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3. Author response: Dynamics of macrophage polarization support Salmonella persistence in a whole living organism

Author

Leiba, Jade, primary, Sipka, Tamara, additional, Begon-Pescia, Christina, additional, Bernardello, Matteo, additional, Tairi, Sofiane, additional, Bossi, Lionello, additional, Gonzalez, Anne-Alicia, additional, Mialhe, Xavier, additional, Gualda, Emilio J, additional, Loza-Alvarez, Pablo, additional, Blanc-Potard, Anne, additional, Lutfalla, Georges, additional, and Nguyen-Chi, Mai E, additional

Published
2023
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6. The IbeA protein from adherent invasive Escherichia coli is a flavoprotein sharing structural homology with FAD‐dependent oxidoreductases.

Author

Paris, Théo, Kiss, Agneta, Signor, Luca, Lutfalla, Georges, Blaise, Mickaël, Boeri Erba, Elisabetta, Chaloin, Laurent, and Yatime, Laure

Subjects
LIGAND binding (Biochemistry), ESCHERICHIA coli, FLAVOPROTEINS, FLAVIN adenine dinucleotide, OXIDOREDUCTASES, CROHN'S disease, FLUORESCENCE spectroscopy
Abstract

Invasion of brain endothelium protein A (IbeA) is a virulence factor specific to pathogenic Escherichia coli. Originally identified in the K1 strain causing neonatal meningitis, it was more recently found in avian pathogenic Escherichia coli (APEC) and adherent invasive Escherichia coli (AIEC). In these bacteria, IbeA facilitates host cell invasion and intracellular survival, in particular, under harsh conditions like oxidative stress. Furthermore, IbeA from AIEC contributes to intramacrophage survival and replication, thus enhancing the inflammatory response within the intestine. Therefore, this factor is a promising drug target for anti‐AIEC strategies in the context of Crohn's disease. Despite such an important role, the biological function of IbeA remains largely unknown. In particular, its exact nature and cellular localization, i.e., membrane‐bound invasin versus cytosolic factor, are still of debate. Here, we developed an efficient protocol for recombinant expression of IbeA under native conditions and demonstrated that IbeA from AIEC is a soluble, homodimeric flavoprotein. Using mass spectrometry and tryptophan fluorescence measurements, we further showed that IbeA preferentially binds flavin adenine dinucleotide (FAD), with an affinity in the one‐hundred nanomolar range and optimal binding under reducing conditions. 3D‐modeling with AlphaFold revealed that IbeA shares strong structural homology with FAD‐dependent oxidoreductases. Finally, we used ligand docking, mutational analyses, and molecular dynamics simulations to identify the FAD binding pocket within IbeA and characterize possible conformational changes occurring upon ligand binding. Overall, we suggest that the role of IbeA in the survival of AIEC within host cells, notably macrophages, is linked to modulation of redox processes. [ABSTRACT FROM AUTHOR]

Published
2024
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7. Dynamics of macrophage polarization support Salmonella persistence in a whole living organism

Author

Leiba, Jade, primary, Sipka, Tamara, additional, Begon-Pescia, Christina, additional, Bernardello, Matteo, additional, Tairi, Sofiane, additional, Bossi, Lionello, additional, Gonzalez, Anne-Alicia, additional, Mialhe, Xavier, additional, Gualda, Emilio, additional, Loza-Alvarez, Pablo, additional, Blanc-Potard, Anne, additional, Lutfalla, Georges, additional, and Nguyen-Chi, Mai, additional

Published
2023
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12. Ontogenetic Changes in Blood Osmolality During the Postembryonic Development of Zebrafish ( Danio rerio )

Author

Charmantier, Guy, Nguyen-chi, Mai, Lutfalla, Georges, Charmantier, Guy, Nguyen-chi, Mai, and Lutfalla, Georges

