In-Depth In Silico Search for Cuttlefish (Sepia officinalis) Antimicrobial Peptides Following Bacterial Challenge of Haemocytes

Laboratoire de Biologie des Organismes et Ecosystèmes Aquatiques (BOREA) Université de Caen-Normandie, MNHN, SU, UA, CNRS, IRD, Esplanade de la Paix, CEDEX, 14032 Caen, France

SATMAR, Société ATlantique de MARiculture, Research and Development Department, 50760 Gatteville, France

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Plateforme ABiMS, Station Biologique de Roscoff (CNRS-Sorbonne Université), 29688 Roscoff, France

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Normandy University, Unicaen, Ensicaen, CNRS, GREYC, 14000 Caen, France

Author to whom correspondence should be addressed.

Mar. Drugs 2020, 18(9), 439; https://doi.org/10.3390/md18090439

Received: 23 July 2020 / Revised: 15 August 2020 / Accepted: 19 August 2020 / Published: 24 August 2020

(This article belongs to the Special Issue Marine Antibiotics 2020)

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Abstract

Cuttlefish (Sepia officinalis) haemocytes are potential sources of antimicrobial peptides (AMPs). To study the immune response to Vibrio splendidus and identify new AMPs, an original approach was developed based on a differential transcriptomic study and an in-depth in silico analysis using multiple tools. Two de novo transcriptomes were retrieved from cuttlefish haemocytes following challenge by V. splendidus or not. A first analysis of the annotated transcripts revealed the presence of Toll/NF-κB pathway members, including newly identified factors such as So-TLR-h, So-IKK-h and So-Rel/NF-κB-h. Out of the eight Toll/NF-κB pathway members, seven were found up-regulated following V. splendidus challenge. Besides, immune factors involved in the immune response were also identified and up-regulated. However, no AMP was identified based on annotation or conserved pattern searches. We therefore performed an in-depth in silico analysis of unannotated transcripts based on differential expression and sequence characteristics, using several tools available like PepTraq, a homemade software program. Finally, five AMP candidates were synthesized. Among them, NF19, AV19 and GK28 displayed antibacterial activity against Gram-negative bacteria. Each peptide had a different spectrum of activity, notably against Vibrio species. GK28—the most active peptide—was not haemolytic, whereas NF19 and AV19 were haemolytic at concentrations between 50 and 100 µM, 5 to 10 times higher than their minimum inhibitory concentration. View Full-Text

Keywords: antimicrobial peptide; haemocyte; Sepia officinalis; mollusc; challenge; in vitro; Vibrio splendidus

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MDPI and ACS Style

Benoist, L.; Houyvet, B.; Henry, J.; Corre, E.; Zanuttini, B.; Zatylny-Gaudin, C. In-Depth In Silico Search for Cuttlefish (Sepia officinalis) Antimicrobial Peptides Following Bacterial Challenge of Haemocytes. Mar. Drugs 2020, 18, 439. https://doi.org/10.3390/md18090439

AMA Style

Benoist L, Houyvet B, Henry J, Corre E, Zanuttini B, Zatylny-Gaudin C. In-Depth In Silico Search for Cuttlefish (Sepia officinalis) Antimicrobial Peptides Following Bacterial Challenge of Haemocytes. Marine Drugs. 2020; 18(9):439. https://doi.org/10.3390/md18090439

Chicago/Turabian Style

Benoist, Louis, Baptiste Houyvet, Joël Henry, Erwan Corre, Bruno Zanuttini, and Céline Zatylny-Gaudin. 2020. "In-Depth In Silico Search for Cuttlefish (Sepia officinalis) Antimicrobial Peptides Following Bacterial Challenge of Haemocytes" Marine Drugs 18, no. 9: 439. https://doi.org/10.3390/md18090439

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