Analysis of the humoral immune response against aberrant self by RNA sequencing of the fat body in Drosophila melanogaster
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ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE41487 | GEO | 2014/03/07
SECONDARY ACCESSION(S): PRJNA177218
REPOSITORIES: GEO
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Project description:The early stages of the immune response against aberrant tissues such a tumors are technically difficult to study and therefore little understood. To analyze the innate immune response towards modified self-tissue we expressed a dominant-active version of the Ras oncogene in Drosophila imaginal discs and salivary glands. Signs of an immune response are observed on wings of adult flies, which show spots of melanization and in larvae, where the salivary glands attract blood cells. In line with the strong response mounted against salivary glands, they are found to express metalloproteases and apoptotic markers making them both accessible and recognizable for the immune system. The response against the glands bears several hallmarks of an encapsulation response including flattening of plasmatocytes, differentiation of lamellocytes, attachment of crystal cells and melanization at the final stage. RNA sequencing of the fat body, the primary producer of immune effectors, reveals a pattern of induced genes, which overlaps the one previously found upon wasp infestation but shows also several specific features. Evidence for the functional importance of one of the induced genes is presented proving the power of our approach to identify regulators of the response against modified self. Transcriptome analysis of the immune response in the fat body after expression of dominant-active RAS in the wing imaginal discs and salivary glands. Comparison of fat bodies from 3 control and 3 RAS-expressing larvae. For each sample fat bodies from 40 individuals were dissected. Differential gene expression files [output of Cuffdiff] are linked below: ResultsContr123vsRas123Individual.xls: Output file of Cuffdiff. Differential gene expression analysis, three control compared to individual Ras induced; replicate analysis using Cuffdiff. ResultsContr123vsRas123Multiple.xls: Output file of Cuffdiff. Differential gene expression analysis, three control compared to three Ras induced; replicate analysis using Cuffdiff. ResultsContr123vsRas12Multiple.xls: Output file of Cuffdiff. Differential gene expression analysis, three control compared to two Ras induced; replicate analysis using Cuffdiff.
2014-03-07 | E-GEOD-41487 | biostudies-arrayexpress
Project description:In Drosophila melanogaster larval hemolymph, under normal conditions, plasmatocytes and crystal cells represent respectively ~95% and ~5% of hemocytes, while lamellocytes, the third larval cell type, are absent since they are only induced after parasitoid wasp oviposition, their role being the encapsulation-melanization response to eliminate the wasp egg. However, even after induction lamellocytes number remains low, making difficult biochemical studies. Here using the D. melanogaster hopTum-l mutant that constitutively produces a high number of hemocytes, we set up a method to purify lamellocytes and analyzed their major proteins by 2D gel electrophoresis and their biotinylated plasma membrane surface proteins by 1D SDS-PAGE after affinity purification. Mass spectrometry allowed to identify 430 proteins from the 2D spots and 344 from affinity purified proteins, totalizing 639 unique proteins. Known lamellocyte markers such as PPO3 and the integrin myospheroid are among the major proteins and affinity purification led to the detection of other integrins and a large array of integrins associated proteins involved in cell-cell junction formation and function. Overall newly identified proteins indicated that these cells are highly adapted to the encapsulation process but may have also several different physiological functions. This study provides the basis for new lamellocyte studies in vivo and in vitro, and develop markers to search whether different populations coexist, establish their origins and decipher their respective roles in drosophila physiology and immunity.
2024-04-29 | PXD016876 | Pride
Project description:Epithelial and especially mucosal immunity represents the first line of defence against the plethora of potential pathogens trying to invade via the gastrointestinal tract. The salivary glands of the fruit fly are an indispensable part of the gastrointestinal tract, but their contribution to the mucosal immunity has almost completely been neglected. Our major goal was to elucidate if the fly's salivary glands are able to mount an immune response and what the major characteristics of this immune response are.
2010-03-18 | GSE20938 | GEO
Project description:Postmitotic tissues are incapable of replacing damaged cells through proliferation, but need to rely on buffering mechanisms to prevent tissue disintegration. By constitutively activating the Ras/MAPK-pathway via RasV12-overexpression in the postmitotic salivary glands of Drosophila larvae, we overrode the glands adaptability to growth signals, induced hypertrophy and stress accumulation. This allowed us to decipher a novel, spatio-temporally regulated interaction between the JNK-stress response and a genuine tissue-autonomous immune response. Central to this interaction is the direct inhibition of JNK-signalling by the antimicrobial peptide Drosomycin, which blocks programmed cell death and prevents recognition of the stressed tissue by the systemic immune response. While this mechanism might allow growing salivary glands to cope with temporary stress, continuous expression of Drosomycin favors survival of unrestricted, hypertrophic RasV12-glands. Our findings indicate the necessity for refined therapeutic approaches that fundamentally acknowledge detrimental effects that stimulated immune responses have on tissues coping with damage and stress.
