differences between colonies were observed with the other isoform. Our results provide the first survey of alternative splicing in western corn rootworm and suggest that the observed alternatively spliced isoforms of Peritrophic Matrix protein may be associated with eCry3.1Ab resistance in western corn rootworm.
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Detection of alternative splicing in western corn rootworm (Diabrotica virgifera virgifera LeConte) in association with Bt resistance using RNA-seq and PacBio Iso-Seq
2020Co-Authors: Z. Zhao, Bruce E Hibbard, Christine G. Elsik, Kent S. ShelbyAbstract:BackgroundAlternative splicing is one of the major mechanisms that increases transcriptome diversity in eukaryotes, including insect species that have gained resistance to pesticides and Bt toxins. In western corn rootworm (Diabrotica virgifera virgifera LeConte), neither alternative splicing nor its role in resistance to Bt toxins has been studied. ResultsTo investigate the mechanisms of Bt resistance we carried out single-molecule real-time (SMRT) transcript sequencing and Iso-seq analysis on resistant, eCry3.1Ab-selected and susceptible, unselected, western corn rootworm neonate midguts which fed on seedling maize with and without eCry3.1Ab for 12 and 24 hours. We present transcriptome-wide alternative splicing patterns of western corn rootworm midgut in response to feeding on eCry3.1Ab-expressing corn using a comprehensive approach that combines both RNA-seq and SMRT transcript sequencing techniques. We found that 67.73% of multi-exon genes are alternatively spliced, which is consistent with the high transposable element content of the genome. One of the alternative splicing events we identified was a novel Peritrophic Matrix protein with two alternative splicing isoforms. Analysis of differential exon usage between resistant and susceptible colonies showed that in eCry3.1Ab-resistant western corn rootworm, expression of one isoform was significantly higher than in the susceptible colony, while no significant differences between colonies were observed with the other isoform. ConclusionOur results provide the first survey of alternative splicing in western corn rootworm and suggest that the observed alternatively spliced isoforms of Peritrophic Matrix protein may be associated with eCry3.1Ab resistance in western corn rootworm.
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detection of alternative splicing in diabrotica virgifera virgifera leconte in association with bt resistance using rna seq and pacbio iso seq
bioRxiv, 2020Co-Authors: Z. Zhao, Bruce E Hibbard, Kent S. Shelby, Christine G. ElsikAbstract:Abstract Background Alternative splicing is one of the major mechanisms that increases transcriptome diversity in eukaryotes, including insect species that have gained resistance to pesticides and Bt toxins. In western corn rootworm (Diabrotica virgifera virgifera LeConte), neither alternative splicing nor its role in resistance to Bt toxins has been studied. Results To investigate the mechanisms of Bt resistance we carried out single-molecule real-time (SMRT) transcript sequencing and Iso-seq analysis on resistant, eCry3.1Ab-selected and susceptible, unselected, western corn rootworm neonate midguts which fed on seedling maize with and without eCry3.1Ab for 12 and 24 hours. We present transcriptome-wide alternative splicing patterns of western corn rootworm midgut in response to feeding on eCry3.1Ab-expressing corn using a comprehensive approach that combines both RNA-seq and SMRT transcript sequencing techniques. We found that 67.73% of multi-exon genes are alternatively spliced, which is consistent with the high transposable element content of the genome. One of the alternative splicing events we identified was a novel Peritrophic Matrix protein with two alternative splicing isoforms. Analysis of differential exon usage between resistant and susceptible colonies showed that in eCry3.1Ab-resistant western corn rootworm, expression of one isoform was significantly higher than in the susceptible colony, while no significant differences between colonies were observed with the other isoform. Conclusion Our results provide the first survey of alternative splicing in western corn rootworm and suggest that the observed alternatively spliced isoforms of Peritrophic Matrix protein may be associated with eCry3.1Ab resistance in western corn rootworm.
Dwayne D. Hegedus - One of the best experts on this subject based on the ideXlab platform.
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Identification of the Mamestra configurata (Lepidoptera: Noctuidae) Peritrophic Matrix proteins and enzymes involved in Peritrophic Matrix chitin metabolism.
