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Molecular Analysis and Sex-specific Response of the Hepcidin Gene in Yellow Perch (Perca Flavescens) Following Lipopolysaccharide Challenge

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Abstract

Hepcidin antimicrobial peptide (hamp) is active in teleosts against invading pathogens and plays important roles in the stress and immune responses of finfish. The response of hamp gene was studied in yellow perch (yp) (Perca flavescens) challenged with lipopolysaccharides to understand if this immunity response is sex-specifically different. The cloned hamp gene consists of an open-reading frame of 273 bp and encodes a deduced protein of 90 amino acids (a.a.), which includes a signal peptide of 24 a.a., a pro-domain of 40 a.a. and a mature peptide of 26 a.a. Yp hamp involves 8 cysteine residues with 4 disulfide bonds, and a protein with an internal alpha helix flanked with C- and N-terminal random coils was modeling predicted. RT-qPCR was used to analyze the relative abundances (RAs) of hamp mRNA in the livers of juvenile female and male yellow perch challenged with lipopolysaccharide. The expression levels of hamp were significantly elevated by 3 h (RA = 7.3) and then peaked by 6 h (RA = 29.4) post-treatment in females but the peak was delayed to 12 h (RA = 65.4) post-treatment in males. The peak mRNA level of challenged males was shown 7.6-fold higher than females. The post-treatment responses in both genders decreased to their lowest levels by 24 h and 48 h. Overall, female perch had an earlier but less-sensitive response to the lipopolysaccharide challenge than male.

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Data Availability

Raw data were generated at USDA-ARS in School of Freshwater Sciences of University of Wisconsin in Milwaukee. Derived data supporting the finding of this study are available from the corresponding author, Dr. Yuehchiang Han, on request.

Abbreviations

Hamp:

Hepcidin antimicrobial peptide

Yp:

Yellow perch

A.a.:

Amino acids

RA:

Relative abundance

AMPs:

Antimicrobial peptides

LPS:

Lipopolysaccharide

RT-qPCR:

Quantitative real-time poly chain reaction

Eflα:

Elongation factor 1 alpha

UPGMA:

Unweighted pair group method with arithmetic means

WAG:

Whelan and Goldman

ANOVA:

Analysis of variance

REML:

Restricted maximum likelihood

CDS:

Coding sequence

ORF:

Open-reading frame

MSA:

Multiple sequence alignment

SOCS:

Suppressors of cytokine signaling

EDC:

Endocrine disruption chemical

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Acknowledgements

This work was funded by USDA-ARS in-house project numbers 5090-31320-004-00D, 5090-31320-005-00D and 8001-88888-002-00D, and in part by an appointment of Y.-C.H. to the Research Participation Program at the Oak Ridge Institute of Science and Education (ORISE) through an interagency agreement between the US Department of Energy and the USDA. ORISE is managed by Oak Ridge Associated Universities (ORAU) under DOE contract DE-AC05-06OR23100. Partial support was granted from the funding (contract #019585F) awarded to Dong-Fang Deng by North Center Regional Aquaculture Center, USDA-NIFA. Xing Lu was supported by the China Scholarship Council as a visiting scientist (No. 201803260002). The views contained in this document are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of the US Government. Mention of trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA and does not imply its approval to the exclusion of other products that may be suitable. This manuscript is submitted for publication with the understanding that the US Government is authorized to reproduce and distribute reprints for governmental purposes. The USDA is an equal opportunity employee.

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Authors and Affiliations

  1. School of Freshwater Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI, 53204, USA

    Xing Lu, Ying Xiang & Dong-Fang Deng

  2. Oak Ridge Institute for Science and Education-Oak Ridge Associated Universities-US Department of Agriculture-Agricultural Research Service-School of Freshwater Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI, 53204, USA

    Yueh-Chiang Han

  3. US Department of Agriculture-Agricultural Research Service-School of Freshwater Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI, 53204, USA

    Brian S. Shepherd

  4. US Department of Agriculture-Agricultural Research Service/Northeast Area Statistics Group, Beltsville, MD, USA

    Bryan T. Vinyard

  5. Key Laboratory of Freshwater Biodiversity Conservation and Utilization of Ministry of Agriculture, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, 430223, Hubei, China

    Xing Lu

  6. School of Basic Medical Science, Wuhan University, Wuhan, 430071, Hubei, China

    Ying Xiang

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Contributions

All authors contributed to experimental design and review and approval of submitted manuscript. Study conceptualization, D.F.D., X.L., & B.S.S. Cloning & sequencing, X.L. & B.S.S. RT-qPCR analysis, Y.-C.H., X.L., & Y.X. Bioinformatics analyses & structural modeling, Y.-C.H. & X.L. Statistical analysis, B.T.V. & Y.-C.H. Writing & editing manuscript, Y.-C.H., X.L., D.F.D., & B.S.S.

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Correspondence to Yueh-Chiang Han.

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No animals were used for this specific study, but rather, tissue samples from a study that was previously approved by the University of Wisconsin at Milwaukee Animal Care and Use Committee.

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Lu, X., Han, YC., Shepherd, B.S. et al. Molecular Analysis and Sex-specific Response of the Hepcidin Gene in Yellow Perch (Perca Flavescens) Following Lipopolysaccharide Challenge. Probiotics & Antimicro. Prot. 15, 215–225 (2023). https://doi.org/10.1007/s12602-022-10024-8

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