2020-12-30 | GSE138936 | GEO
Project description:Studying the salivary glands potential role during intestinal infections,using advantage from the ingenious GAL4/UAS-system available in the fly Epithelial immunity is a very simple but nevertheless indispensable type of immune response. Amongst all epithelial tissues, the intestine and the airways are outstanding, because they provide huge surface areas, where colonization or invasion of potential pathogens has to be obviated. The intestine is of special interest, because it has to hold the balance between tolerance and immunity, to protect the own microbial flora. An essential part of the intestinal tract has been completely overlooked, the salivary glands. They are the gatekeepers of the intestinal system, being essential for various aspects of the intestineâs immunity. Using the bipartite Gal4/UAS expression system, it is possible to activate the immune system ectopically in different organs of the fly. Activation of the IMD-pathway in the salivary glands is possible; because a very specific driver line is available that directs expression of any gene of interest into the salivary glands only. Flies, where the IMD-pathway in the salivary glands has been activated have a very distinct phenotype. They show a smaller body length as well as a smaller salivary gland length than the parental animals Microarray data analysis showed that 457 genes were upregulated and 578 genes downregulated. Interestingly, the sets of regulated genes show only a very small overlap with the canonical set of Drosophila immune genes. Other physiological scenarios such as autophagic cell death are apparently also not activated upon IMD-pathway activation. Among the regulated genes, those that code for signaling associated protease activity are significantly modulated. This holds especially true for presenilin and the signal peptide peptidase. The comparison of the transcriptional events induced following IMD-activation in the trachea and the salivary glands shows also only a small overlap, indicating that the general IMD-activated core transcriptome is rather small. In conclusion, the salivary glands may a very good tool to study the physiological role of selected genes that are of importance for human health such as the Alzheimerâs disease related presenilin gene in a functional environment. Ectopic activiation of the Immune deficiency pathway in the larval salivary glands using the Gal4/UAS-system of Brand and Perrimon. The activation was induced by the overexpression of the pattern recognition receptor PGRP-LCx using a driver line which was specific for the salivary glands.In general four replicates were performed including dye-swaps in two-colour arrays.
2010-05-05 | E-GEOD-20938 | biostudies-arrayexpress
Project description:In spite of the many recent developments in the field of vector sialomics, the salivary glands of larvalmosquitoes have been largely unexplored. We used whole-transcriptome microarray analysis to create a gene-expression profile of the salivary gland tissue of fourth-instar Anopheles gambiae larvae, and compare it to the gene-expression profile of a matching group of whole larvae. We identified a total of 221 probes with expression values that were (a) significantly enriched in the salivary glands, and (b)sufficiently annotated as to allow the prediction of the presence/absence of signal peptides in their corresponding gene products. Based on available annotation of the protein sequences associated with these probes, we propose that the main roles of larval salivary secretions include: (a) immune response, (b) mouthpart lubrication, (c) nutrient metabolism, and (d) xenobiotic detoxification. Other highlights of the study include the cloning of a transcript encoding a previously unknown salivary defensin (AgDef5), the confirmation of mucus secretion by the larval salivary glands, and the first report of salivary lipocalins in the Culicidae. Keywords: Anopheles gambiae, salivary gland, Diptera, gene expression, salivary defensin, transcriptome, salivary lipocalin
2008-05-01 | GSE9642 | GEO
Project description:Although the salivary transcriptome of adult mosquitoes has been thoroughly described in several recent papers, very little information exists regarding the biological role of larval salivary glands in the Culicidae. We used whole-transcriptome Affymetrix® chips to compare the transcriptional profiles of Anopheles gambiae larval (L4) salivary glands and whole larvae. We identified a total of 277 transcripts as being significantly enriched in the salivary glands. Based on available annotation for the known or predicted protein sequences encoded by these transcripts, 41 were identified as corresponding to secreted proteins, 233 as corresponding to non-secreted (housekeeping) proteins, and 3 as encoding proteins of unknown function. Based on functional annotation of the putatively secreted gene products, we propose that larval salivary secretions have roles in nutrient digestion, detoxification, immunity, and mouthpart lubrication. Interestingly, several components of the larval saliva (e.g. apyrase and serine proteases) have also been reported to exist in adult female saliva, where they are though to help regulate a vertebrate hostâs immune response to bloodfeeding. In conclusion, our results suggest that the salivary glands are important components of both the digestive and immune systems of larval mosquitoes, and that their study might provide clues about the evolution of adaptations to bloodfeeding observed in adults. Experiment Overall Design: We attempted to identify transcripts that are both present and significantly enriched in the salivary glands (SG) of the 4th instar Anopheles gambiae larva. Transcripts were considered as present in the SG if the built-in Affymetrix detection call scored them as such in all three biological replicates of the experiment. Expression values (i.e. signal intensities) of transcripts from the salivary glands were compared to those of a matching group of whole 4th instar larvae using a t-test. Transcripts showing an expression value two-fold or higher (p < 0.01; false discovery rate=1%) in the SG as compared to whole larvae were considered to be significantly enriched.
