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1. Influence of strains in development of francisellosis in the blue mussel Mytilus edulis during experimental challenges

Author

Hélène Bouras, Yann Quesnelle, Suzanne Trancart, Jean-Louis Blin, Manuel Savary, Céline Zatylny-Gaudin, and Maryline Houssin

Subjects
Francisella halioticida, Blue mussel, Pathogenicity, LD50, Experimental challenge, Aquaculture. Fisheries. Angling, SH1-691
Abstract

The bacterium Francisella halioticida, known to induce francisellosis in abalone and Yesso scallop, is suspected of being involved in the blue mussel mortalities observed in France. Recently, several isolates of F. halioticida were obtained from moribund mussels and categorized into two type strains, FR21 and FR22. Two other strains, AG1 and AG3, determined to belong to the genus Francisella were isolated. To determine the virulence of these isolates, juvenile and adult blue mussels were injected with bacterial solutions at high dose and monitored for 11 days. FR22c and FR22d were found to induce 80% mortality in less than seven days. The isolates AG1 and AG3 led to over 50% mortality in adult mussels but only AG1 led to significant mortality in juveniles (41%). FR22c and FR22d, the most virulent isolates, were selected to determine their respective lethal dose at 50% (LD50) in juveniles and adults. This analysis was performed with bacterial solutions ranging from 102 to 106 CFU/mussel and monitored for 30 days. The isolate FR22c was found to be the most virulent. Observed LD50 for the isolate FR22c was 4.14 ×103 CFU/juvenile and 3.45 ×103 CFU/adult and for the isolate FR22d, 1.89 ×104 CFU/juvenile and 1.52 ×104 CFU/adult. To confirm Koch’s postulate, a selection of moribund, surviving and control animals were plated on specific media. The isolates were reisolated from moribund animals but not from surviving or controls. To confirm the proliferation, a specific real-time PCR was performed. All moribund individuals were positive by PCR. The main Ct values were lower for moribund compared to surviving animals and a dose effect was observed in DNA bacterial load. This study shows that some F. halioticida isolates are able to induce francisellosis in mussels and lead to high mortality, highlighting differences in virulence among the strains.

Published
2024
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2. Involvement of GLWamide neuropeptides in polyp contraction of the adult stony coral Euphyllia ancora

Author

Shinya Shikina, Yi-Ling Chiu, Yan Zhang, Yi-ChenYao, Tai-Yu Liu, Pin-Hsuan Tsai, Céline Zatylny-Gaudin, and Ching-Fong Chang

Subjects
Medicine, Science
Abstract

Abstract The existence and function of neurons remain largely unexplored in scleractinian corals. To gain a better understanding of neuronal functions in coral physiology, this study focused on Glycine-Leucine-Tryptophan-amide family neuropeptides (GLWamides), which have been shown to induce muscle contraction and larval metamorphosis in other cnidarians. Molecular identification and functional characterization of GLWamides in the adult stony coral Euphyllia ancora were performed. We successfully elucidated the full-length cDNA of GLWamide preprohormone in E. ancora (named EaGLW preprohormone). The deduced amino acid sequence was predicted to contain six potential GLWamide peptides. Tissue distribution analysis demonstrated that transcripts of EaGLW preprohormone were mainly expressed in the mouth (including the pharynx) and tentacles of the polyps. Immunodetection with an anti-GLWamide monoclonal antibody revealed that GLWamide neurons were mainly distributed in the epidermis of the mouth region and tentacle, in agreement with the distribution patterns of the transcripts. Treatment of the isolated mouth and tentacles with synthetic GLWamide peptides induced the contraction of these isolated tissues. Treatment of polyps with synthetic GLWamide peptides induced the contraction of polyps. These results suggest that GLWamides are involved in polyp contraction (myoactivity) in adult scleractinians. Our data provide new information on the physiological function of neuropeptides in scleractinians.

Published
2020
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3. Structural and Functional Characterization of Orcokinin B-like Neuropeptides in the Cuttlefish (Sepia officinalis)

Author

Maxime Endress, Céline Zatylny-Gaudin, Jérôme Leprince, Benjamin Lefranc, Erwan Corre, Gildas Le Corguillé, Benoît Bernay, Alexandre Leduc, Jimmy Rangama, Liza Mouret, Anne-Gaëlle Lafont, Arnaud Bondon, and Joël Henry

Subjects
cephalopods, Sepia officinalis, egg-laying, neuropeptides, neurohormones, biological activity, Biology (General), QH301-705.5
Abstract

The cuttlefish (Sepia officinalis) is a Cephalopod mollusk that lives in the English Channel and breeds in coastal spawning grounds in spring. A previous work showed that the control of egg-laying is monitored by different types of regulators, among which neuropeptides play a major role. They are involved in the integration of environmental cues, and participate in the transport of oocytes in the genital tract and in the secretion of capsular products. This study addresses a family of neuropeptides recently identified and suspected to be involved in the control of the reproduction processes. Detected by mass spectrometry and immunocytochemistry in the nerve endings of the accessory sex glands of the females and ovary, these neuropeptides are also identified in the hemolymph of egg-laying females demonstrating that they also have a hormone-like role. Released in the hemolymph by the sub-esophageal mass, a region that innervates the genital tract and the neurohemal area of the vena cava, in in vitro conditions these neuropeptides modulated oocyte transport and capsular secretion. Finally, in silico analyses indicated that these neuropeptides, initially called FLGamide, had extensive structural homology with orcokinin B, which motivated their name change.

Published
2022
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4. Dietary aquaculture by-product hydrolysates: impact on the transcriptomic response of the intestinal mucosa of European seabass (Dicentrarchus labrax) fed low fish meal diets

Author

Alexandre Leduc, Céline Zatylny-Gaudin, Marie Robert, Erwan Corre, Gildas Le Corguille, Hélène Castel, Antoine Lefevre-Scelles, Vincent Fournier, Enric Gisbert, Karl B. Andree, and Joël Henry

Subjects
Hydrolysate, Fishmeal replacement, By-products, Aquaculture, Aquafeed, European seabass, Biotechnology, TP248.13-248.65, Genetics, QH426-470
Abstract

Abstract Background Aquaculture production is expected to double by 2030, and demands for aquafeeds and raw materials are expected to increase accordingly. Sustainable growth of aquaculture will require the development of highly nutritive and functional raw materials to efficiently replace fish meal. Enzymatic hydrolysis of marine and aquaculture raw materials could bring new functionalities to finished products. The aim of this study was to determine the zootechnical and transcriptomic performances of protein hydrolysates of different origins (tilapia, shrimp, and a combination of the two) in European seabass (Dicentrarchux labrax) fed a low fish meal diet (5%), for 65 days. Results Results were compared to a positive control fed with 20% of fish meal. Growth performances, anterior intestine histological organization and transcriptomic responses were monitored and analyzed. Dietary inclusion of protein hydrolysates in the low fish meal diet restored similar growth performances to those of the positive control. Inclusion of dietary shrimp hydrolysate resulted in larger villi and more goblet cells, even better than the positive control. Transcriptomic analysis of the anterior intestine showed that dietary hydrolysate inclusion restored a pattern of intestinal gene expression very close to the pattern of the positive control. However, as compared to the low fish meal diet and depending on their origin, the different hydrolysates did not modulate metabolic pathways in the same way. Dietary shrimp hydrolysate inclusion modulated more metabolic pathways related to immunity, while nutritional metabolism was more impacted by dietary tilapia hydrolysate. Interestingly, the combination of the two hydrolysates enhanced the benefits of hydrolysate inclusion in diets: more genes and metabolic pathways were regulated by the combined hydrolysates than by each hydrolysate tested independently. Conclusions Protein hydrolysates manufactured from aquaculture by-products are promising candidates to help replace fish meal in aquaculture feeds without disrupting animal metabolism and performances.

