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Insight into the dynamics of Ixodes ricinus nymphal midgut proteome

ABSTRACT: The midgut of the tick is the most important tissue for the storage and digestion of host blood, which serves as the sole source of energy and nutrients for all tick development and reproduction. During feeding at each developmental stage, dynamic changes in the tick midgut epithelium reflect changes in physiological processes in this tissue. In addition, the midgut serves as the primary interface between the tick and tick-borne pathogens, which determines the vector competence of the tick. Several transcriptome data from Ixodes ricinus have been published, but few studies have examined tick proteomes. In contrast to transcriptome data, proteomics provides a deeper understanding of key cellular processes occurring in the investigated tissues. In this work, we present for the first time insight into proteomics of the midgut of I. ricinus nymph. Label-free quantitative proteomics was used to elucidate changes during blood meal and development in I. ricinus. A total of 1 534 I. ricinus-specific proteins were identified, with only a few host proteins detected. In addition, the proteins involved in the specific physiological processes of the tick were studied in detail to gain better insight into the ontogeny of the midgut of the nymph.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Ixodes Ricinus

TISSUE(S): Midgut

SUBMITTER: Filip Dyčka

LAB HEAD: Petr Kopáček

PROVIDER: PXD042405 | Pride | 2023-11-10

REPOSITORIES: Pride

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Project description:Ticks are obligate hematophagous arthropods that transmit a wide range of pathogens to humans as well as wild and domestic animals. They also harbor a non-pathogenic microbiota, although our previous study has shown that the diverse bacterial microbiome in the midgut of Ixodes ricinus is quantitatively poor and lacks a core microbe. In artificial infections by capillary feeding of ticks with two model bacteria (Gram-positive Micrococcus luteus and Gram-negative Pantoea sp.), rapid clearance of these microbes from the midgut was observed, indicating the presence of active immune mechanisms in this organ. In the current study, RNA-seq analysis was performed on the midgut of I. ricinus females inoculated with either M. luteus or Pantoea sp. or with sterile water as a control. While no immune-related transcripts were upregulated by microbial inoculation compared to the sterile control, capillary feeding itself triggered dramatic transcriptional changes in the tick midgut. Manual curation of the transcriptome from the midgut of unfed I. ricinus females, complemented by proteomic analysis, revealed the presence of several constitutively expressed putative antimicrobial peptides (AMPs) that are independent of microbial stimulation and are referred to here as ‘guard’ AMPs. These included two types of midgut-specific defensins, two different domesticated amidase effector 2 (Dae2), microplusin/ricinusin-related molecules, two lysozymes and two gamma interferon-inducible lysosomal thiol reductases (GILTs). The in vitro antimicrobial activity assays of two synthetic mature defensins, defensin 1 and defensin 8, confirmed their specificity against Gram-positive bacteria showing exceptional potency to inhibit the growth of M. luteus at nanomolar concentrations. The antimicrobial activity of midgut defensins is likely part of a multicomponent system responsible for the rapid clearance of bacteria in the tick midgut. Further studies are needed to evaluate the role of other identified ‘guard‘ AMPs in controlling microorganisms entering the tick midgut.

			Action	DRS
	210607_14AD.tar.gz	Other
	210607_2D.tar.gz	Other
	210607_4wAD.tar.gz	Other
	210607_FF.tar.gz	Other
	210607_UF.tar.gz	Other

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Insight into the dynamics of Ixodes ricinus nymphal midgut proteome

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