Insect Science, 2015Co-Authors: Umut Toprak, Martin A. Erlandson, Cedric Gillott, Doug Baldwin, Cathy Coutu, Steve Karcz, Lianglu Wan, Dwayne D. HegedusAbstract:The Peritrophic Matrix (PM) is essential for insect digestive system physiology as it protects the midgut epithelium from damage by food particles, pathogens, and toxins. The PM is also an attractive target for development of new pest control strategies due to its per os accessibility. To understand how the PM performs these functions, the molecular architecture of the PM was examined using genomic and proteomic approaches in Mamestra configurata (Lepidoptera: Noctuidae), a major pest of cruciferous oilseed crops in North America. Liquid chromatography-tandem mass spectrometry analyses of the PM identified 82 proteins classified as: (i) peritrophins, including a new class with a CBDIII domain; (ii) enzymes involved in chitin modification (chitin deacetylases), digestion (serine proteases, aminopeptidases, carboxypeptidases, lipases and α-amylase) or other reactions (β-1,3-glucanase, alkaline phosphatase, dsRNase, astacin, pantetheinase); (iii) a heterogenous group consisting of polycalin, REPATs, serpin, C-Type lectin and Lsti99/Lsti201 and 3 novel proteins without known orthologs. The genes encoding PM proteins were expressed predominantly in the midgut. cDNAs encoding chitin synthase-2 (McCHS-2), chitinase (McCHI), and β-N-acetylglucosaminidase (McNAG) enzymes, involved in PM chitin metabolism, were also identified. McCHS-2 expression was specific to the midgut indicating that it is responsible for chitin synthesis in the PM, the only chitinous material in the midgut. In contrast, the genes encoding the chitinolytic enzymes were expressed in multiple tissues. McCHS-2, McCHI, and McNAG were expressed in the midgut of feeding larvae, and NAG activity was present in the PM. This information was used to generate an updated model of the lepidopteran PM architecture.
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Spatial and temporal synthesis of Mamestra configurata Peritrophic Matrix through a larval stadium.
Insect Biochemistry and Molecular Biology, 2014Co-Authors: Umut Toprak, Doug Baldwin, Cathy Coutu, Dwayne D. Hegedus, Martin A. ErlandsonAbstract:The structure and synthesis of the Mamestra configurata Peritrophic Matrix (PM) was examined at various time points during a larval stadium. Bright field and confocal fluorescence microscopy revealed major differences between the PM of feeding and molting larvae. The PM from feeding larvae was thinner and composed of approximately 5-10 layers. In contrast, mid-molt larvae had a chitinaceaous PM composed of multiple thick layers which filled most of the midgut lumen. PM synthesis initiates in the anterior midgut, based on the expression of genes encoding chitin synthase-2 (CHS-2), coincident with the incorporation of the major structural PM proteins (McIIM1, McIIM2 and McPM1). This is followed by reinforcement with other PM proteins (McIIM3 and McIIM4) as it moves toward the posterior of the midgut. Chitin deacetylase (McCDA1) was associated only with the anterior PM. Collectively, these findings indicate that the structural properties of the PM differ along the length of the midgut. Genes encoding chitinolytic enzymes (McCHI and McNAG) were expressed and exochitinase activity was present when the PM had degraded (pre-molt) and when the new PM was forming (mid-molt), indicating that they are involved in either PM turnover and/or maintenance dependent upon the stage.
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Role of enhancin in Mamestra configurata nucleopolyhedrovirus virulence: selective degradation of host Peritrophic Matrix proteins.