Abstract

The zebrafish Danio rerio is a teleost model species widely used in developmental genetics, biomedical studies, toxicology, and drug screening. Despite the interest of this species in research, little is known through indirect observations about its blood osmolality, which is a key parameter for diverse experiments. In this study, we directly measured blood osmolality using nano-osmometry at different stages of zebrafish postembryonic development. We found that blood osmolality is close to 240 mOsm·kg−1 in early larvae. It progressively increased to ∼270 mOsm·kg−1 during the larval development before reaching ∼300 mOsm·kg−1 after metamorphosis in juveniles and later in adults. These ontogenetic changes in blood osmolality illustrate the physiological changes in osmoregulation associated with postembryonic development, including metamorphosis. These values are of practical interest for adjusting the osmolality of fixatives and cell and tissue culture media for research using zebrafish as a model.

Published
2022
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18. ProtectinD1_controls_immune_cells_during_regeneration_NguyenChi_et_al_2020_AuthorsVersion

Author

Nguyen-Chi, Mai, Luz-Crawford, Patricia, Balas, Laurence, Sipka, Tamara, Contreras-López, Rafael, Barthelaix, Audrey, Lutfalla, Georges, Durand, Thierry, Jorgensen, Christian, and Djouad, Farida

Subjects
fungi, Regeneration – macrophage – neutrophil – inflammation – specialized pro-resolving lipid mediators - zebrafish
Abstract

Specialized pro-resolving mediators (SPM) are a family of lipids controlling the resolution of inflammation and playing a role in many processes including organ protection and tissue repair. While synthetic SPM are potent bioactive molecules in vivo, their role in epimorphic regeneration of organs in vertebrates has never been proved. Using the zebrafish larvae as a robust regenerative vertebrate system, we show that the SPM neuroprotectin/protectin D1 (PD1) improves the regeneration of the zebrafish caudal fin. Acting in a narrow time window during regeneration, PD1 accelerates the resolution of inflammation without affecting the initial kinetic of neutrophil infiltration but instead, promoting their reverse migration potential. In addition, PD1 induces macrophage polarization switch toward non-inflammatory states in both zebrafish and mammalian system. Finally, macrophages but not neutrophils are essential for PD1-mediated regeneration. Thus, our findings strongly support the development of pro-resolving mediators as promising natural therapeutic candidates for inflammatory and degenerative disorders.

Published
2021
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20. Occurrence of foamy macrophages during the innate response of zebrafish to trypanosome infections

Author

Jacobs, Sem H., Dóró, Eva, Hammond, Ffion R., Nguyen-Chi, Mai E., Lutfalla, Georges, Wiegertjes, Geert F., Forlenza, Maria, Jacobs, Sem H., Dóró, Eva, Hammond, Ffion R., Nguyen-Chi, Mai E., Lutfalla, Georges, Wiegertjes, Geert F., and Forlenza, Maria

Abstract

A tightly regulated innate immune response to trypanosome infections is critical to strike a balance between parasite control and inflammation-associated pathology. In this study, we make use of the recently established Trypanosoma carassii infection model in larval zebrafish to study the early response of macrophages and neutrophils to trypanosome infections in vivo. We consistently identified high-and low-infected individuals and were able to simultaneously characterise their differential innate response. Not only did macrophage and neutrophil number and distribution differ between the two groups, but also macrophage morphology and activation state. Exclusive to high-infected zebrafish, was the occurrence of foamy macrophages characterised by a strong pro-inflammatory profile and potentially associated with an exacerbated immune response as well as susceptibility to the infection. To our knowledge, this is the first report of the occurrence of foamy macrophages during an extracellular trypanosome infection.

