The Macrophage-Specific Promoter mfap4 Allows Live, Long-Term Analysis of Macrophage Behavior during Mycobacterial Infection in Zebrafish
Transgenic labeling of innate immune cell lineages within the larval zebrafish allows for real-time, in vivo analyses of microbial pathogenesis within a vertebrate host. To date, labeling of zebrafish macrophages has been relatively limited, with the most specific expression coming from the mpeg1 pr...
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Published in | PloS one Vol. 10; no. 10; p. e0138949 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
United States
Public Library of Science
07.10.2015
Public Library of Science (PLoS) |
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Abstract | Transgenic labeling of innate immune cell lineages within the larval zebrafish allows for real-time, in vivo analyses of microbial pathogenesis within a vertebrate host. To date, labeling of zebrafish macrophages has been relatively limited, with the most specific expression coming from the mpeg1 promoter. However, mpeg1 transcription at both endogenous and transgenic loci becomes attenuated in the presence of intracellular pathogens, including Salmonella typhimurium and Mycobacterium marinum. Here, we describe mfap4 as a macrophage-specific promoter capable of producing transgenic lines in which transgene expression within larval macrophages remains stable throughout several days of infection. Additionally, we have developed a novel macrophage-specific Cre transgenic line under the control of mfap4, enabling macrophage-specific expression using existing floxed transgenic lines. These tools enrich the repertoire of transgenic lines and promoters available for studying zebrafish macrophage dynamics during infection and inflammation and add flexibility to the design of future macrophage-specific transgenic lines. |
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AbstractList | Transgenic labeling of innate immune cell lineages within the larval zebrafish allows for real-time, in vivo analyses of microbial pathogenesis within a vertebrate host. To date, labeling of zebrafish macrophages has been relatively limited, with the most specific expression coming from the mpeg1 promoter. However, mpeg1 transcription at both endogenous and transgenic loci becomes attenuated in the presence of intracellular pathogens, including Salmonella typhimurium and Mycobacterium marinum. Here, we describe mfap4 as a macrophage-specific promoter capable of producing transgenic lines in which transgene expression within larval macrophages remains stable throughout several days of infection. Additionally, we have developed a novel macrophage-specific Cre transgenic line under the control of mfap4, enabling macrophage-specific expression using existing floxed transgenic lines. These tools enrich the repertoire of transgenic lines and promoters available for studying zebrafish macrophage dynamics during infection and inflammation and add flexibility to the design of future macrophage-specific transgenic lines. Transgenic labeling of innate immune cell lineages within the larval zebrafish allows for real-time, in vivo analyses of microbial pathogenesis within a vertebrate host. To date, labeling of zebrafish macrophages has been relatively limited, with the most specific expression coming from the mpeg1 promoter. However, mpeg1 transcription at both endogenous and transgenic loci becomes attenuated in the presence of intracellular pathogens, including Salmonella typhimurium and Mycobacterium marinum . Here, we describe mfap4 as a macrophage-specific promoter capable of producing transgenic lines in which transgene expression within larval macrophages remains stable throughout several days of infection. Additionally, we have developed a novel macrophage-specific Cre transgenic line under the control of mfap4 , enabling macrophage-specific expression using existing floxed transgenic lines. These tools enrich the repertoire of transgenic lines and promoters available for studying zebrafish macrophage dynamics during infection and inflammation and add flexibility to the design of future macrophage-specific transgenic lines. |
Audience | Academic |
Author | Tobin, David M Walton, Eric M Beerman, Rebecca W Cronan, Mark R |
AuthorAffiliation | National University of Singapore, SINGAPORE 1 Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America 3 Center for Host-Microbial Interactions, Duke University Medical Center, Durham, North Carolina, United States of America 2 Department of Immunology, Duke University Medical Center, Durham, North Carolina, United States of America |
AuthorAffiliation_xml | – name: National University of Singapore, SINGAPORE – name: 1 Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America – name: 3 Center for Host-Microbial Interactions, Duke University Medical Center, Durham, North Carolina, United States of America – name: 2 Department of Immunology, Duke University Medical Center, Durham, North Carolina, United States of America |
Author_xml | – sequence: 1 givenname: Eric M surname: Walton fullname: Walton, Eric M organization: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America; Center for Host-Microbial Interactions, Duke University Medical Center, Durham, North Carolina, United States of America – sequence: 2 givenname: Mark R surname: Cronan fullname: Cronan, Mark R organization: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America; Center for Host-Microbial Interactions, Duke University Medical Center, Durham, North Carolina, United States of America – sequence: 3 givenname: Rebecca W surname: Beerman fullname: Beerman, Rebecca W organization: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America; Center for Host-Microbial Interactions, Duke University Medical Center, Durham, North Carolina, United States of America – sequence: 4 givenname: David M surname: Tobin fullname: Tobin, David M organization: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America; Department of Immunology, Duke University Medical Center, Durham, North Carolina, United States of America; Center for Host-Microbial Interactions, Duke University Medical Center, Durham, North Carolina, United States of America |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/26445458$$D View this record in MEDLINE/PubMed |
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Copyright | COPYRIGHT 2015 Public Library of Science 2015 Walton et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. 2015 Walton et al 2015 Walton et al |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Competing Interests: DMT is a PLOS ONE Editorial Board member. This does not alter the authors' adherence to PLOS ONE Editorial policies and criteria. Conceived and designed the experiments: EMW MRC RWB DMT. Performed the experiments: EMW MRC RWB. Analyzed the data: EMW MRC RWB DMT. Contributed reagents/materials/analysis tools: EMW MRC RWB. Wrote the paper: EMW MRC DMT. |
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Snippet | Transgenic labeling of innate immune cell lineages within the larval zebrafish allows for real-time, in vivo analyses of microbial pathogenesis within a... Transgenic labeling of innate immune cell lineages within the larval zebrafish allows for real-time, in vivo analyses of microbial pathogenesis within a... |
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SubjectTerms | Animals Animals, Genetically Modified - genetics Animals, Genetically Modified - microbiology Artificial chromosomes Bacterial infections Cell Lineage - genetics Cloning Danio rerio Deoxyribonucleic acid Disease Models, Animal DNA Gene expression Genetic engineering Genetics Genomes Health aspects Host-Pathogen Interactions - genetics Immune system Immunity, Innate - genetics Immunology Infection Infections Labeling Labelling Larva - genetics Larva - microbiology Macrophages Macrophages - microbiology Mammals Microorganisms Mycobacterium Infections - genetics Mycobacterium Infections - microbiology Mycobacterium marinum Mycobacterium marinum - pathogenicity Neutrophils Pathogenesis Promoter Regions, Genetic - genetics Proteins Rodents Salmonella Salmonella typhimurium - pathogenicity Transcription Transgenes - genetics Zebrafish Zebrafish - genetics Zebrafish - microbiology Zebrafish Proteins - genetics |
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Title | The Macrophage-Specific Promoter mfap4 Allows Live, Long-Term Analysis of Macrophage Behavior during Mycobacterial Infection in Zebrafish |
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