Journal of General Virology, 2012Co-Authors: Umut Toprak, Cedric Gillott, David A. Theilmann, Dwayne D. Hegedus, Douglas Baldwin, Stephanie Harris, Martin A. ErlandsonAbstract:To infect per os, baculovirus virions cross the Peritrophic Matrix (PM) to reach the midgut epithelium. Insect intestinal mucins (IIMs) are PM proteins that protect the PM and aid passage of the food bolus through the gut. Some baculoviruses, including Mamestra configurata nucleopolyhedrovirus (MacoNPV-A), encode metalloproteases, known as enhancins, that facilitate infection by degrading IIMs. We examined the interaction between MacoNPV-A enhancin and M. configurata IIMs both in vivo and in vitro. Per os inoculation of M. configurata larvae with MacoNPV-A occlusion bodies (OBs) resulted in the degradation of McIIM4 within 4 h of OB ingestion, while McIIM2 was unaffected. The PM recovered by 8 h post-inoculation. To investigate whether enhancin was responsible for the degradation of IIM, a recombinant Autographa californica multiple nucleopolyhedrovirus expressing MacoNPV enhancin (AcMNPV-enMP2) was constructed. Enhancin was found to be a component of occlusion-derived virions in AcMNPV-enMP2 and MacoNPV-A. In in vitro assays, McIIM4 was degraded after MacoNPV-A and AcMNPV-enMP2 treatments. Degradation of McIIM4 was inhibited by EDTA, a metalloprotease inhibitor, indicating that the degradation was due to enhancin activity. Thus, MacoNPV-A enhancin is able to degrade major structural PM proteins, but exhibits target substrate specificity.
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Insect intestinal mucins and serine proteases associated with the Peritrophic Matrix from feeding, starved and moulting Mamestra configurata larvae.
Insect Molecular Biology, 2010Co-Authors: Umut Toprak, Martin A. Erlandson, Cedric Gillott, Doug Baldwin, Dwayne D. HegedusAbstract:Insect intestinal mucins (McIIM2-4) expressed in the midgut of feeding, starved and moulting Mamestra configurata larvae were identified. McIIM2 and McIIM4 were associated with the Peritrophic Matrix (PM). PMs from feeding and starved larvae were translucent and contained organized chitin bundles perpendicular to their long axis, whereas PM from moulting larvae consisted of an inner opaque mass surrounded by an outer translucent sleeve. Serine protease genes (McSP1, McSP2, McSP25 and McSP29) were also expressed in these larvae and several serine proteases were associated with the PM. Serine protease activity was also detected in the midgut of feeding, starved and moulting larvae.
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new insights into Peritrophic Matrix synthesis architecture and function
Annual Review of Entomology, 2009Co-Authors: Dwayne D. Hegedus, Martin A. Erlandson, Cedric Gillott, Umut ToprakAbstract:The Peritrophic Matrix (PM) is a chitin and glycoprotein layer that lines the invertebrate midgut. Although structurally different, it is functionally similar to the mucous secretions of the vertebrate digestive tract. The PM is a physical barrier, protecting the midgut epithelium from abrasive food particles, digestive enzymes, and pathogens infectious per os. It is also a biochemical barrier, sequestering and, in some cases, inactivating ingested toxins. Finally, the PM compartmentalizes digestive processes, allowing for efficient nutrient acquisition and reuse of hydrolytic enzymes. The PM consists of an organized lattice of chitin fibrils held together by chitin binding proteins. Glycans fill the interstitial spaces, creating a molecular sieve, the properties of which are dependent on the immediate ion content and pH. In this review, we have integrated recent structural and functional information to create a holistic model for the PM. We also show how this information may generate novel technologies for use in insect pest management.
Lee R. Haines - One of the best experts on this subject based on the ideXlab platform.