Published
2018
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5. Marine Transcriptomics Analysis for the Identification of New Antimicrobial Peptides

Author

Baptiste Houyvet, Yolande Bouchon-Navaro, Claude Bouchon, Erwan Corre, and Céline Zatylny-Gaudin

Subjects
venomous fish, lionfish, antimicrobial peptides, transcriptomic, Biology (General), QH301-705.5
Abstract

Antimicrobial peptides (AMPs) participate in the immune system to avoid infection, are present in all living organisms and can be used as drugs. Fish express numerous AMP families including defensins, cathelicidins, liver-expressed antimicrobial peptides (LEAPs), histone-derived peptides, and piscidins (a fish-specific AMP family). The present study demonstrates for the first time the occurrence of several AMPs in lionfish (Pterois volitans). Using the lionfish transcriptome, we identified four transcript sequences encoding cysteine-rich AMPs and two new transcripts encoding piscidin-like peptides. These AMPs are described for the first time in a species of the Scorpaenidae family. A functional approach on new pteroicidins was carried out to determine antimicrobial sequences and potential uses, with a view to using some of these AMPs for human health or in aquaculture.

Published
2021
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6. In-Depth In Silico Search for Cuttlefish (Sepia officinalis) Antimicrobial Peptides Following Bacterial Challenge of Haemocytes

Author

Louis Benoist, Baptiste Houyvet, Joël Henry, Erwan Corre, Bruno Zanuttini, and Céline Zatylny-Gaudin

Subjects
antimicrobial peptide, haemocyte, Sepia officinalis, mollusc, challenge, in vitro, Biology (General), QH301-705.5
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.

Published
2020
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7. How Egg Case Proteins Can Protect Cuttlefish Offspring?

Author

Valérie Cornet, Joël Henry, Didier Goux, Emilie Duval, Benoit Bernay, Gildas Le Corguillé, Erwan Corre, and Céline Zatylny-Gaudin

Subjects
Medicine, Science
Abstract

Sepia officinalis egg protection is ensured by a complex capsule produced by the female accessory genital glands and the ink bag. Our study is focused on the proteins constituting the main egg case. De novo transcriptomes from female genital glands provided essential databases for protein identification. A proteomic approach in SDS-PAGE coupled with MS unveiled a new egg case protein family: SepECPs, for Sepia officinalis Egg Case Proteins. N-glycosylation was demonstrated by PAS staining SDS-PAGE gels. These glycoproteins are mainly produced in the main nidamental glands. SepECPs share high sequence homology, especially in the signal peptide and the three cysteine-rich domains. SepECPs have a high number of cysteines, with conserved motifs involved in 3D-structure. SDS-PAGE showed that SepECPs could form dimers; this result was confirmed by TEM observations, which also revealed a protein network. This network is similar to the capsule network, and it associates these structural proteins with polysaccharides, melanin and bacteria to form a tight mesh. Its hardness and elasticity provide physical protection to the embryo. In addition, SepECPs also have bacteriostatic antimicrobial activity on GRAM- bacteria. By observing the SepECP / Vibrio aestuarianus complex in SEM, we demonstrated the ability of these proteins to agglomerate bacteria and thus inhibit their growth. These original proteins identified from the outer egg case ensure the survival of the species by providing physical and chemical protection to the embryos released in the environment without any maternal protection.

Published
2015
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8. Cellular effects of bacterial N-3-Oxo-dodecanoyl-L-Homoserine lactone on the sponge Suberites domuncula (Olivi, 1792): insights into an intimate inter-kingdom dialogue.

Author

Johan Gardères, Joël Henry, Benoit Bernay, Andrès Ritter, Céline Zatylny-Gaudin, Matthias Wiens, Werner E G Müller, and Gaël Le Pennec

Subjects
Medicine, Science
Abstract

Sponges and bacteria have lived together in complex consortia for 700 million years. As filter feeders, sponges prey on bacteria. Nevertheless, some bacteria are associated with sponges in symbiotic relationships. To enable this association, sponges and bacteria are likely to have developed molecular communication systems. These may include molecules such as N-acyl-L-homoserine lactones, produced by Gram-negative bacteria also within sponges. In this study, we examined the role of N-3-oxododecanoyl-L-homoserine lactone (3-oxo-C12-HSL) on the expression of immune and apoptotic genes of the host sponge Suberites domuncula. This molecule seemed to inhibit the sponge innate immune system through a decrease of the expression of genes coding for proteins sensing the bacterial membrane: a Toll-Like Receptor and a Toll-like Receptor Associated Factor 6 and for an anti-bacterial perforin-like molecule. The expression of the pro-apoptotic caspase-like 3/7 gene decreased as well, whereas the level of mRNA of anti-apoptotic genes Bcl-2 Homolog Proteins did not change. Then, we demonstrated the differential expression of proteins in presence of this 3-oxo-C12-HSL using 3D sponge cell cultures. Proteins involved in the first steps of the endocytosis process were highlighted using the 2D electrophoresis protein separation and the MALDI-TOF/TOF protein characterization: α and β subunits of the lysosomal ATPase, a cognin, cofilins-related proteins and cytoskeleton proteins actin, α tubulin and α actinin. The genetic expression of some of these proteins was subsequently followed. We propose that the 3-oxo-C12-HSL may participate in the tolerance of the sponge apoptotic and immune systems towards the presence of bacteria. Besides, the sponge may sense the 3-oxo-C12-HSL as a molecular evidence of the bacterial presence and/or density in order to regulate the populations of symbiotic bacteria in the sponge. This study is the first report of a bacterial secreted molecule acting on sponge cells and regulating the symbiotic relationship.

Published
2014
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9. A complex set of sex pheromones identified in the cuttlefish Sepia officinalis.

Author

Jérémy Enault, Céline Zatylny-Gaudin, Benoît Bernay, Benjamin Lefranc, Jérôme Leprince, Michèle Baudy-Floc'h, and Joël Henry

Subjects
Medicine, Science
Abstract

The cephalopod mollusk Sepia officinalis can be considered as a relevant model for studying reproduction strategies associated to seasonal migrations. Using transcriptomic and peptidomic approaches, we aim to identify peptide sex pheromones that are thought to induce the aggregation of mature cuttlefish in their egg-laying areas.To facilitate the identification of sex pheromones, 576 5'-expressed sequence tags (ESTs) were sequenced from a single cDNA library generated from accessory sex glands of female cuttlefish. Our analysis yielded 223 unique sequences composed of 186 singletons and 37 contigs. Three major redundant ESTs called SPα, SPα' and SPβ were identified as good candidates for putative sex pheromone transcripts and are part of the 87 unique sequences classified as unknown. The alignment of translated SPα and SPα' revealed a high level of conservation, with 98.4% identity. Translation led to a 248-amino acid precursor containing six peptides with multiple putative disulfide bonds. The alignment of SPα-α' with SPβ revealed a partial structural conservation, with 37.3% identity. Translation of SPβ led to a 252-amino acid precursor containing five peptides. The occurrence of a signal peptide on SPα, SPα' and SPβ showed that the peptides were secreted. RT-PCR and mass spectrometry analyses revealed a co-localization of transcripts and expression products in the oviduct gland. Preliminary in vitro experiments performed on gills and penises revealed target organs involved in mating and ventilation.The analysis of the accessory sex gland transcriptome of Sepia officinalis led to the identification of peptidic sex pheromones. Although preliminary functional tests suggested the involvement of the α3 and β2 peptides in ventilation and mating stimulation, further functional investigations will make it possible to identify the complete set of biological activities expected from waterborne pheromones.

