Abstract

Hepcidin, an antimicrobial peptide produced by the liver, also controls the iron balance and regeneration in vertebrates. Two types of hepcidin (Hamp1 and Hamp2) have been found in the bodies of black rockfish (Sebastes schlegelii). The full-length cDNA of hepcidin was cloned to enable a study of the antibacterial roles of these two hepcidins (Hamp) in black rockfish. The antimicrobial function of recombinant hepcidins was tested both in vitro and in vivo by the synthesis in Escherichia coli of recombinant hepcidin (approximately 11 kDa) from black rockfish. The recombinant hepcidins inhibited the growth of two bacterial species, Streptococcus iniae FP5228 and Pseudomonas aeruginosa, at various concentrations, in vitro after 6 h post-incubation, respectively. During infection, the production of ferroportin was reduced, suggesting the preservation of iron to prevent microbial proliferation. In vivo administration of Hamp1, but not Hamp2, synthetic peptides induced a substantial reduction in the expression of ferroportin, suggesting that in black rockfish with two forms of hepcidin, ferroportin production is regulated by the iron-regulator Hamp1, and not by the dedicated antimicrobial Hamp2. The findings of this study suggest the various antimicrobial roles of these two types of hepcidin.

Highlights

  • Innate immunity is the first line of defense against infections for fish [1]

  • We described olive flounder hepcidin, which exhibited a wide range of antibacterial activity against Edwardsiella tarda, Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, and Aeromonas hydrophila [24]

  • As a regulator of iron metabolism, hepcidin synthesis is regulated by multiple factors via multiple mechanisms, caused by high iron levels and infection/inflammation, and suppressed by low iron, anemia, and hypoxia

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Summary

Introduction

Innate immunity is the first line of defense against infections for fish [1]. One of the major components of innate immunity is a fish’s rapid physiological reaction to injury or infection lasting 1–2 days. There is currently increased interest in understanding the function of hepcidin-like peptides in the natural immunity of fish [8] This small peptide has demonstrated antimicrobial activity against bacterial, viral, fungal, and parasitic infections in different species of fish [9,10,11,12]. Increased transcript levels for hepcidin have been observed in the spleen, liver, and kidney tissues of infected Lates calcarifer during parasitic infections with Cryptocaryon irritans [21]. These results indicate that hepcidins have an inductive response to antigenic stimulus and differing temporal transmission, indicating their strong capacity for customizing a unique inborn immunity [22]. Two hepcidin transcripts of an economically important species of melanops, black rockfish (Sebastes schlegelii), which constitutes a significant proportion of global aquacultural production, have been identified in the present study

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