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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 37, No. 6, 2016, pp. 349-357
Bioline Code: zr16051
Full paper language: English
Document type: Report
Document available free of charge

Zoological Research, Vol. 37, No. 6, 2016, pp. 349-357

 en Molecular characterization and functional analysis of a piscidin gene in large yellow croaker ( Larimichthys crocea check for this species in other resources )
YANG, Jing; LU, Xin-Jiang; CHAI, Fang-Chao & CHEN, Jiong

Abstract

The piscidin family, which includes potent antimicrobial peptides with broad-spectrum activity, plays an important role in the innate immune system of fish. In this study, we cloned piscidin-5-like type 3 (Lcpis5lt3) in large yellow croaker ( Larimichthys crocea check for this species in other resources ). Multiple alignments with other known piscidins revealed amino acid conservation throughout the fish, especially at the signal peptide (22 amino acids). The phylogenetic tree confirmed that Lcpis5lt3 and large yellow croaker piscidin-5-like proteins were grouped together to form a branch. Quantitative real-time PCR revealed that Lcpis5lt3 was expressed in a wide range of tissues, including the brain, muscle, gill, head kidney, intestine, kidney, liver, and spleen. The highest mRNA expression level of Lcpis5lt3 was found in the spleen. After Vibrio alginolyticus check for this species in other resources infection, mRNA expression was rapidly upregulated in the liver, head kidney, gill, kidney, and intestine at 4, 8, 12, and 24 h post infection (hpi), whereas there were no significant changes in the spleen. The antimicrobial spectrum showed that the synthetic mature peptide of Lcpis5lt3 exhibited different activity in vitro against various bacteria, such as Aeromonas hydrophila check for this species in other resources , Vibrio anguillarum check for this species in other resources , Vibrio alginolyticus, Vibrio parahaemolyticus check for this species in other resources , Staphylococcus aureus check for this species in other resources , and Listeria monocytogenes check for this species in other resources . In addition, survival rates from the in vivo assay indicated that the synthetic peptide of Lcpis5lt3 increased the survival rate of large yellow croaker after V. alginolyticus challenge, resulting in a decline in bacterial burden and mRNA expression levels of interleukin-1β, interleukin-10, and tumor necrosis factor-α. These data suggest that Lcpis5lt3 plays an important role in innate immunity in large yellow croaker and might represent a potential therapeutic agent against pathogen invasion.

Keywords
Antimicrobial activity; Large yellow croaker; Piscidin; Survival rate; Vibrio alginolyticus

 
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