Published
2012
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11. Structural and functional characterization of orcokinin B-like neuropeptides in the cuttlefish ($Sepia\ officinalis$)

Author

Maxime Endress, Céline Zatylny-Gaudin, Jérôme Leprince, Benjamin Lefranc, Erwan Corre, Gildas Le Corguillé, Benoît Bernay, Alexandre Leduc, Jimmy Rangama, Liza Mouret, Anne-Gaëlle Lafont, Arnaud Bondon, Joël Henry, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-High-tech Research Infrastructures for Life Sciences (HeRacLeS), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Différenciation et communication neuronale et neuroendocrine (DC2N), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (ABIMS), Fédération de recherche de Roscoff (FR2424), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Plateforme Proteogen, SFR ICORE 4206, Normandie Université (NU)-Normandie Université (NU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Centre de recherche sur les Ions, les MAtériaux et la Photonique (CIMAP - UMR 6252), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche sur les Matériaux Avancés (IRMA), Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Atomes, Molécules et Agrégats (AMA), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Maxime Endress PhD fellowship was financed by the Normandy Regional Council., ANR-14-CE02-0020,NEMO,Neuropeptides d'organismes marins(2014), Jonchère, Laurent, and Appel à projets générique - Neuropeptides d'organismes marins - - NEMO2014 - ANR-14-CE02-0020 - Appel à projets générique - VALID

Subjects
egg-laying, neuropeptides, Pharmaceutical Science, biological activity, cephalopods, 3D structure, Sepia officinalis, NMR, neurohormones, immunocytochemistry, [CHIM] Chemical Sciences, Drug Discovery, [CHIM]Chemical Sciences, Pharmacology, Toxicology and Pharmaceutics (miscellaneous)
Abstract

International audience; The cuttlefish (Sepia officinalis) is a Cephalopod mollusk that lives in the English Channel and breeds in coastal spawning grounds in spring. A previous work showed that the control of egg-laying is monitored by different types of regulators, among which neuropeptides play a major role. They are involved in the integration of environmental cues, and participate in the transport of oocytes in the genital tract and in the secretion of capsular products. This study addresses a family of neuropeptides recently identified and suspected to be involved in the control of the reproduction processes. Detected by mass spectrometry and immunocytochemistry in the nerve endings of the accessory sex glands of the females and ovary, these neuropeptides are also identified in the hemolymph of egg-laying females demonstrating that they also have a hormone-like role. Released in the hemolymph by the sub-esophageal mass, a region that innervates the genital tract and the neurohemal area of the vena cava, in in vitro conditions these neuropeptides modulated oocyte transport and capsular secretion. Finally, in silico analyses indicated that these neuropeptides, initially called FLGamide, had extensive structural homology with orcokinin B, which motivated their name change.

Published
2022

12. Involvement of GLWamide neuropeptides in polyp contraction of the adult stony coral Euphyllia ancora

Author

Tai-Yu Liu, Ching-Fong Chang, Yan Zhang, Yi-Ling Chiu, Pin-Hsuan Tsai, Shinya Shikina, Céline Zatylny-Gaudin, Yi-ChenYao, National Taiwan Ocean University (NTOU), Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), and Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA)

Subjects
DNA, Complementary, Tentacle, Physiology, Coral, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Science, Glycine, Neuropeptide, Article, Preprohormone, 03 medical and health sciences, 0302 clinical medicine, Leucine, Complementary DNA, medicine, Animals, Amino Acid Sequence, Peptide sequence, 030304 developmental biology, Neurons, 0303 health sciences, Multidisciplinary, biology, Neuropeptides, Tryptophan, Anthozoa, biology.organism_classification, Cell biology, Euphyllia ancora, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, Medicine, medicine.symptom, 030217 neurology & neurosurgery, Neuroscience, Muscle contraction
Abstract

The existence and function of neurons remain largely unexplored in scleractinian corals. To gain a better understanding of neuronal functions in coral physiology, this study focused on Glycine-Leucine-Tryptophan-amide family neuropeptides (GLWamides), which have been shown to induce muscle contraction and larval metamorphosis in other cnidarians. Molecular identification and functional characterization of GLWamides in the adult stony coral Euphyllia ancora were performed. We successfully elucidated the full-length cDNA of GLWamide preprohormone in E. ancora (named EaGLW preprohormone). The deduced amino acid sequence was predicted to contain six potential GLWamide peptides. Tissue distribution analysis demonstrated that transcripts of EaGLW preprohormone were mainly expressed in the mouth (including the pharynx) and tentacles of the polyps. Immunodetection with an anti-GLWamide monoclonal antibody revealed that GLWamide neurons were mainly distributed in the epidermis of the mouth region and tentacle, in agreement with the distribution patterns of the transcripts. Treatment of the isolated mouth and tentacles with synthetic GLWamide peptides induced the contraction of these isolated tissues. Treatment of polyps with synthetic GLWamide peptides induced the contraction of polyps. These results suggest that GLWamides are involved in polyp contraction (myoactivity) in adult scleractinians. Our data provide new information on the physiological function of neuropeptides in scleractinians.

Published
2020

13. Multifaceted roles of the egg perivitelline layer in avian reproduction: Functional insights from the proteomes of chicken egg inner and outer sublayers

Author

Mégane Brégeon, Daniel Tomas, Benoît Bernay, Céline Zatylny-Gaudin, Sonia Georgeault, Valérie Labas, Sophie Réhault-Godbert, Nicolas Guyot, Biologie des Oiseaux et Aviculture (BOA), Université de Tours (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Plate-forme Phénotypage par imagerie in/ex vivo de l'Animal à la Molécule (Plate-forme PIXANIM), Interactions Cellules Organismes Environnement (ICORE), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Plateforme des Microscopies, Université de Tours (UT), This work was supported by the Région Centre-Val de Loire [grant number 2017-119983] and the Agence Nationale de la Recherche [grant number ANR-19-CE21-0006]., and ANR-19-CE21-0006,EQLIPSE,Amélioration de la qualité et des défenses antimicrobiennes internes de l'oeuf(2019)

Subjects
Proteomics, Sperm-Ovum Interactions, Outer perivitelline layer, Proteome, [SDV]Life Sciences [q-bio], Biophysics, Functional annotation, Gallus gallus, Biochemistry, Chicken, stomatognathic diseases, Inner perivitelline layer, Ultrastructure, Tandem Mass Spectrometry, Egg, Animals, Female, sense organs, Gene expression, Avian reproduction, Chickens, Ovum
Abstract

The avian egg perivitelline layer (PL) is a proteinaceous structure that encloses the egg yolk. It consists of the inner and the outer perivitelline layers (IPL and OPL, respectively) that are assumed to play distinct roles in bird reproduction. To gain insight into their respective function, we analyzed the proteome of IPL and OPL in chicken unfertilized eggs after mechanical separation, using a GeLC-MS/MS strategy. Of the 412 proteins identified, 173 proteins were uniquely recovered in IPL and 98 proteins in OPL, while 141 proteins were identified in both sublayers. Genes coding the most abundant proteins were shown to be expressed either in the liver/ovary (IPL formation) or in the oviduct (OPL formation), but rarely in both. The presence of oviduct-specific proteins (including LYZ, VMO1, AvBD11, PTN, OVAL and LOC10175704) in IPL strongly suggests that they participate in the physical association of IPL to OPL, whose tight attachment was further evidenced by analyses of IPL/OPL interfaces (by scanning electron microscopy). Functional annotation of identified proteins revealed functions associated with fertilization and early development for IPL, while OPL would rather participate in egg defense and embryogenesis. Collectively, our data highlight the complementary functions of IPL and OPL that are major determinants of bird reproductive success. SIGNIFICANCE: The present study unveils for the first time the individual proteomes of the two sublayers composing the chicken egg perivitelline layer (PL), which allowed to assign their respective putative biological roles in avian reproduction. The combination of proteomics with gene expression and ultrastructural analyses provides insightful data on the structure and biochemistry of the avian PL. The functional annotation of PL proteins highlights the multifaceted biological functions of this structure in reproduction including fertilization, embryonic development, and antimicrobial protection. This work will stimulate further research to validate predicted functions and to compare the physiology and the functional specificities of PL in egg-laying species.