Published
2021

30. Genome duplication in the teleost fish Tetraodon nigroviridis reveals the early vertebrate proto-karyotype

Author

Jaillon, Olivier, Aury, Jean-Marc, Brunet, Frederic, Petit, Jean-Louis, Stange-Thomann, Nicole, Mauceli, Evan, Bouneau, Laurence, Fischer, Cecile, Ozouf-Costaz, Catherine, Bernot, Alain, Nicaud, Sophie, Jaffe, David, Fisher, Sheila, Lutfalla, Georges, Dossat, Carole, Segurens, Beatrice, Dasilva, Corinne, Salanoubat, Marcel, Levy, Michael, Boudet, Nathalie, Castellano, Sergi, Anthouard, Veronique, Jubin, Claire, Castelli, Vanina, Katinka, Michael, Vacherie, Benoit, Biemont, Christian, Skalli, Zineb, Cattolico, Laurence, Poulain, Julie, de Berardinis, Veronique, Cruaud, Corinne, Duprat, Simone, Brottier, Philippe, Coutanceau, Jean-Pierre, Gouzy, Jerome, Parra, Genis, Lardier, Guillaume, Chapple, Charles, McKernan, Kevin J., McEwan, Paul, Bosak, Stephanie, Kellis, Manolis, Volff, Jean-Nicolas, Guigo, Roderic, Zody, Michael C., Mesirov, Jill, Lindblad-Toh, Kerstin, Birren, Bruce, Nusbaum, Chad, Kahn, Daniel, Robinson-Rechavi, Marc, Laudet, Vincent, Schachter, Vincent, Quetier, Francis, Saurin, William, Scarpelli, Claude, Wincker, Patrick, Lander, Eric S., Weissenbach, Jean, and Roest Crollius, Hugues

Subjects
Environmental issues, Science and technology, Zoology and wildlife conservation
Abstract

Author(s): Olivier Jaillon [1]; Jean-Marc Aury [1]; Frédéric Brunet [2]; Jean-Louis Petit [1]; Nicole Stange-Thomann [3]; Evan Mauceli [3]; Laurence Bouneau [1]; Cécile Fischer [1]; Catherine Ozouf-Costaz [4]; Alain Bernot [...]

Published
2004
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35. TRIM8 is required for virus-induced IFN response in human plasmacytoid dendritic cells

Author

Maarifi, Ghizlane, primary, Smith, Nikaïa, additional, Maillet, Sarah, additional, Moncorgé, Olivier, additional, Chamontin, Célia, additional, Edouard, Joanne, additional, Sohm, Frédéric, additional, Blanchet, Fabien P., additional, Herbeuval, Jean-Philippe, additional, Lutfalla, Georges, additional, Levraud, Jean-Pierre, additional, Arhel, Nathalie J., additional, and Nisole, Sébastien, additional

Published
2019
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40. Behavior of a cloned murine interferon alpha/beta receptor expressed in homospecific or heterospecific background

Author

Uze, Gilles, Lutfalla, Georges, Bandu, Marie-Therese, and Proudhon, Dominique

Subjects
Mice -- Physiological aspects, Interferon alpha -- Physiological aspects, Interferon beta -- Physiological aspects, Cell receptors -- Genetic aspects, Science and technology
Abstract

The murine interferon alpha/beta receptor of the L1210 mouse line was cloned and characterized. Sequence analyses showed that the murine receptor is highly similar the human receptor, differing mainly in its size, being 1.2 kb longer than the human counterpart. Furthermore, the putative extracellular domains show similar organization as the human receptors. These results suggest that the murine receptor is also a cytokine receptor. Complementation studies on a receptor-deficient mutant mouse line showed that the cloned gene could complement the expression of the deficient gene, resulting in sensitivity to all alpha/beta subtypes tested.

Published
1992

42. Neutrophils use superoxide to control bacterial infection at a distance

Author

Phan, Quang Tien, Sipka, Tamara, Gonzalez, Catherine, Levraud, Jean-Pierre, Lutfalla, Georges, Nguyen-Chi, Mai, Dynamique des interactions membranaires normales et pathologiques (DIMNP), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Macrophages et Développement de l’Immunité, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Cellules Souches, Plasticité Cellulaire, Médecine Régénératrice et Immunothérapies (IRMB), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), This work was supported by a grant from the European Community’s Seventh Framework Program (FP7- PEOPLE-2011-ITN) under the Marie-Curie Initial Training Network FishForPharma [grant agreement no. PITN-GA-2011-289209] and by a grant from the European Community’s H2020 Program [Marie-Curie Innovative Training Network ImageInLife: Grant Agreement n° 721537]. QTP has been supported first by FishForPharma, then by Fondation de la Recherche Médicale (FDT20150532259). TS is supported by ImageInLife. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., European Project: 289209,EC:FP7:PEOPLE,FP7-PEOPLE-2011-ITN,FISHFORPHARMA(2012), European Project: 721537,ImageInLife(2017), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Montpellier 1 (UM1), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)

