[en] [en] BACKGROUND: In addition to its role in the digestive system, the peritrophic membrane (PM) provides a physical barrier protecting the intestine from abrasion and against pathogens. Because of its sensitivity to RNA interference (RNAi), the notorious pest insect, the Colorado potato beetle (CPB, Leptinotarsa decemlineata), has become a model insect for functional studies. Previously, RNAi-mediated silencing of Mannosidase-Ia (ManIa), a key enzyme in the transition from high-mannose glycan moieties to paucimannose N-glycans, was shown to disrupt the transition from larva to pupa and the metamorphosis into adult beetles. While these effects at the organismal level were interesting in a pest control context, the effects at the organ or tissue level and also immune effects have not been investigated yet. To fill this knowledge gap, we performed an analysis of the midgut and PM in ManIa-silenced insects.
RESULTS: As marked phenotype, the ManIaRNAi insects, the PM pore size was found to be decreased when compared to the control GFPRNAi insects. These smaller pores are related to the observation of thinner microvilli (Mv) on the epithelial cells of the midgut of ManIaRNAi insects. A midgut and PM proteome study and reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis with a selection of marker genes was performed to characterize the midgut cells and understand their response to the silencing of ManIa. In agreement with the loss of ManIa activity, an accumulation of high-mannose N-glycans was observed in the ManIa-silenced insects. As a pathogen-associated molecular pattern (PAMP), the presence of these glycan structures could trigger the activation of the immune pathways.
CONCLUSION: The observed decrease in PM pore size could be a response to prevent potential pathogens to access the midgut epithelium. This hypothesis is supported by the strong increase in transcription levels of the anti-fungal peptide drosomycin-like in ManIaRNAi insects, although further research is required to elucidate this possibility. The potential immune response in the midgut and the smaller pore size in the PM shed a light on the function of the PM as a physical barrier and provide evidence for the relation between the Mv and PM.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Liu, Dongdong; Laboratory of Agrozoology, Department Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
De Schutter, Kristof ; Laboratory of Agrozoology, Department Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
Far, Johann ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Staes, An; VIB Center for Medical Biotechnology, Ghent University, Ghent, Belgium ; Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
Dewettinck, Koen; Food Structure and Function Research Group, Department of Food Technology, Safety and Health, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
Quinton, Loïc ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie biologique
Gevaert, Kris; VIB Center for Medical Biotechnology, Ghent University, Ghent, Belgium ; Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
Smagghe, Guy ; Laboratory of Agrozoology, Department Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
Language :
English
Title :
RNAi of Mannosidase-Ia in the Colorado potato beetle and changes in the midgut and peritrophic membrane.
FWO - Fonds Wetenschappelijk Onderzoek Vlaanderen FEDER - Fonds Européen de Développement Régional UGent - Universiteit Gent ASCRS - American Society of Colon and Rectal Surgeons
Funding text :
Dongdong Liu is the recipient of a doctoral grant provided by China Scholarship Council. This research was supported by the Research Foundation-Flanders (FWO-Vlaanderen, Belgium) and the Special Research Fund from the Ghent University (Belgium). The authors also thank Bart De Pauw (Ghent University), Benny Lewille (Ghent University), Mathieu Tiquet (University of Liège), Nanou Tanteliarisoa (University of Liège) for their respective help with the critical point drying, SEM, and glycan sample preparation and MALDI mass spectra acquisition. The MALDI/ESI dual source FT-ICR solariX XR 9.4 T was co-funded by FEDER BIOMED HUB Technology Support (number 2.2.1/996).Dongdong Liu is the recipient of a doctoral grant provided by China Scholarship Council. This research was supported by the Research Foundation‐Flanders (FWO‐Vlaanderen, Belgium) and the Special Research Fund from the Ghent University (Belgium). The authors also thank Bart De Pauw (Ghent University), Benny Lewille (Ghent University), Mathieu Tiquet (University of Liège), Nanou Tanteliarisoa (University of Liège) for their respective help with the critical point drying, SEM, and glycan sample preparation and MALDI mass spectra acquisition. The MALDI/ESI dual source FT‐ICR solariX XR 9.4 T was co‐funded by FEDER BIOMED HUB Technology Support (number 2.2.1/996).
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