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trypanosoma brucei colonizes the tsetse gut via an immature Peritrophic Matrix in the proventriculus
Nature microbiology, 2020Co-Authors: Clair Rose, Lee R. Haines, Aitor Casassanchez, Naomi A Dyer, Carla Solorzano, Alison J Beckett, Ben Middlehurst, Marco Marcello, Jaime Lisack, Markus EngstlerAbstract:The Peritrophic Matrix of blood-feeding insects is a chitinous structure that forms a protective barrier against oral pathogens and abrasive particles1. Tsetse flies transmit Trypanosoma brucei, which is the parasite that causes human sleeping sickness and is also partially responsible for animal trypanosomiasis in Sub-Saharan Africa. For this parasite to establish an infection in flies, it must first colonize the area between the Peritrophic Matrix and gut epithelium called the ectoPeritrophic space. Although unproven, it is generally accepted that trypanosomes reach the ectoPeritrophic space by penetrating the Peritrophic Matrix in the anterior midgut2-4. Here, we revisited this event using fluorescence- and electron-microscopy methodologies. We show that trypanosomes penetrate the ectoPeritrophic space in which the newly made Peritrophic Matrix is synthesized by the proventriculus. Our model describes how these proventriculus-colonizing parasites can either migrate to the ectoPeritrophic space or become trapped within Peritrophic Matrix layers to form cyst-like bodies that are passively pushed along the gut as the Matrix gets remodelled. Furthermore, early proventricular colonization seems to be promoted by factors in trypanosome-infected blood that cause higher salivary gland infections and potentially increase parasite transmission.
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An Investigation into the Protein Composition of the Teneral Glossina morsitans morsitans Peritrophic Matrix
2016Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Gemma Molyneux, Jonathan Wastling, Alvaro Acosta-serranoAbstract:Background: Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism. Methods: PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS. Results: Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM
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An Investigation into the Protein Composition of the Teneral Glossina morsitans morsitans Peritrophic Matrix
PLoS neglected tropical diseases, 2014Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Jonathan Wastling, Gemma R. Molyneux, Alvaro Acosta-serranoAbstract:BACKGROUND Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism. METHODS PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS. RESULTS Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM. CONCLUSION To our knowledge this is the first report on the protein composition of teneral G. m. morsitans, an important vector of African trypanosomes. Further functional analyses of these proteins will lead to a better understanding of the tsetse physiology and may help identify potential molecular targets to block trypanosome development within the tsetse.
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An investigation into the protein composition of the teneral Glossina morsitans morsitans Peritrophic Matrix.
Public Library of Science (PLoS), 2014Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Gemma Molyneux, Jonathan Wastling, Alvaro Acosta-serranoAbstract:Tsetse flies serve as biological vectors for several species of African trypanosomes. In order to survive, proliferate and establish a midgut infection, trypanosomes must cross the tsetse fly Peritrophic Matrix (PM), which is an acellular gut lining surrounding the blood meal. Crossing of this multi-layered structure occurs at least twice during parasite migration and development, but the mechanism of how trypanosomes do so is not understood. In order to better comprehend the molecular events surrounding trypanosome penetration of the tsetse PM, a mass spectrometry-based approach was applied to investigate the PM protein composition using Glossina morsitans morsitans as a model organism.PMs from male teneral (young, unfed) flies were dissected, solubilised in urea/SDS buffer and the proteins precipitated with cold acetone/TCA. The PM proteins were either subjected to an in-solution tryptic digestion or fractionated on 1D SDS-PAGE, and the resulting bands digested using trypsin. The tryptic fragments from both preparations were purified and analysed by LC-MS/MS.Overall, nearly 300 proteins were identified from both analyses, several of those containing signature Chitin Binding Domains (CBD), including novel peritrophins and peritrophin-like glycoproteins, which are essential in maintaining PM architecture and may act as trypanosome adhesins. Furthermore, 27 proteins from the tsetse secondary endosymbiont, Sodalis glossinidius, were also identified, suggesting this bacterium is probably in close association with the tsetse PM.To our knowledge this is the first report on the protein composition of teneral G. m. morsitans, an important vector of African trypanosomes. Further functional analyses of these proteins will lead to a better understanding of the tsetse physiology and may help identify potential molecular targets to block trypanosome development within the tsetse
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Categorization of the G. m. morsitans Peritrophic Matrix proteins as identified through LC-MS/MS according to their putative functions.
2014Co-Authors: Clair Rose, Lee R. Haines, Rodrigo Belmonte, Stuart D. Armstrong, Gemma Molyneux, Jonathan Wastling, Michael J. Lehane, Alvaro Acosta-serranoAbstract:Categorization of the G. m. morsitans Peritrophic Matrix proteins as identified through LC-MS/MS according to their putative functions.