Published
2021

14. Involvement of RFamide neuropeptides in polyp contraction of the adult scleractinian corals Euphyllia ancora and Stylophora pistillata

Author

Yu-Ying Ho, Sylvie Dufour, Céline Zatylny-Gaudin, Yi-Ling Chiu, Ching-Fong Chang, Jack I-Chen Yao, Yan Zhang, and Shinya Shikina

Subjects
biology, Epidermis (botany), Coral, Neuropeptides, Neuropeptide, Stylophora pistillata, biology.organism_classification, Anthozoa, Preprohormone, Euphyllia ancora, Cell biology, Endocrinology, Animals, Animal Science and Zoology, Amino Acid Sequence, FMRFamide, Peptide sequence
Abstract

The distribution and functions of neurons in scleractinian corals remain largely unknown. This study focused on the Arg-Phe amide family of neuropeptides (RFamides), which have been shown to be involved in a variety of biological processes in animals, and performed molecular identification and characterization in the adult scleractinian coral Euphyllia ancora. The deduced amino acid sequence of the identified RFamide preprohormone was predicted to contain 20 potential neuropeptides, including 1 Pro-Gly-Arg-Phe (PGRF) amide and 15 Gln-Gly-Arg-Phe (QGRF) amide peptides. Tissue distribution analysis showed that the level of transcripts in the tentacles was significantly higher than that in other polyp tissues. Immunohistochemical analysis with the FMRFamide antibody showed that RFamide neurons were mainly distributed in the epidermis of the tentacles and mouth with pharynx. Treatment of E. ancora polyps with synthetic QGRFamide peptides induced polyp contraction. The induction of polyp contraction by QGRFamide peptide treatment was also observed in another scleractinian coral, Stylophora pistillata. These results strongly suggested that RFamides play a role in the regulation of polyp contraction in adult scleractinians.

Published
2021

15. Omic Analysis of the

Author

Louis, Benoist, Erwan, Corre, Benoit, Bernay, Joel, Henry, and Céline, Zatylny-Gaudin

Subjects
Proteomics, Sepia, Proteome, Decapodiformes, Animals, Hematopoiesis
Abstract

Cephalopods, like other protostomes, lack an adaptive immune system and only rely on an innate immune system. The main immune cells are haemocytes (Hcts), which are able to respond to pathogens and external attacks. First reports based on morphological observations revealed that the white body (WB) located in the optic sinuses of cuttlefish was the origin of Hcts. Combining transcriptomic and proteomic analyses, we identified several factors known to be involved in haematopoiesis in vertebrate species in cuttlefish WB. Among these factors, members of the JAK-STAT signaling pathway were identified, some of them for the first time in a molluscan transcriptome and proteome. Immune factors, such as members of the Toll/NF-κB signaling pathway, pattern recognition proteins and receptors, and members of the oxidative stress responses, were also identified, and support an immune role of the WB. Both transcriptome and proteome analyses revealed that the WB harbors an intense metabolism concurrent with the haematopoietic function. Finally, a comparative analysis of the WB and Hct proteomes revealed many proteins in common, confirming previous morphological studies on the origin of Hcts in cuttlefish. This molecular work demonstrates that the WB is multifunctional and provides bases for haematopoiesis regulation in cuttlefish.

Published
2020

16. Identification and structural characterization of the factors involved in vitellogenesis and its regulation in the African Osteoglossiforme of aquacultural interest Heterotis niloticus (Cuvier, 1829)

Author

Joël Henry, Benoit Bernay, Julien Pontin, Céline Zatylny-Gaudin, Julie Orjuela, N'Zi Daniel Koua, Marie-Pierre Dubos, Erwan Corre, Jésus Núñez-Rodriguez, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Institut de Recherche pour le Développement (IRD), Normandie Université (NU), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (ABIMS), Fédération de recherche de Roscoff (FR2424), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Station biologique de Roscoff (SBR), and Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
endocrine system, Pituitary gland, Proteome, [SDV]Life Sciences [q-bio], Estrogen receptor, 030209 endocrinology & metabolism, Ovary, Receptors, Cell Surface, Vitellogenin, Aquaculture, Protein Structure, Secondary, Neurohormones, Andrology, 03 medical and health sciences, Vitellogenins, 0302 clinical medicine, Endocrinology, medicine, Animals, Computer Simulation, 14. Life underwater, Amino Acid Sequence, Heterotis niloticus, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, biology, Base Sequence, Reproduction, [SDV.BA]Life Sciences [q-bio]/Animal biology, Vitellogenesis, Fishes, Estradiol receptors, Prolactin, Mucus, medicine.anatomical_structure, Liver, biology.protein, Animal Science and Zoology, Female, Transcriptome, Luteinizing hormone
Abstract

The African bonytongue (Heterotis niloticus) is an excellent candidate for fish farming because it has outstanding biological characteristics and zootechnical performances. However, the absence of sexual dimorphism does not favor its reproduction in captivity or the understanding of its reproductive behavior. Moreover, no molecular data related to its reproduction is yet available. This study therefore focuses on the structural identification of the different molecular actors of vitellogenesis expressed in the pituitary gland, the liver and the ovary of H. niloticus. A transcriptomic approach based on de novo RNA sequencing of the pituitary gland, ovary and liver of females in vitellogenesis led to the creation of three transcriptomes. In silico analysis of these transcriptomes identified the sequences of pituitary hormones such as prolactin (PRL), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) and their ovarian receptors (PRLR, FSHR, LHR). In the liver and ovary, estrogen receptors (ER) beta and gamma, liver vitellogenins (VtgB and VtgC) and their ovarian receptors (VLDLR) were identified. Finally, the partial transcript of an ovarian Vtg weakly expressed compared to hepatic Vtg was identified based on structural criteria. Moreover, a proteomic approach carried out from mucus revealed the presence of one Vtg exclusively in females in vitellogenesis. In this teleost fish that does not exhibit sexual dimorphism, mucus Vtg could be used as a sexing biomarker based on a non-invasive technique compatible with the implementation of experimental protocols in vivo.