2008-11-06 | E-GEOD-9642 | biostudies-arrayexpress
Project description:Simultaneous analysis of Plasmodium falciparum and Anopheles gambiae gene expression: an assessment of parasite maturation and of the mosquito response to infection. Malaria is probably the most lethal and most prevalent parasitic disease in the world, causing more than one million deaths per year. In spite of intensive research, no vaccine exists and good vaccine candidates are still a necessity. Plasmodium sporozoites from mosquito midguts are not as infective as those from salivary glands, presenting a different type of motility which is presumably associated with their invasive capacity. It is assumed that these differences are due to maturation of sporozoites, which happens either in the salivary glands or somewhere along the way from the midgut to the glands. Indeed, the invasion of the salivary gland is considered to be an essential step for malaria transmission to the host and it is known to causes expression of multiple genes in the parasite and activation of some immune-response genes in the insect. Here we compared the gene expression profile of P. falciparum parasites from midgut and salivary glands in order to search for genes involved in parasite maturation; on the assumption that any differentially expressed genes found under this condition could be potentially related to the higher infectivity of the salivary gland derived sporozoites and therefore would be potentially good vaccine candidates. Also, because very little is known about mosquito genes involved in the response to Plasmodium, we compared the gene expression profile of infected and uninfected midguts and salivary glands. Our results demonstrate that, upon infection, at least 13 genes are differentially expressed in the mosquito midgut and 43 in the salivary gland. Gene ontology enrichment analysis showed that among other things, in mosquito salivary glands, infection causes multiple genes related to chitin metabolism to be differentially expressed. Also and more importantly, at least 54 genes were differentially expressed in P. falciparum from salivary glands in comparison to midgut (19 down-regulated and 35 up-regulated). Last, we show for the first time that P. falciparum causes multiple genes in the salivary gland to be regulated and that some of those genes could be potentially beneficial to the parasite.
2014-10-01 | GSE18780 | GEO
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular functions. The goal of this study is to compare NGS-derived salivary gland transcriptome profilings (RNA-seq) to better understand the molecular nature of the physiological differences in adult murine salivary glands. Methods: Major murine salivary gland mRNA profiles were generated by deep sequencing, in triplicate, using Illumina HiSeq 2000. The sequence reads that passed quality filters were analyzed at the gene level with STAR followed by Cufflinks. In vivo NaCl reabsorption measurements were performed for validation. Results: Using an optimized data analysis workflow, we mapped about 15 million sequence reads per sample to the mouse genome (build mm10) and identified 1991 genes that were differentially expressed across three major salivary glands. RNA-seq data provided valuable insights into the nature of the functional differences among the major salivary glands Conclusions: Our study represents the first detailed analysis of murine salivary gland transcriptomes, with biologic replicates, generated by RNA-seq technology. Our results confirm functions of many genes, identified using genetically modified mice. We conclude that RNA-seq-based transcriptome characterization would offer a comprehensive and sensitive evaluation of the gene expression.
2018-02-28 | GSE96747 | GEO
Project description:Over the last 20 years, the advances in sequencing technologies highlighted the unique composition of the salivary glands of blood-feeding arthropods. Further biochemical and structural data demonstrated that salivary proteins can disrupt host hemostasis, inflammation and immune response in addition to favor pathogen transmission. Previously, a Sanger-based sialome of the adult Ochlerotatus. triseriatus female salivary glands was published based on 731 ESTs. Here we revisited O. triseriatus salivary glands contents using an Illumina-sequencing approach of both male and female tissues. In the current data set we report 10,317 coding DNA sequences classified into several functional classes. The translated transcripts also served as reference database for the proteomic analysis of O. triseriatus female saliva, in which unique peptides of 101 proteins were found. Finally, comparison of male and female libraries allowed the identification of female-enriched transcripts that are potentially related to blood acquisition and virus transmission.
2022-10-15 | PXD033588 | Pride