Document Detail


Functional characterization of full-length TLR3, IRAK-4, and TRAF6 in zebrafish (Danio rerio).
MedLine Citation:
PMID:  15829296     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recently, the zebrafish, Danio rerio, has been recognized as a useful model for infectious disease and immunity. The Toll-like receptor (TLR) family is an evolutionarily conserved component of the innate immune system that responds to specific pathogen-associated molecular patterns (PAMPs) during an infection. This study reports the identification and characterization of a full-length orthologue of mammalian TLR3, and the key TLR pathway signaling molecules IRAK-4 and TRAF6 in the zebrafish. Sequence analysis of zebrafish TLR3 (zfTLR3), IRAK-4 (zfIRAK-4), and TRAF6 (zfTRAF6) revealed conserved domains shared with insect and mammalian genes. Quantitative real-time PCR showed that all three genes are expressed in a variety of adult tissues and during embryonic development. In in situ hybridization, we showed that zfTLR3, zfIRAK-4, and zfTRAF6 are present in distinct regions of the developing brain at 22hpf and that zfTRAF6 was observed in the developing medial neural tube. Overexpression of zfIRAK-4, zfTRAF6, or a mutant zfTLR3 construct was able to stimulate NF-kappaB activation in ZFL cells as measured by a cotransfected NF-kappaB-luciferase reporter plasmid. Messenger RNA expression profiles of each gene in zebrafish embryos and adults were examined by quantitative real-time PCR following infection with snakehead rhabdovirus (SHRV) or Edwardsiella tarda. Following exposure to SHRV, only zfTLR3 and zfTRAF6 mRNA transcripts were upregulated. Interestingly, exposure of fish to E. tarda resulted in an unexpected increase in mRNA expression of zfTLR3, as well as the anticipated upregulation of zfIRAK-4 and zfTRAF6 mRNA transcripts. These results demonstrate that zebrafish possess conserved TLR-signaling pathways, further emphasizing the utility of the zebrafish as a model for vertebrate immunology.
Authors:
Peter E Phelan; Mark T Mellon; Carol H Kim
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.     Date:  2005-01-05
Journal Detail:
Title:  Molecular immunology     Volume:  42     ISSN:  0161-5890     ISO Abbreviation:  Mol. Immunol.     Publication Date:  2005 May 
Date Detail:
Created Date:  2005-04-14     Completed Date:  2005-07-12     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7905289     Medline TA:  Mol Immunol     Country:  England    
Other Details:
Languages:  eng     Pagination:  1057-71     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Microbiology, and Molecular Biology, 5735 Hitchner Hall, University of Maine, Orono, ME 04469, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Cells, Cultured
Cloning, Molecular
Edwardsiella tarda / growth & development,  isolation & purification
Embryo, Nonmammalian
Enterobacteriaceae Infections / pathology
Gene Expression Regulation, Developmental
Genes, Reporter
In Situ Hybridization
Interleukin-1 Receptor-Associated Kinases
Liver / cytology
Luciferases
Membrane Glycoproteins / chemistry,  genetics,  metabolism*
Molecular Sequence Data
Mutation
NF-kappa B / metabolism
Phosphotransferases (Alcohol Group Acceptor) / chemistry,  genetics,  metabolism
Protein Structure, Tertiary
RNA, Messenger / genetics,  metabolism
Receptors, Cell Surface / chemistry,  genetics,  metabolism*
Reverse Transcriptase Polymerase Chain Reaction
Rhabdoviridae Infections / pathology
Sequence Analysis, Protein
Sequence Homology, Amino Acid
TNF Receptor-Associated Factor 6 / metabolism*
Toll-Like Receptor 3
Toll-Like Receptors
Zebrafish / embryology,  genetics,  metabolism*
Zebrafish Proteins / chemistry,  genetics,  metabolism*
Grant Support
ID/Acronym/Agency:
R15 AI49237-01/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Membrane Glycoproteins; 0/NF-kappa B; 0/RNA, Messenger; 0/Receptors, Cell Surface; 0/TNF Receptor-Associated Factor 6; 0/Toll-Like Receptor 3; 0/Toll-Like Receptors; 0/Zebrafish Proteins; EC 1.13.12.-/Luciferases; EC 2.7.1.-/IRAK-4 protein, zebrafish; EC 2.7.1.-/Phosphotransferases (Alcohol Group Acceptor); EC 2.7.11.1/Interleukin-1 Receptor-Associated Kinases

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