Published
2020

17. Omic Analysis of the Sepia officinalis White Body: New Insights into Multifunctionality and Haematopoiesis Regulation

Author

Benoit Bernay, Céline Zatylny-Gaudin, Louis Benoist, Erwan Corre, Joël Henry, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (ABIMS), Fédération de recherche de Roscoff (FR2424), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Station biologique de Roscoff (SBR), and Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Cuttlefish, Innate immune system, 030102 biochemistry & molecular biology, white body, sepia officinalis, General Chemistry, Biology, haematopoiesis, Acquired immune system, Biochemistry, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Cell biology, JAK-STAT pathway, Transcriptome, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, 03 medical and health sciences, Haematopoiesis, 030104 developmental biology, Immune system, mollusc, Proteome, Sepia
Abstract

International audience; Cephalopods, like other protostomes, lack an adaptive immune system and only rely on an innate immune system. The main immune cells are haemocytes (Hcts), which are able to respond to pathogens and external attacks. First reports based on morphological observations revealed that the white body (WB) located in the optic sinuses of cuttlefish was the origin of Hcts. Combining transcriptomic and proteomic analyses, we identified several factors known to be involved in haematopoiesis in vertebrate species in cuttlefish WB. Among these factors, members of the JAK-STAT signaling pathway were identified, some of them for the first time in a molluscan transcriptome and proteome. Immune factors, such as members of the Toll/NF-κB signaling pathway, pattern recognition proteins and receptors, and members of the oxidative stress responses, were also identified, and support an immune role of the WB. Both transcriptome and proteome analyses revealed that the WB harbors an intense metabolism concurrent with the haematopoietic function. Finally, a comparative analysis of the WB and Hct proteomes revealed many proteins in common, confirming previous morphological studies on the origin of Hcts in cuttlefish. This molecular work demonstrates that the WB is multifunctional and provides bases for haematopoiesis regulation in cuttlefish

Published
2020

18. Crustacean cardioactive peptides: Expression, localization, structure, and a possible involvement in regulation of egg-laying in the cuttlefish Sepia officinalis

Author

Maxime Endress, Jimmy Rangama, Jérôme Leprince, Benoit Bernay, Joël Henry, Anne-Gaëlle Lafont, Benjamin Lefranc, Arnaud Bondon, Gildas Le Corguillé, Louis Benoist, Alexandre Leduc, Erwan Corre, Céline Zatylny-Gaudin, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (ABIMS), Fédération de recherche de Roscoff (FR2424), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Centre de recherche sur les Ions, les MAtériaux et la Photonique (CIMAP - UMR 6252), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche sur les Matériaux Avancés (IRMA), Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU), Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Caen Normandie (UNICAEN), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA), and Jonchère, Laurent

Subjects
Central Nervous System, Male, 0301 basic medicine, Cuttlefish, Sepia, animal structures, Oviposition, Neuropeptide, Zoology, Context (language use), Oviducts, Sepia officinalis, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, 03 medical and health sciences, Endocrinology, Animals, Tissue Distribution, Amino Acid Sequence, 14. Life underwater, Peptide sequence, [SDV.BDLR.RS] Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, biology, Reproduction, egg-laying, Neuropeptides, CCAPs, fungi, Decapodiformes, Gene Expression Regulation, Developmental, biology.organism_classification, NMR, Cephalopod, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, 030104 developmental biology, Oviparity, [SDV.BA.ZI] Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, Oocytes, Oviduct, Female, Animal Science and Zoology, Sequence Alignment, Free nerve ending
Abstract

International audience; The cuttlefish (Sepia officinalis) is a cephalopod mollusk distributed on the western European coast, in the West African Ocean and in the Mediterranean Sea. On the Normandy coast (France), cuttlefish is a target species of professional fishermen, so its reproduction strategy is of particular interest in the context of stock management. Egg-laying, which is coastal, is controlled by several types of regulators among which neuropeptides. The cuttlefish neuropeptidome was recently identified by Zatylny-Gaudin et al. (2016). Among the 38 neuropeptide families identified, some were significantly overexpressed in egg-laying females as compared to mature males. This study is focused on crustacean cardioactive peptides (CCAPs), a highly expressed neuropeptide family strongly suspected of being involved in the control of egg-laying. We investigated the functional and structural characterization and tissue mapping of CCAPs, as well as the expression patterns of their receptors. CCAPs appeared to be involved in oocyte transport through the oviduct and in mechanical secretion of capsular products. Immunocytochemistry revealed that the neuropeptides were localized throughout the central nervous system (CNS) and in the nerve endings of the glands involved in egg-capsule synthesis and secretion, i.e. the oviduct gland and the main nidamental glands. The CCAP receptor was expressed in these glands and in the subesophageal mass of the CNS. Multiple sequence alignments revealed a high level of conservation of CCAP protein precursors in Sepia officinalis and Loligo pealei, two cephalopod decapods. Primary sequences of CCAPs from the two species were fully conserved, and cryptic peptides detected in the nerve endings were also partially conserved, suggesting biological activity that remains unknown for the time being.

Published
2018

20. Behavior of Antimicrobial Peptide K4 in a Marine Environment

Author

L Benoist, Valérie Cornet, Céline Zatylny-Gaudin, E Vetois, Julien Pontin, Benoit Bernay, Baptiste Houyvet, Joël Henry, Alexandre Leduc, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), SATMAR, Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, and Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)

Subjects
0301 basic medicine, Aquatic Organisms, Oyster, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV]Life Sciences [q-bio], 030106 microbiology, Aquaculture, Microbiology, 03 medical and health sciences, Antibiotics, biology.animal, 14. Life underwater, Molecular Biology, Vibrio, biology, Chemistry, [SDV.BA]Life Sciences [q-bio]/Animal biology, biology.organism_classification, Antimicrobial, Hatchery, Biodegradability, Aeromonas salmonicida, 030104 developmental biology, Molecular Medicine, Dicentrarchus, Aeromonas, Artemia salina, Water Microbiology, Antimicrobial peptide, Bacteria, Antimicrobial Cationic Peptides
Abstract

International audience; K4 is a de novo peptide with antibacterial activity on human pathogens. It has a short sequence (14 amino acids), with a cationic N-terminal moiety and an amphipathic ɑ-helix structure. The present paper demonstrates its activity on Vibrio bacteria in a marine environment. It was found non-toxic on marine organisms including Artemia salina, Dicentrarchus labrax, and Magallana gigas at different developmental stages, but influenced the growth of unicellular organisms like microalgae, depending on the algal strain and on K4 concentration. Furthermore, an original approach coupling liquid chromatography (RP-HPLC) and mass spectrometry (MS/MS) allowed us to monitor the degradation time course of the peptide for the first time in conditions close to a hatchery environment, i.e., in the presence of oyster spat. We detected truncated forms over time, and the full K4 was gradually no longer found in these filter-feeder oysters. Finally, using an automated optical density meter, we monitored the growth of several aquatic bacteria identified as pathogenic on animals. K4 had a bactericidal effect on Aeromonas salmonicida and Vibrio splendidus LGP32 at concentrations below 45 μg mL-1. Our results show that K4 could be an environment-friendly alternative to antibiotics, non-toxic to several marine organisms. The use of K4 would be particularly useful to decrease the bacterial load associated with food intake in the early developmental stages of marine animals reared in hatcheries.

