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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| Format: | Article |
| Language: | English |
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MDPI AG
2020-08-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/18/9/439 |
| _version_ | 1797555871621840896 |
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| author | Louis Benoist Baptiste Houyvet Joël Henry Erwan Corre Bruno Zanuttini Céline Zatylny-Gaudin |
| author_facet | Louis Benoist Baptiste Houyvet Joël Henry Erwan Corre Bruno Zanuttini Céline Zatylny-Gaudin |
| author_sort | Louis Benoist |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-10T16:54:39Z |
| format | Article |
| id | doaj.art-1d115db9f7e243d3996a2edfb546b41c |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-10T16:54:39Z |
| publishDate | 2020-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-1d115db9f7e243d3996a2edfb546b41c2023-11-20T11:11:23ZengMDPI AGMarine Drugs1660-33972020-08-0118943910.3390/md18090439In-Depth In Silico Search for Cuttlefish (<i>Sepia officinalis)</i> Antimicrobial Peptides Following Bacterial Challenge of HaemocytesLouis Benoist0Baptiste Houyvet1Joël Henry2Erwan Corre3Bruno Zanuttini4Céline Zatylny-Gaudin5Normandy University, Unicaen, CNRS, BOREA, 14000 CAEN, FranceNormandy University, Unicaen, CNRS, BOREA, 14000 CAEN, FranceNormandy University, Unicaen, CNRS, BOREA, 14000 CAEN, FrancePlateforme ABiMS, Station Biologique de Roscoff (CNRS-Sorbonne Université), 29688 Roscoff, FranceNormandy University, Unicaen, Ensicaen, CNRS, GREYC, 14000 Caen, FranceNormandy 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 |
| 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 |
| title | 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_short | 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 |
| 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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