Subjects
lcsh:Immunologic diseases. Allergy, Bacterial Diseases, Embryology, Life Cycles, Neutrophils, Imaging Techniques, Immune Cells, Immunology, Research and Analysis Methods, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, White Blood Cells, Larvae, Fluorescence Microscopy, Animal Cells, Superoxides, Fluorescence Imaging, Medicine and Health Sciences, Escherichia coli, Animals, lcsh:QH301-705.5, Escherichia coli Infections, Zebrafish, Microscopy, Blood Cells, Macrophages, Embryos, Biology and Life Sciences, Light Microscopy, Cell Biology, Infectious Diseases, lcsh:Biology (General), Cellular Types, lcsh:RC581-607, Notochords, Research Article, Developmental Biology
Abstract

Understanding the roles of neutrophils and macrophages in fighting bacterial infections is a critical issue in human pathologies. Although phagocytic killing has been extensively studied, little is known about how bacteria are eliminated extracellularly in live vertebrates. We have recently developed an infection model in the zebrafish embryo in which leukocytes cannot reach the injected bacteria. When Escherichia coli bacteria are injected within the notochord, both neutrophils and macrophages are massively recruited during several days, but do not infiltrate the infected tissue presumably because of its tough collagen sheath. Nevertheless, the bacteria are killed during the first 24 hours, and we report here that neutrophils, but not macrophages are involved in the control of the infection. Using genetic and chemical approaches, we show that even in absence of phagocytosis, the bactericidal action relies on NADPH oxidase-dependent production of superoxide in neutrophils. We thus reveal a host effector mechanism mediated by neutrophils that eliminates bacteria that cannot be reached by phagocytes and that is independent of macrophages, NO synthase or myeloperoxidase., Author summary Deciphering the defence mechanisms of leukocytes remains a challenge for public health. Although phagocytic killing has been extensively studied, little is known about how bacteria are eliminated extracellularly in live vertebrates. Herein we use the notochord infection model in the zebrafish embryo to describe how leukocytes eliminate distant bacteria that are inaccessible for phagocytosis. In this context neutrophils but not macrophages are instrumental for bacterial clearance and larva survival. We then found that neutrophil bactericidal action relies on the NADPH oxidase dependent production of superoxide and is independent of NO synthase or myeloperoxidase.

Published
2018
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43. Comparative genomic analysis reveals independent expansion of a lineage-specific gene family in vertebrates: The class II cytokine receptors and their ligands in mammals and fish

Author

Mogensen Knud, Jaillon Olivier, Stange-thomann Nicole, Crollius Hugues, Lutfalla Georges, and Monneron Danièle

Subjects
Biotechnology, TP248.13-248.65, Genetics, QH426-470
Abstract

Abstract Background The high degree of sequence conservation between coding regions in fish and mammals can be exploited to identify genes in mammalian genomes by comparison with the sequence of similar genes in fish. Conversely, experimentally characterized mammalian genes may be used to annotate fish genomes. However, gene families that escape this principle include the rapidly diverging cytokines that regulate the immune system, and their receptors. A classic example is the class II helical cytokines (HCII) including type I, type II and lambda interferons, IL10 related cytokines (IL10, IL19, IL20, IL22, IL24 and IL26) and their receptors (HCRII). Despite the report of a near complete pufferfish (Takifugu rubripes) genome sequence, these genes remain undescribed in fish. Results We have used an original strategy based both on conserved amino acid sequence and gene structure to identify HCII and HCRII in the genome of another pufferfish, Tetraodon nigroviridis that is amenable to laboratory experiments. The 15 genes that were identified are highly divergent and include a single interferon molecule, three IL10 related cytokines and their potential receptors together with two Tissue Factor (TF). Some of these genes form tandem clusters on the Tetraodon genome. Their expression pattern was determined in different tissues. Most importantly, Tetraodon interferon was identified and we show that the recombinant protein can induce antiviral MX gene expression in Tetraodon primary kidney cells. Similar results were obtained in Zebrafish which has 7 MX genes. Conclusion We propose a scheme for the evolution of HCII and their receptors during the radiation of bony vertebrates and suggest that the diversification that played an important role in the fine-tuning of the ancestral mechanism for host defense against infections probably followed different pathways in amniotes and fish.