Published
2019

21. Design of antimicrobial peptides from a cuttlefish database

Author

Gildas Le Corguillé, Erwan Corre, Baptiste Houyvet, Céline Zatylny-Gaudin, Bruno Zanuttini, Joël Henry, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), SATMAR, Equipe MAD - Laboratoire GREYC - UMR6072, Groupe de Recherche en Informatique, Image et Instrumentation de Caen (GREYC), Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), Station biologique de Roscoff (SBR), and Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Signal peptide, Design, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV]Life Sciences [q-bio], Clinical Biochemistry, Antimicrobial peptides, Peptide, Predictive tools, computer.software_genre, Sepia officinalis, Biochemistry, 03 medical and health sciences, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Sepia, Databases, Protein, Vibrio alginolyticus, chemistry.chemical_classification, biology, Database, Bacteria, Organic Chemistry, Decapodiformes, biology.organism_classification, Antimicrobial, 3. Good health, Aeromonas salmonicida, 030104 developmental biology, chemistry, Antibacterial activity, Transcriptome, computer, Antimicrobial Cationic Peptides
Abstract

International audience; No antimicrobial peptide has been identified in cephalopods to date. Annotation of transcriptomes or genomes using basic local alignment Search Tool failed to yield any from sequence identities. Therefore, we searched for antimicrobial sequences in the cuttlefish (Sepia officinalis) database by in silico analysis of a transcriptomic database. Using an original approach based on the analysis of cysteine-free antimicrobial peptides selected from our Antimicrobial Peptide Database (APD3), the online prediction tool of the Collection of Anti-Microbial Peptides (CAMPR3), and a homemade software program, we identified potential antibacterial sequences. Nine peptides less than 25 amino acids long were synthesized. The hydrophobic content of all nine of them ranged from 30 to 70%, and they could form alpha-helices. Three peptides possessed similarities with piscidins, one with BMAP-27, and five were totally new. Their antibacterial activity was evaluated on eight bacteria including the aquatic pathogens Vibrio alginolyticus, Aeromonas salmonicida, or human pathogens such as Salmonella typhimurium, Listeria monocytogenes, or Staphylococcus aureus. Despite the prediction of an antimicrobial potential for eight of the peptides, only two-GR21 and KT19-inhibited more than one bacterial strain with minimal inhibitory concentrations below 25 µM. Some sequences like VA20 and FK19 were hemolytic, while GR21 induced less than 10% of hemolysis on human blood cells at a concentration of 200 µM. GR21 was the only peptide derived from a precursor with a signal peptide, suggesting a real role in cuttlefish immune defense.

Published
2018

22. Egg-Laying in the Cuttlefish Sepia officinalis

Author

Céline Zatylny-Gaudin, Joël Henry, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Centre de recherche en gestion (CRG), and Centre National de la Recherche Scientifique (CNRS)-École polytechnique (X)

Subjects
Cuttlefish, 0303 health sciences, Zoology, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Biology, Egg laying, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, 03 medical and health sciences, 0302 clinical medicine, Officinalis, Sepia, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, GeneralLiterature_REFERENCE(e.g.,dictionaries,encyclopedias,glossaries), 030217 neurology & neurosurgery, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology
Abstract

International audience

Published
2018

23. Identification of a moronecidin-like antimicrobial peptide in the venomous fish Pterois volitans: Functional and structural study of pteroicidin-α

Author

Claude Bouchon, Céline Zatylny-Gaudin, Yolande Bouchon-Navaro, Benoit Bernay, Baptiste Houyvet, Erwan Corre, Didier Goux, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), SATMAR, Centre de Microscopie Appliquée à la Biologie [Caen] (CMABio3), Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), Station biologique de Roscoff (SBR), and Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Fish Proteins, 0301 basic medicine, Staphylococcus aureus, Pterois, [SDV]Life Sciences [q-bio], Antimicrobial peptides, Peptide, Aeromonas salmonicida, Aquatic Science, Microbiology, 03 medical and health sciences, Escherichia coli, medicine, Animals, Environmental Chemistry, Sea bass, chemistry.chemical_classification, 030102 biochemistry & molecular biology, biology, Fishes, General Medicine, Venomous fish, biology.organism_classification, medicine.disease, Antimicrobial, Hemolysis, Anti-Bacterial Agents, 030104 developmental biology, chemistry, [SDV.IMM]Life Sciences [q-bio]/Immunology, Antimicrobial Cationic Peptides
Abstract

International audience; The present study characterizes for the first time an antimicrobial peptide in lionfish (Pterois volitans), a venomous fish. Using a peptidomic approach, we identified a mature piscidin in lionfish and called it pteroicidin-α. We detected an amidated form (pteroicidin-α- CONH2) and a non-amidated form (pteroicidin-α-COOH), and then performed their functional and structural study. Interestingly, the two peptides displayed different antibacterial and hemolytic activity levels. Pteroicidin-α-CONH2 was bactericidal on human pathogens like Staphylococcus aureus or Escherichia coli, as well as on the fish pathogen Aeromonas salmonicida, while pteroicidin-α-COOH only inhibited their growth. Furthermore, the two peptides induced hemolysis of red blood cells from different vertebrates, namely humans, sea bass and lesser-spotted dogfish. Hemolysis occurred with low concentrations of pteroicidin-α-CONH2, indicating greater toxicity of the amidated form. Circular dichroism analysis showed that both peptides adopted a helical conformation, yet with a greater α-helix content in pteroicidin-α-CONH2. Overall, these results suggest that amidation strongly influences pteroicidin-α by modifying its structure and its physico-chemical characteristics and by increasing its hemolytic activity.

Published
2018

24. Protecting honey bees: identification of a new varroacide by in silico, in vitro, and in vivo studies

Author

Romain Bonafos, Céline Zatylny-Gaudin, Fabienne Dulin, Ronan Bureau, Céline Ballandonne, Bertrand Guillet, Marie Pierre Halm, Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Normandie Université (NU), Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Musée d'Histoire de Nantes -- Château des ducs de Bretagne, and Mairie de Nantes

Subjects
Models, Molecular, Varroidae, In silico, Molecular Sequence Data, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Pirimicarb, chemistry.chemical_compound, In vivo, Animals, Destructor, Amino Acid Sequence, Acaricides, ComputingMilieux_MISCELLANEOUS, Genetics, Base Sequence, General Veterinary, biology, business.industry, Acaricide, General Medicine, Bees, biology.organism_classification, Biotechnology, Molecular Docking Simulation, Pyrimidines, Infectious Diseases, chemistry, Docking (molecular), Insect Science, Varroa destructor, [SDE]Environmental Sciences, Acetylcholinesterase, Parasitology, Varroa, Carbamates, Cholinesterase Inhibitors, business, Sequence Alignment
Abstract

Varroa destructor is the main concern related to the gradual decline of honeybees. Nowadays, among the various acaricides used in the control of V. destructor, most presents increasing resistance. An interesting alternative could be the identification of existent molecules as new acaricides with no effect on honeybee health. We have previously constructed the first 3D model of AChE for honeybee. By analyzing data concerning amino acid mutations implicated in the resistance associated to pesticides, it appears that pirimicarb should be a good candidate for varroacide. To check this hypothesis, we characterized the AChE gene of V. destructor. In the same way, we proposed a 3D model for the AChE of V. destructor. Starting from the definition of these two 3D models of AChE in honeybee and varroa, a comparison between the gorges of the active site highlighted some major differences and particularly different shapes. Following this result, docking studies have shown that pirimicarb adopts two distinct positions with the strongest intermolecular interactions with VdAChE. This result was confirmed with in vitro and in vivo data for which a clear inhibition of VdAChE by pirimicarb at 10 μM (contrary to HbAChE) and a 100% mortality of varroa (dose corresponding to the LD50 (contact) for honeybee divided by a factor 100) were observed. These results demonstrate that primicarb could be a new varroacide candidate and reinforce the high relationships between in silico, in vitro, and in vivo data for the design of new selective pesticides.

