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G-protein coupled receptor 18 (GPR18) in channel catfish: Expression analysis and efficacy as immunostimulant against Aeromonas hydrophila infection

Permanent URL:
http://handle.nal.usda.gov/10113/58427
File:
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Abstract:
The objectives of this study were: 1) to determine the transcriptional profiles of G-protein coupled receptor 18 (GPR18) in channel catfish after infection with Aeromonas hydrophila compared to that in healthy catfish; 2) to determine whether over-expression of GPR18 in catfish gill cells will offer protection against infection of A. hydrophila; 3) to determine whether recombinant pcDNA-GPR18 could be used as an immunostimulant to protect channel catfish against A. hydrophila infection. Quantitative PCR revealed that the transcription levels of GPR18 in all tissues of infected catfish were significantly (P < 0.05) induced except in the intestine. When pcDNA3.2-vectored recombinant GPR18 was transfected in catfish gill cells G1B, the over-expression of pcDNA-GPR18 offered significant (P < 0.05) protection to G1B cells against A. hydrophila infection. When channel catfish were intraperitoneally injected with QCDCR adjuvant formulated pcDNA-GPR18 and challenged with a highly virulent A. hydrophila strain at 1-, 2-, 14-, and 28-days post treatment, pcDNA-GPR18 offered 50%, 100%, 57%, and 55% protection to channel catfish, respectively. Macrophages of fish treated with pcDNA-GPR18 produced significantly (P < 0.05) higher amounts of reactive oxygen species and nitric oxide than that of fish treated with pcDNA vector alone. In addition, serum lysozyme activity of catfish injected with pcDNA-GPR18 was significantly (P < 0.08) increased. Taken together, our results suggest that pcDNA-GPR18 could be used as a novel immunostimulant to provide immediate protection to channel catfish against A. hydrophila infection.
Author(s):
Julia W. Pridgeon , Phillip H. Klesius
Subject(s):
Aeromonas hydrophila , Ictalurus punctatus , bacterial infections , catfish , enzyme activity , fish diseases , gene overexpression , gills , immunization , lysozyme , macrophages , nitric oxide , polymerase chain reaction , reactive oxygen species , receptors , transcription (genetics) , transfection
Source:
Fish and Shellfish Immunology 2013 v.35
Language:
English
Year:
2013
Collection:
Journal Articles, USDA Authors, Peer-Reviewed
Rights:
Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.