Published
2003
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44. Expression of fluorescent proteins in Branchiostoma lanceolatum by mRNA injection into unfertilized oocytes

Author

Hirsinger, Estelle, Carvalho, João Emanuel, Chevalier, Christine, Lutfalla, Georges, Nicolas, Jean-François, Peyriéras, Nadine, Schubert, Michael, Département de Biologie du Développement et Cellules souches - Department of Developmental and Stem Cell Biology, Institut Pasteur [Paris] (IP), Laboratoire de Biologie du Développement de Villefranche sur mer (LBDV), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de la Mer de Villefranche (IMEV), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Dynamique des interactions membranaires normales et pathologiques (DIMNP), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), BioEmergences, Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Centre National de la Recherche Scientifique (CNRS), This work was supported by funds from ANR (ANR-09-BLAN-0262-02 and ANR-11-JSV2-002-01) to Michael Schubert, by the European Union FP6 grant 'Embryomics' and by the ANR grant 'ANR-10-BLAN-121801 Dev-Process' to Jean-François Nicolas and Nadine Peyriéras. João Emanuel Carvalho is financed by a FCT doctoral fellowship (SFRH/BD/86878/2012)., ANR-09-BLAN-0262,EvolAx,Origine et diversification des mécanismes de régionalisation de la blastula chez les cordés(2009), Bases Génétiques, Moléculaires et Cellulaires du Développement (BGMCD), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Observatoire océanologique de Villefranche-sur-mer (OOVM), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), BioEmergences, CNRS UPS3674, Institut des Systèmes Complexes - Paris Ile-de-France (ISC-PIF), École normale supérieure - Cachan (ENS Cachan)-Université Panthéon-Sorbonne (UP1)-Université Paris-Sud - Paris 11 (UP11)-Université Pierre et Marie Curie - Paris 6 (UPMC)-École polytechnique (X)-Institut Curie-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Montpellier 1 (UM1), LUTFALLA, Georges, and Blanc - Origine et diversification des mécanismes de régionalisation de la blastula chez les cordés - - EvolAx2009 - ANR-09-BLAN-0262 - Blanc - VALID

Subjects
Male, Amphioxus, animal structures, [SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, cephalochordate, Microinjections, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, gene expression vectors, Green Fluorescent Proteins, [SDV.BDD.EO] Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, microinjection protocol, Luminescent Proteins, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Oocytes, Animals, Female, RNA, Messenger, in vivo imaging, Issue 95, model organism, ComputingMilieux_MISCELLANEOUS, Lancelets, Developmental Biology
Abstract

International audience; We report here a robust and efficient protocol for the expression of fluorescent proteins after mRNA injection into unfertilized oocytes of the cephalochordate amphioxus, Branchiostoma lanceolatum. We use constructs for membrane and nuclear targeted mCherry and eGFP that have been modified to accommodate amphioxus codon usage and Kozak consensus sequences. We describe the type of injection needles to be used, the immobilization protocol for the unfertilized oocytes, and the overall injection set-up. This technique generates fluorescently labeled embryos, in which the dynamics of cell behaviors during early development can be analyzed using the latest in vivo imaging strategies. The development of a microinjection technique in this amphioxus species will allow live imaging analyses of cell behaviors in the embryo as well as gene-specific manipulations, including gene overexpression and knockdown. Altogether, this protocol will further consolidate the basal chordate amphioxus as an animal model for addressing questions related to the mechanisms of embryonic development and, more importantly, to their evolution.