Published
2014

25. Dual role of the cuttlefish salivary proteome in defense and predation

Author

Bruno Zanuttini, Erwan Corre, Valérie Cornet, Céline Zatylny-Gaudin, Joël Henry, Gildas Le Corguillé, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Equipe MAD - Laboratoire GREYC - UMR6072, Groupe de Recherche en Informatique, Image et Instrumentation de Caen (GREYC), Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), and Normandie Université (NU)

Subjects
Proteomics, Cuttlefish, Saliva, Proteome, [SDV]Life Sciences [q-bio], In silico, Biophysics, Venom, Biology, Biochemistry, Transcriptome, 03 medical and health sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Salivary Proteins and Peptides, 030304 developmental biology, 0303 health sciences, Ecology, [SDV.BA]Life Sciences [q-bio]/Animal biology, 030302 biochemistry & molecular biology, Decapodiformes, RNA, Predatory Behavior, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Electrophoresis, Polyacrylamide Gel
Abstract

International audience; We characterized the proteome of the posterior salivary glands of the cephalopod S. officinalis by combining de novo RNA sequencing and mass spectrometry. In silico analysis of the transcriptome revealed the occurrence of three main categories of proteins: enzymes, immune factors and toxins. Protein identification by SDS-PAGE and MALDI-TOF/TOF confirmed the occurrence of proteins essential to venom-like enzymes: peptidase S1 under four isoforms, phospholipase A2 and two toxins. The first toxin is a cystein rich secreted protein (CRISP), a common toxin found in all venomous animals. The second one is cephalotoxin, which is specific to decabrachia cephalopods. Secretions of the posterior salivary glands are transported to the cephalopodium; they are involved in prey catching but also in gamete storage, fertilization and egg-laying. The paralyzing activity and the antimicrobial effect of saliva suggest a dual role in predation and in immune defense in cuttlefish.BIOLOGICAL SIGNIFICANCE:The originality of this study lies in the use of a transcriptomic approach (de novo RNA sequencing) coupled to a proteomic approach to get an overview of posterior salivary glands in S. officinalis. In cephalopods, these glands are involved in predation, more precisely in paralyzing preys and digesting them. Our in silico analysis equally reveals a role in immune defense as observed in mammals' saliva. Our study also shows the specificity of cuttlefish venom, with the identification of cephalotoxins, proteins that are not found in octopuses. Finally, we show that cuttlefish saliva is a complex mixture that has antibacterial and crippling properties, but no lethal effect.

Published
2014

26. Neuropeptidome of the Cephalopod Sepia officinalis : Identification, Tissue Mapping, and Expression Pattern of Neuropeptides and Neurohormones during Egg Laying

Author

Joël Henry, Céline Zatylny-Gaudin, Benoit Bernay, Bruno Zanuttini, Alexandra Kraut, Erwan Corre, Alexandre Leduc, Yohan Couté, Johan Gardères, Gildas Le Corguillé, Valérie Cornet, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Biologie des mollusques marins et des écosystèmes associés (BioMEA), Normandie Université (NU)-Normandie Université (NU)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS), SAPHYN (SAnté et PHYsique Nucléaire), Equipe MAD - Laboratoire GREYC - UMR6072, Groupe de Recherche en Informatique, Image, Automatique et Instrumentation de Caen (GREYC), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Station biologique de Roscoff [Roscoff] (SBR), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Interactions Cellules Organismes Environnement (ICORE), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU), Laboratoire de Biotechnologie et Chimie Marines (LBCM), Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS)-Université de Bretagne Sud (UBS), Etude de la dynamique des protéomes (EDyP), Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Centre de recherche en gestion (CRG), École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Groupe de Recherche en Informatique, Image et Instrumentation de Caen (GREYC), Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Université de Bretagne Sud (UBS)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS), Etude de la dynamique des protéomes (EDyP ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Sorbonne Université (SU)-Université des Antilles (UA)-Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut de Recherche pour le Développement (IRD), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), Laboratoire d'étude de la dynamique des protéomes (LEDyP), Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (ABIMS), Fédération de recherche de Roscoff (FR2424), and Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Station biologique de Roscoff (SBR)

Subjects
0301 basic medicine, medicine.medical_specialty, Sepia, Proteome, Oviposition, In silico, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Neuropeptide, Biology, Biochemistry, Transcriptome, 03 medical and health sciences, Internal medicine, Hemolymph, medicine, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, RNA, Messenger, cuttlefish, brain, egg-laying neuropeptidome, transcriptome, neurohormones, Illumina dye sequencing, ComputingMilieux_MISCELLANEOUS, Neurotransmitter Agents, Neuropeptides, Molecular Sequence Annotation, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, General Chemistry, Cell biology, 030104 developmental biology, Endocrinology, Organ Specificity, Female, Neurohormones
Abstract

International audience; Cephalopods exhibit a wide variety of behaviors such as prey capture, communication, camouflage, and reproduction thanks to a complex central nervous system (CNS) divided into several functional lobes that express a wide range of neuropeptides involved in the modulation of behaviors and physiological mechanisms associated with the main stages of their life cycle. This work focuses on the neuropeptidome expressed during egg-laying through de novo construction of the CNS transcriptome using an RNAseq approach (Illumina sequencing). Then, we completed the in silico analysis of the transcriptome by characterizing and tissue-mapping neuropeptides by mass spectrometry. To identify neuropeptides involved in the egg-laying process, we determined (1) the neuropeptide contents of the neurohemal area, hemolymph (blood), and nerve endings in mature females and (2) the expression levels of these peptides. Among the 38 neuropeptide families identified from 55 transcripts, 30 were described for the first time in Sepia officinalis, 5 were described for the first time in the animal kingdom, and 14 were strongly overexpressed in egg-laying females as compared with mature males. Mass spectrometry screening of hemolymph and nerve ending contents allowed us to clarify the status of many neuropeptides, that is, to determine whether they were neuromodulators or neurohormones.

Published
2015

27. Evidence of Antibacterial Activities in Peptide Fractions Originating from Snow Crab (Chionoecetes opilio) By-Products

Author

Richard Saint-Louis, Céline Zatylny-Gaudin, Lucie Beaulieu, Sharon ThibaultS. Thibault, Jacinthe Thibodeau, and Michel Desbiens

Subjects
chemistry.chemical_classification, Aeromonas caviae, biology, Vibrio parahaemolyticus, Peptide, Vibrio vulnificus, biology.organism_classification, Microbiology, Listonella anguillarum, Amino acid, chemistry, Biochemistry, Molecular Medicine, Morganella morganii, Molecular Biology, Bacteria
Abstract

Antibacterial peptide fractions generated via proteolytic processing of snow crab by-products exhibited activity against Gram-negative and Gram-positive bacteria. Among the bacterial strains tested, peptide fractions demonstrated inhibitory activity against the Gram-negative bacteria such as Aeromonas caviae, Aeromonas hydrophila, Campylobacter jejuni, Listonella anguillarum, Morganella morganii, Shewanella putrefasciens, Vibrio parahaemolyticus and Vibrio vulnificus and against a few Gram-positive bacteria such as Listeria monocytogenes, Staphylococcus epidermidis and Streptococcus agalactiae. The principal bioactive peptide fraction was comprised mainly of proteins and minerals (74.3 and 15.5%, respectively). Lipids were not detected. The amino acid content revealed that arginine (4.6%), glutamic acid (5.3%) and tyrosine (4.8%) residues were represented in the highest composition in the antibacterial peptide fraction. The optimal inhibitory activity was observed at alkaline pH. The V. vulnificus strain, most sensitive to the peptide fraction, was used to develop purification methods. The most promising chromatography resins selected for purification, in order to isolate peptides of interest and to carry out their detailed biochemical characterization, were the SP-Sepharose™ Fast Flow cation exchanger and the Phenyl Sepharose™ High Performance hydrophobic interaction media. The partially purified antibacterial peptide fraction was analyzed for minimum inhibitory concentration (MIC) determination, and the value obtained was 25 μg ml−1. Following mass spectrometry analysis, the active peptide fraction seems to be a complex of molecules comprised of several amino acids and other organic compounds. In addition, copper was the main metal found in the active peptide fraction. Results indicate the production of antibacterial molecules from crustacean by-products that support further applications for high-value bioproducts in several areas such as food and health.

