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      Embryonic hematopoiesis modulates the inflammatory response and larval hematopoiesis in Drosophila

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          Abstract

          Recent lineage tracing analyses have significantly improved our understanding of immune system development and highlighted the importance of the different hematopoietic waves. The current challenge is to understand whether these waves interact and whether this affects the function of the immune system. Here we report a molecular pathway regulating the immune response and involving the communication between embryonic and larval hematopoietic waves in Drosophila. Down-regulating the transcription factor Gcm specific to embryonic hematopoiesis enhances the larval phenotypes induced by over-expressing the pro-inflammatory Jak/Stat pathway or by wasp infestation. Gcm works by modulating the transduction of the Upd cytokines to the site of larval hematopoiesis and hence the response to chronic (Jak/Stat over-expression) and acute (wasp infestation) immune challenges. Thus, homeostatic interactions control the function of the immune system in physiology and pathology. Our data also indicate that a transiently expressed developmental pathway has a long-lasting effect on the immune response.

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          Most cited references75

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          Cytokine/Jak/Stat signaling mediates regeneration and homeostasis in the Drosophila midgut.

          Cells in intestinal epithelia turn over rapidly due to damage from digestion and toxins produced by the enteric microbiota. Gut homeostasis is maintained by intestinal stem cells (ISCs) that divide to replenish the intestinal epithelium, but little is known about how ISC division and differentiation are coordinated with epithelial cell loss. We show here that when enterocytes (ECs) in the Drosophila midgut are subjected to apoptosis, enteric infection, or JNK-mediated stress signaling, they produce cytokines (Upd, Upd2, and Upd3) that activate Jak/Stat signaling in ISCs, promoting their rapid division. Upd/Jak/Stat activity also promotes progenitor cell differentiation, in part by stimulating Delta/Notch signaling, and is required for differentiation in both normal and regenerating midguts. Hence, cytokine-mediated feedback enables stem cells to replace spent progeny as they are lost, thereby establishing gut homeostasis.
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            Identification of in vivo DNA targets of chromatin proteins using tethered dam methyltransferase.

            We have developed a novel technique, named DamID, for the identification of DNA loci that interact in vivo with specific nuclear proteins in eukaryotes. By tethering Escherichia coli DNA adenine methyltransferase (Dam) to a chromatin protein, Dam can be targeted in vivo to native binding sites of this protein, resulting in local DNA methylation. Sites of methylation can subsequently be mapped using methylation-specific restriction enzymes or antibodies. We demonstrate the successful application of DamID both in Drosophila cell cultures and in whole flies. When Dam is tethered to the DNA-binding domain of GAL4, targeted methylation is limited to a region of a few kilobases surrounding a GAL4 binding sequence. Using DamID, we identified a number of expected and unexpected target loci for Drosophila heterochromatin protein 1. DamID has potential for genome-wide mapping of in vivo targets of chromatin proteins in various eukaryotes.
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              G-TRACE: rapid Gal4-based cell lineage analysis in Drosophila

              We combine Gal4/UAS, FLP/FRT and fluorescent reporters to generate cell clones that provide spatial, temporal, and genetic information about the origins of individual cells in Drosophila. We name this combination the Gal4 Technique for Real-time and Clonal Expression (G-TRACE). The approach should allow for screening and the identification of real-time and lineage-traced expression patterns on a genomic scale.
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                Author and article information

                Contributors
                Role: Reviewing Editor
                Journal
                eLife
                Elife
                eLife
                eLife
                eLife Sciences Publications, Ltd
                2050-084X
                11 July 2018
                2018
                : 7
                : e34890
                Affiliations
                [1 ]Institut de Génétique et de Biologie Moléculaire et Cellulaire IllkirchFrance
                [2 ]deptUMR7104 Centre National de la Recherche Scientifique IllkirchFrance
                [3 ]deptU1258 Institut National de la Santé et de la Recherche Médicale IllkirchFrance
                [4 ]Université de Strasbourg IllkirchFrance
                [5]University of California, Los Angeles United States
                [6]University of California, Los Angeles United States
                Author notes

                These authors contributed equally to this work.

                [†]

                These authors contributed equally to this work.

                Author information
                https://orcid.org/0000-0001-5301-1975
                http://orcid.org/0000-0001-6278-5120
                Article
                34890
                10.7554/eLife.34890
                6040882
                29992900
                e71557d4-f11f-4d2f-9c63-4450ba6825e4
                © 2018, Bazzi et al

                This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

                History
                : 08 January 2018
                : 18 June 2018
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001677, Institut National de la Santé et de la Recherche Médicale;
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100004794, Centre National de la Recherche Scientifique;
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100004099, Ligue Contre le Cancer;
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100004097, Fondation ARC pour la Recherche sur le Cancer;
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100007379, Fondation pour l'Aide à la Recherche sur la Sclérose en Plaques;
                Award ID: Research grant and postdoc fellowship
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100003768, Université de Strasbourg;
                Award ID: Graduate student fellowship
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100001665, Agence Nationale de la Recherche;
                Award ID: ANR-10-LABX-0030-INRT
                Award Recipient :
                The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
                Categories
                Research Article
                Developmental Biology
                Stem Cells and Regenerative Medicine
                Custom metadata
                Distinct hematopoietic waves interact via transcriptional cascades specific to embryonic hemocytes to ensure the proper immune response in Drosophila.​.

                Life sciences
                drosophila,hematopoiesis,jak/stat,inflammation,transcription factors,gcm,d. melanogaster
                Life sciences
                drosophila, hematopoiesis, jak/stat, inflammation, transcription factors, gcm, d. melanogaster

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