Published
2015

46. TNF signaling and macrophages govern fin regeneration in zebrafish larvae

Author

Nguyen-Chi, Mai, Laplace-Builhé, Béryl, Travnickova, Jana, Luz-Crawford, Patricia, Tejedor, Gautier, Lutfalla, Georges, Kissa, Karima, Jorgensen, Christian, Djouad, Farida, Cellules Souches, Plasticité Cellulaire, Médecine Régénératrice et Immunothérapies (IRMB), Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Dynamique des interactions membranaires normales et pathologiques (DIMNP), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Montpellier 1 (UM1), Universidad de los Andes [Santiago] (UANDES), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), This work was supported by Inserm and grants from ‘Région Languedoc-Roussillon, Chercheur d’Avenir’ no. DGA3/DESR 2012/Q209, from the French National Research Agency ‘Zebraflam’ no. ANR-10-MIDI-009 and from the European Community’s Seventh Framework Program (FP7- PEOPLE-2011-ITN) under the Marie-Curie Initial Training Network FishForPharma (Grant Agreement No. PITN-GA-2011-289209). We thank the MRI facility for their assistance, J-P. Levraud (Institut Pasteur, France) for trans-shipping Tg(mpeg:Gal4, UAS:NTR-mCherry) line, A. Kawakami (Tokyo Institute of Technology, Japan) for junbl probe, M. Bagnat for tg(rcn3:gal4/UAS:dsRed) line and E. Lelièvre (Université de Montpellier, Montpellier) for providing support and advice during parabiosis experiments., ANR-10-MIDI-0009,ZebraFlam,Signaux et cellules de la réponse inflammatoire: suivi en temps réel chez un vertébré entier, le danio zébré(2010), European Project: 289209,EC:FP7:PEOPLE,FP7-PEOPLE-2011-ITN,FISHFORPHARMA(2012), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), LUTFALLA, Georges, MECANISMES INTEGRES DE L'INFLAMMATION - Signaux et cellules de la réponse inflammatoire: suivi en temps réel chez un vertébré entier, le danio zébré - - ZebraFlam2010 - ANR-10-MIDI-0009 - MI2 - VALID, and Training Network on Zebrafish Infection Models for Pharmaceutical Screens - FISHFORPHARMA - - EC:FP7:PEOPLE2012-01-01 - 2015-12-31 - 289209 - VALID

Subjects
animal structures, Tumor Necrosis Factor-alpha, Macrophage, Macrophages, fungi, Correction, Parabiosis, Extremities, Zebrafish Proteins, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Animals, Genetically Modified, Receptors, Tumor Necrosis Factor, Type I, Larva, Animals, Regeneration, Original Article, Blastema, Fin regeneration, Amputation, Natural regeneration, [SDV.IMM.II] Life Sciences [q-bio]/Immunology/Innate immunity, Zebrafish, Signal Transduction
Abstract

International audience; Macrophages are essential for appendage regeneration after amputation in regenerative species. The molecular mechanisms through which macrophages orchestrate blastema formation and regeneration are still unclear. Here, we use the genetically tractable and transparent zebrafish larvae to study the functions of polarized macrophage subsets during caudal fin regeneration. After caudal fin amputation, we show an early and transient accumulation of pro-inflammatory macrophages concomitant with the accumulation of non-inflammatory macrophages which, in contrast to pro-inflammatory macrophages, remain associated to the fin until the end of the regeneration. Chemical and genetic depletion of macrophages suggested that early recruited macrophages that express TNFα are critical for blastema formation. Combining parabiosis and morpholino knockdown strategies, we show that TNFα/TNFR1 signaling pathway is required for the fin regeneration. Our study reveals that TNFR1 has a necessary and direct role in blastema cell activation suggesting that macrophage subset balance provides the accurate TNFα signal to prime regeneration in zebrafish.

Published
2017

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