Published
2010

28. Characterization of a novel LFRFamide neuropeptide in the cephalopod Sepia officinalis

Author

Jérôme Leprince, Hubert Vaudry, Céline Zatylny-Gaudin, Joël Henry, Benoit Bernay, Bruno Zanuttini, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Laboratoire de Physiogénomique / NeuroTranscriptomes et Paléogénétique (LPG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Equipe MAD - Laboratoire GREYC - UMR6072, Groupe de Recherche en Informatique, Image et Instrumentation de Caen (GREYC), Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU), Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Différenciation et communication neuronale et neuroendocrine (DC2N), Centre de recherche en gestion (CRG), Centre National de la Recherche Scientifique (CNRS)-École polytechnique (X), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), and École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Central Nervous System, Male, Cuttlefish, Sepia, Physiology, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Amino Acid Sequence, MESH: Muscle, Smooth, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Oviducts, MESH: Neuropeptides, Biochemistry, 0302 clinical medicine, Endocrinology, MESH: Rectum, MESH: FMRFamide, Tandem Mass Spectrometry, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Hemolymph, MESH: Animals, Peptide sequence, Nerve Endings, 0303 health sciences, MESH: Sepia, MESH: Nerve Endings, Muscle Tonus, MESH: Oligopeptides, Officinalis, Female, MESH: Animal Structures, Oligopeptides, Spectrometry, Mass, Electrospray Ionization, medicine.medical_specialty, Neuropeptide, Biology, MESH: Spectrometry, Mass, Electrospray Ionization, 03 medical and health sciences, Cellular and Molecular Neuroscience, Internal medicine, medicine, MESH: Central Nervous System, Animals, Amino Acid Sequence, FMRFamide, 030304 developmental biology, Neuropeptides, Rectum, MESH: Tandem Mass Spectrometry, Animal Structures, Muscle, Smooth, MESH: Muscle Tonus, biology.organism_classification, MESH: Male, Cephalopod, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, MESH: Female, MESH: Oviducts, 030217 neurology & neurosurgery
Abstract

International audience; From a single LC-MS/MS analysis, a new C-terminally extended RFamide neuropeptide was characterized in Sepia officinalis. The experimental strategy was based on the specific neutral loss associated with RFamide breakdown. Mass losses of 17 Da (C-terminally amide) and 320 Da (RFamide) have been observed for three known peaks of m/z 581.7 (FLRFamide), 599.8 (FMRFamide), 1096.3 (ALSGDAFLRFamide) and one unknown of m/z 752.8. The primary sequence of the peptide of m/z 752.8 was GNLFRFamide. MS/MS analyses revealed that this novel neuropeptide, called sepFRF1, is largely distributed in the central nervous system of cuttlefish of both sexes. Probably transported in the visceral nerve from the subesophageal mass (the peptide was not detected in the hemolymph), this neuropeptide targeted the rectum in agreement with its peripheral distribution. From concentrations as low as 10(-9)M, sepFRF1 increased the frequency, tonus and amplitude of rectal contractions. SepFRF1 is the first RFamide peptide identified in Sepia officinalis that is not derived from the FaRPs precursor. SepFRF1 could belong to a RFamide subfamily identified in gastropods and may be involved in feeding behavior.

Published
2010

29. The Toll/NF-κB pathway in cuttlefish symbiotic accessory nidamental gland

Author

Gildas Le Corguillé, Joël Henry, Céline Zatylny-Gaudin, Erwan Corre, Valérie Cornet, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), Station biologique de Roscoff (SBR), and Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Sepia, [SDV]Life Sciences [q-bio], Immunology, Nitric Oxide Synthase Type II, Biology, Transcriptome, chemistry.chemical_compound, Immune system, Animals, Genitalia, Symbiosis, Toll-like receptor, Innate immune system, Ecology, Microbiota, Toll-Like Receptors, NF-kappa B, NF-κB, Immunity, Innate, Cell biology, chemistry, [SDV.IMM]Life Sciences [q-bio]/Immunology, Signal transduction, Signal Transduction, Developmental Biology, Nidamental gland
Abstract

International audience; The female genital apparatus of decapod cephalopods contains a symbiotic accessory nidamental gland (ANG) that harbors bacterial symbionts. Although the ANG bacterial consortium is now well described, the impact of symbiosis on Sepia officinalis innate immunity pathways remains unknown. In silico analysis of the de novo transcriptome of ANG highlighted for the first time the existence of the NF-κB pathway in S. officinalis. Several signaling components were identified, i.e. five Toll-like receptors, eight signaling cascade features, and the immune response target gene iNOS, previously described as being involved in the initiation of bacterial symbiosis in a cephalopod gland. This work provides a first key for studying bacterial symbiosis and its impact on innate immunity in S. officinalis ANG.

Published
2015

30. Molecular characterization of peptide fractions of a Tilapia (Oreochromis niloticus) by-product hydrolysate and in vitro evaluation of antibacterial activity

Author

Gildas Le Corguillé, Benoit Bernay, Vincent Fournier, Marie Robert, Erwan Corre, Joël Henry, Céline Zatylny-Gaudin, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), AQUATIV, ABiMS - Informatique et bioinformatique = Analysis and Bioinformatics for Marine Science (FR2424), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, and Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)

Subjects
chemistry.chemical_classification, food.ingredient, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, biology, [SDV]Life Sciences [q-bio], Edwardsiella tarda, Bioengineering, Tilapia, Peptide, biology.organism_classification, Applied Microbiology and Biotechnology, Biochemistry, Hydrolysate, Amino acid, Oreochromis, food, chemistry, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Yersinia ruckeri, human activities, Bacillus megaterium
Abstract

International audience; We investigated the molecular characterization of a Tilapia by-product hydrolysate to asses its nutritional potential in aquafeed. The proximate composition and amino acid composition showed that the hydrolysate had high protein content and contained all the essential amino acids required for carnivorous fish nutrition. The molecular weight profile determined by High Performance Size Exclusion Chromatography (HPSEC) indicated that the hydrolysate was mainly composed of polypeptides (46.9%) and oligopeptides (35.8%). We also characterized the peptide fraction by nLC-ESI-MS/MS and OFFGEL-nLC-MALDI-TOF/TOF. We identified 1374 peptides, and highlighted a high diversity of oligopeptides and polypeptides. Finally, the Tilapia by-product hydrolysate exhibited antimicrobial activity against Yersinia ruckeri and to a lesser extent against Edwardsiella tarda and Bacillus megaterium. These preliminary results suggest the strong nutritional and functional potential of Tilapia by-product hydrolysates.

Published
2015

31. Diversity of the RFamide peptide family in mollusks

Author

Céline Zatylny-Gaudin, Pascal Favrel, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), and Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA)

Subjects
animal structures, [SDV]Life Sciences [q-bio], Endocrinology, Diabetes and Metabolism, Macrocallista nimbosa, Peptide, Genomics, Computational biology, Review Article, Bioinformatics, lcsh:Diseases of the endocrine glands. Clinical endocrinology, Endocrinology, FMRFamide, Receptor, Gene, chemistry.chemical_classification, Mollusks, lcsh:RC648-665, biology, FaRPs, biology.organism_classification, Luqin, chemistry, LFRFamide, Signal transduction, CCK/SK, Neurohormones, NPF
Abstract

International audience; