In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of Haemocytes
Cuttlefish (<i>Sepia officinalis</i>) haemocytes are potential sources of antimicrobial peptides (AMPs). To study the immune response to <i>Vibrio splendidus</i> and identify new AMPs, an original approach was developed based on a differential transcriptomic study and an in-d...
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doaj-1d115db9f7e243d3996a2edfb546b41c2020-11-25T03:40:08ZengMDPI AGMarine Drugs1660-33972020-08-011843943910.3390/md18090439In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of HaemocytesLouis Benoist0Baptiste Houyvet1Joël Henry2Erwan Corre3Bruno Zanuttini4Céline Zatylny-Gaudin5Normandie University, Unicaen, CNRS, BOREA, 14000 CAEN, FranceNormandie University, Unicaen, CNRS, BOREA, 14000 CAEN, FranceNormandie University, Unicaen, CNRS, BOREA, 14000 CAEN, FrancePlateforme ABiMS, Station Biologique de Roscoff (CNRS-Sorbonne Université), 29688 Roscoff, FranceNormandie University, Unicaen, Ensicaen, CNRS, GREYC, 14000 Caen, FranceNormandie University, Unicaen, CNRS, BOREA, 14000 CAEN, FranceCuttlefish (<i>Sepia officinalis</i>) haemocytes are potential sources of antimicrobial peptides (AMPs). To study the immune response to <i>Vibrio splendidus</i> 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 <i>V</i>. <i>splendidus</i> or not. A first analysis of the annotated transcripts revealed the presence of Toll/NF-κB pathway members, including newly identified factors such as <i>So</i>-TLR-h, <i>So</i>-IKK-h and <i>So</i>-Rel/NF-κB-h. Out of the eight Toll/NF-κB pathway members, seven were found up-regulated following <i>V</i>. <i>splendidus</i> 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 <i>Vibrio</i> 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.https://www.mdpi.com/1660-3397/18/9/439antimicrobial peptidehaemocyte<i>Sepia officinalis</i>molluscchallengein vitro |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Louis Benoist Baptiste Houyvet Joël Henry Erwan Corre Bruno Zanuttini Céline Zatylny-Gaudin |
spellingShingle |
Louis Benoist Baptiste Houyvet Joël Henry Erwan Corre Bruno Zanuttini Céline Zatylny-Gaudin In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of Haemocytes Marine Drugs antimicrobial peptide haemocyte <i>Sepia officinalis</i> mollusc challenge in vitro |
author_facet |
Louis Benoist Baptiste Houyvet Joël Henry Erwan Corre Bruno Zanuttini Céline Zatylny-Gaudin |
author_sort |
Louis Benoist |
title |
In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of Haemocytes |
title_short |
In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of Haemocytes |
title_full |
In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of Haemocytes |
title_fullStr |
In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of Haemocytes |
title_full_unstemmed |
In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of Haemocytes |
title_sort |
in-depth in silico search for cuttlefish (<i>sepia officinalis)</i> antimicrobial peptides following bacterial challenge of haemocytes |
publisher |
MDPI AG |
series |
Marine Drugs |
issn |
1660-3397 |
publishDate |
2020-08-01 |
description |
Cuttlefish (<i>Sepia officinalis</i>) haemocytes are potential sources of antimicrobial peptides (AMPs). To study the immune response to <i>Vibrio splendidus</i> 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 <i>V</i>. <i>splendidus</i> or not. A first analysis of the annotated transcripts revealed the presence of Toll/NF-κB pathway members, including newly identified factors such as <i>So</i>-TLR-h, <i>So</i>-IKK-h and <i>So</i>-Rel/NF-κB-h. Out of the eight Toll/NF-κB pathway members, seven were found up-regulated following <i>V</i>. <i>splendidus</i> 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 <i>Vibrio</i> 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. |
topic |
antimicrobial peptide haemocyte <i>Sepia officinalis</i> mollusc challenge in vitro |
url |
https://www.mdpi.com/1660-3397/18/9/439 |
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