PUBLICATION
Immune response and inhibition of bacterial growth by electrotransfer of plasmid DNA containing the antimicrobial peptide, epinecidin-1, into zebrafish muscle
- Authors
- Lin, S.B., Fan, T.W., Wu, J.L., Hui, C.F., and Chen, J.Y.
- ID
- ZDB-PUB-090407-13
- Date
- 2009
- Source
- Fish & shellfish immunology 26(3): 451-458 (Journal)
- Registered Authors
- Chen, Jyh-Yih, Wu, Jen-Leih
- Keywords
- Antimicrobial peptide, Zebrafish, Epinecidin-1, Electrotransfer
- MeSH Terms
-
- Animals
- Antimicrobial Cationic Peptides/genetics*
- Antimicrobial Cationic Peptides/immunology*
- Colony Count, Microbial
- Cytokines/genetics
- DNA/genetics
- DNA/immunology*
- Electroporation
- Fish Proteins/genetics*
- Fish Proteins/immunology*
- Gene Expression Regulation
- Immunohistochemistry
- Muscle, Skeletal/immunology*
- Muscle, Skeletal/microbiology*
- Plasmids/genetics
- Promoter Regions, Genetic
- Reverse Transcriptase Polymerase Chain Reaction
- Vibrio vulnificus/growth & development
- Vibrio vulnificus/physiology*
- Zebrafish/physiology*
- PubMed
- 19340938 Full text @ Fish Shellfish Immunol.
Citation
Lin, S.B., Fan, T.W., Wu, J.L., Hui, C.F., and Chen, J.Y. (2009) Immune response and inhibition of bacterial growth by electrotransfer of plasmid DNA containing the antimicrobial peptide, epinecidin-1, into zebrafish muscle. Fish & shellfish immunology. 26(3):451-458.
Abstract
Bacterial infections represent serious diseases in aquaculture, rapidly leading to fish death by septicemia. We investigated whether the electrotransfer of green fluorescent protein gene fusion epinecidin-1 (CMV-gfp-epi) DNA into zebrafish muscle could regulate the fish immune response and inhibit bacterial growth. Electroporation parameters such as the number of pulses, voltage, and amount of plasmid DNA were analyzed, and results demonstrated the greatest mRNA expression level of gfp-epi relative to beta-actin was obtained with a pulse number of 4, a voltage strength of 100 V/cm, a concentration of DNA of 90 microg/fish, and electroporation for 96 h. In addition, the cytomegalovirus (CMV) promoter exhibited higher activity compared to the mylz promoter in muscle for electrotransfer in zebrafish. GFP fluorescence and gfp-epi mRNA expression in tissues after electroporation were also studied by a polymerase chain reaction, immunohistochemistry, and fluorescence microscopy. gfp-epi expression was significantly correlated with decreased bacterial numbers and immune-related gene expression. These data demonstrate that electroporation of epinecidin-1 might have provoked an inflammatory response that accounts for the improvement in bacterial clearance.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping