Steinernema carpocapsae Breton small RNA sequencing insect in vitro simulation system, from "The genome, transcriptome, and proteome of the nematode Steinernema carpocapsae: evolutionary signatures of a pathogenic lifestyle"
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ABSTRACT:
ORGANISM(S): Steinernema carpocapsae
SUBMITTER: José Bermúdez-Barrientos
PROVIDER: E-GEOD-85256 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:We report small RNA sequencing of the entomopathogenic nematode Steinernema carpocapsae. The nematodes were grown in liquid culture in homogenates of pig kidney/fat and infective juveniles were gathered. Then Galleria mellonella insect haemolymph was added to simulate insect infection, control nematodes weren't added haemolymph. Nematodes were collected after two hours after haemolymph addition.
2016-09-05 | GSE85256 | GEO
Project description:Entomopathogenic nematodes (Rhabditida; Steinernematidae and Heterorhabditidae) are insect parasites which are of economic importance due to their use as biocontrol agents. The third larval stage, infective juveniles (IJs) leave the natal host and move in the soil to locate a new insect host. Both families are associated with mutualistic bacteria, which are released into the haemolymph of the host insect, killing the insect and providing nutrition for nematode development and reproduction, and are thus categorised as entomopathogenic nematodes. Similarities between these two families are due to convergent evolution associated with this lifestyle, rather than common ancestry. Heterorhabditids are closely related to the vertebrate parasites Strongylida (Adams and Nguyen, 2002) and to Caenorhabditis elegans, whereas steinernematids are more closely related to Strongyloididae. The IJ stage of parasitic nematodes is analogous to the dauer stage of C. elegans; both are developmentally arrested, stress resistant stages that disperse to colonise new hosts or food resources, respectively. By investigating the molecular mechanisms and consequences of temperature acclimation and aging in EPN IJs, insights into survival and the changes induced by low temperature exposure may be gained. Label free quantitative (LFQ) massspectrometery-based proteomics facilitates the identification and quantification of thousands of proteins in a single run. Such data allows for the comparison of the proteomes of EPN IJs after conditioning via gene ontology mapping and functional enrichment analysis. Understanding how the IJ proteome is affected by temperature and aging can provide a molecular basis for the wide array of phenotypes and behaviours these organisms may adopt. This study aims to provide proteomic data which may elucidate the molecular mechanisms underlying the phenotypic plasticity which EPN IJs of two distantly related species exhibit.
2022-03-27 | PXD027608 | Pride
Project description:Entomopathogenic nematode infective juveniles activate when they infect a host and begin releasing excreted-secreted products. These are the excreted-secreted proteins detected by mass spec from activated Steinernema feltiae infective juveniles. The infective juveniles were activated in vitro by exposure to insect-host tissue for 6 hours.
2018-10-02 | MSV000082993 | MassIVE
Project description:The spore-forming bacterium Bacillus thuringiensis (Bt) of B. cereus group uses toxin-opened breaches at the insect midgut epithelium to infest the haemolymph where it rapidly propagates despite antimicrobial host defences to induce eventually host death by acute septicaemia. The response of Bt to haemolymph and the latter’s role in bacterial pathogenesis is an area that needs clarification. We report the proteome analysis of Bt subsp. kurstaki (Btk) haemolymph stimulon consisted of significant changes in 60 (34 up- and 26 down-regulated) differentially accumulated proteins (DAPs). Gene Ontology (GO) enrichment analysis revealed Btk haemolymph-responsive DAPs were mainly involved in glutamate metabolism, transketolase activity and ATP-dependent transmembrane transporters. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis disclosed that DAPs were highly enriched in enzymes related to the biosynthesis of ansamycins, a family of bacterial polyketides with antimicrobial activity. Interestingly, around 30% of DAPs were putative virulence factors, including polyketide synthase, ABC transporter, and cytotoxin. Collectively, our findings suggest Btk haemolymph stimulon could be partially driven bacterial survival and propagation within the haemolymph of infected insects, contributing to its remarkable success as entomopathogen.
2021-11-02 | PXD021830 | Pride
Project description:Entomopathogenic nematodes (EPNs) of the genera Heterorhabditis are obligate and lethal insect parasites. In recent years they have been used increasingly as biological control agents. These EPNs are symbiotically associated with bacteria of the genera Photorhabdus. The bacterial symbionts are essential to kill the host (within 24-48 hours) and digest its tissues to provide nutrients for themselves as well for expanding nematodes. Drosophila larvae are suitable insect hosts and part of the tripartite model system we used before to show the importance of haemolymph clotting and eicosanoids during the infection. We used the well-established tripartite model (Drosophila, nematodes, bacteria), DNA chips and bioinformatic tools to compare gene expression in non-infected and infected fly larvae. We focused on the early time point of nematode infection and therefore infected Drosophila larvae using H. bacteriophora harbouring GFP-labelled P. luminescens bacteria. Infected (GFP positive) larvae were collected 6 hours after infection.
