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Because of its sensitivity to RNA interference (RNAi), the notorious pest insect, the Colorado potato beetle (CPB, <jats:italic>Leptinotarsa decemlineata<\/jats:italic>), has become a model insect for functional studies. Previously, RNAi\u2010mediated silencing of <jats:italic>Mannosidase\u2010Ia<\/jats:italic> (<jats:italic>ManIa<\/jats:italic>), a key enzyme in the transition from high\u2010mannose glycan moieties to paucimannose <jats:italic>N<\/jats:italic>\u2010glycans, 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 <jats:italic>ManIa<\/jats:italic>\u2010silenced insects.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>As marked phenotype, the <jats:italic>ManIa<\/jats:italic><jats:sup><jats:italic>RNAi<\/jats:italic><\/jats:sup> insects, the PM pore size was found to be decreased when compared to the control <jats:italic>GFP<\/jats:italic><jats:sup><jats:italic>RNAi<\/jats:italic><\/jats:sup> insects. These smaller pores are related to the observation of thinner microvilli (Mv) on the epithelial cells of the midgut of <jats:italic>ManIa<\/jats:italic><jats:sup><jats:italic>RNAi<\/jats:italic><\/jats:sup> insects. A midgut and PM proteome study and reverse transcription quantitative polymerase chain reaction (RT\u2010qPCR) analysis with a selection of marker genes was performed to characterize the midgut cells and understand their response to the silencing of <jats:italic>ManIa<\/jats:italic>. In agreement with the loss of <jats:italic>ManIa<\/jats:italic> activity, an accumulation of high\u2010mannose <jats:italic>N<\/jats:italic>\u2010glycans was observed in the <jats:italic>ManIa<\/jats:italic>\u2010silenced insects. As a pathogen\u2010associated molecular pattern (PAMP), the presence of these glycan structures could trigger the activation of the immune pathways.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>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\u2010fungal peptide <jats:italic>drosomycin\u2010like<\/jats:italic> in <jats:italic>ManIa<\/jats:italic><jats:sup><jats:italic>RNAi<\/jats:italic><\/jats:sup> 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. \u00a9 2022 Society of Chemical Industry.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ps.7145","type":"journal-article","created":{"date-parts":[[2022,8,23]],"date-time":"2022-08-23T11:38:03Z","timestamp":1661254683000},"page":"5071-5079","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["<scp>RNAi<\/scp> of <scp><i>Mannosidase\u2010Ia<\/i><\/scp> in the Colorado potato beetle and changes in the midgut and peritrophic membrane"],"prefix":"10.1002","volume":"78","author":[{"given":"Dongdong","family":"Liu","sequence":"first","affiliation":[{"name":"Laboratory of Agrozoology, Department Plants and Crops, Faculty of Bioscience Engineering Ghent University  Ghent 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