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 inPloS one Vol. 10; no. 10; p. e0138949
Main Authors Walton, Eric M, Cronan, Mark R, Beerman, Rebecca W, Tobin, David M
Format Journal Article
LanguageEnglish
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.
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
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  givenname: Eric M
  surname: Walton
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  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
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  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
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  givenname: David M
  surname: Tobin
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  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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ContentType Journal Article
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.
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– notice: 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.
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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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SSID ssj0053866
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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
URI https://www.ncbi.nlm.nih.gov/pubmed/26445458
https://www.proquest.com/docview/1720162885
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https://pubmed.ncbi.nlm.nih.gov/PMC4596833
https://doaj.org/article/3ed02d6d5e7e4e95b475a3b7ac432852
http://dx.doi.org/10.1371/journal.pone.0138949
Volume 10
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