2014-03-07 | E-MTAB-1542 | biostudies-arrayexpress
Project description:Entomopathogenic nematodes (EPNs) are unique parasitic nematodes due to their symbiosis with entomopathogenic bacteria and their ability to kill insect hosts quickly after infection. Although it has been widely believed that EPNs rely on their bacterial partners for killing insect hosts, compelling evidence from previous studies challenges this model. We developed an improved method of activating millions of Steinernema carpocapsae infective juveniles (IJs) in vitro to harvest excreted/secreted (ES) proteins for bioactivity tests and proteomics analysis. We found that a low dose of the ES proteins from early activated nematodes is lethal to Drosophila melanogaster adults within 2-6 hours. We analyzed the protein composition of this venom using mass spectrometry and identified 472 proteins. Many of these venom proteins share high homology with those of vertebrate-parasitic nematodes. Among many different families of proteins found in the venom, proteases and protease inhibitors are especially abundant. Some toxin-related proteins such as Shk domain-containing proteins were also detected. We further analyzed the transcriptomes of individual non-activated IJs and nematodes that were activated in vitro and in vivo, which revealed a dramatic shift in gene expression during IJ activation. By comparing the whole transcriptomes and the genes encoding venom proteins between the in vitro and in vivo activated nematodes, we confirmed that the in vitro activation is a good approximation of the in vivo process. In summary, our findings strongly support a new model that S. carpocapsae and likely other Steinernema EPNs have a more active role in contributing to the pathogenicity of the nematode-bacterium complex than simply relying on their symbiotic bacteria. Furthermore, we propose that EPNs are a good model system for investigating vertebrate- and human-parasitic nematodes, especially regarding the function of ES products.
2017-03-27 | GSE89961 | GEO
Project description:Diapause is a key life-history event characterised by arrested development, suppressed metabolism and increased stress tolerance. This state allows an organism to avoid prolonged periods of harsh and inhospitable environmental conditions. For species where only mated female adults undergo diapause, mating is performed prior to diapause. Mating can have a profound effect on the behaviour and physiology of females resulting in changes to the expression of key biological processes, including immunity. However, our understanding of how mating impacts long-term immunity and whether these effects persist throughout diapause is currently limited. Here we explored proteomic changes in the haemolymph of the ecologically important pollinator, Bombus terrestris. B. terrestris queens mate prior to diapause (a non-feeding arrest of development that can last 6-9 months). Using mass-spectrometry-based proteomics, we quantified changes in the pre-diapause queen haemolymph after mating, as well as the subsequent protein expression of mated queens during and post-diapause. Our results provide clear molecular evidence for the consequences and benefits of mating at the immune level through the selective increased abundance of antimicrobial peptides that are sustained throughout diapause. In addition our results provide novel insights into the molecular mechanisms by which bumblebees prepare for, survive, and recover from diapause, insights that may have implications for our general understanding of these processes in other insect groups.
2019-11-18 | PXD014384 | Pride
Project description:Enabled by recently available genomic resources for bumblebees, nano LC-MS/MS based proteomics was conducted to characterise the queen haemolymph proteome. The resulting mass spectrometry data were also utilised in a proteogenomic capacity to identify proteins missing from official gene sets and to improve gene models and annotation. 97 multiple-peptide supported proteins within queen bumblebee haemolymph were identified including 28 immunity-related proteins with roles in immune recognition, signalling and regulation. In addition 19 proteins with potential immune roles were identified. The haemolymph proteome is particularly rich in metabolism and transport proteins and comprised 18 proteins with unknown function.
2015-02-24 | PXD001644 | Pride
Project description:Very little is known about the genetic modulation of nictation, a host-finding strategy used by infective juveniles of many pathogenic nematodes, as well as a phoretic strategy of Caenorhabditis elegans dauers. We here use a mass spectrometry-driven approach to find neuropeptides involved in the modulation of this behavior. We identified 126 neuropeptides in infective juveniles of the commercially relevant entomopathogenic nematode Steinernema carpocapsae, 75 of which have orthologs in the model organism C. elegans. To prioritize candidates for causality testing in light of the stage-restricted nictation behavior, we developed quantitative peptidomic assays and carried out targeted proteomic measurements that allows comparing neuropeptide levels at different C. elegans life stages. Our results show that virtually all 164 quantified neuropeptides are more abundant in dauers compared to L3 juveniles. Even so, not all are relevant to nictation, and we identify the neuropeptide genes flp-7 and flp-11 as novel regulators of nictation.
2022-12-08 | PXD034629 | Pride
Project description:We analyzed the consequences of recombination in gene expression. For this, we measured expression patterns in two C. elegans wild types, N2 and CB4856 (CB). We compare these profiles with gene expression values from a Recombinant Inbred Population (RIL) derived from the same wild types. Expression values of the RILs (GSE17071) were obtained from Viñuela et al. (Genome Research, 2010). N2 and CB strain nematodes were cultured in identical conditions. Likewise, mRNA was extracted at three different ages (juveniles, reproductive and old worms) to investigate the transcriptional consequences of recombination in aging worms. Then, we explored gene expression heritability and transgression as genetic parameters for the analysis of gene expression divergence in natural isolates. Moreover, we investigated the progression of those parameters with age. In total, 14 dual-color microarrays were done on two wild type strains (N2 and CB4856) and three age groups (40, 96, and 214 hours). 5 replicates of t1 (juvenile) and t3 (senescent) samples, and 4 replicates of t2 (reproductive) samples, in a dye-swap design.
2012-03-31 | E-GEOD-22887 | biostudies-arrayexpress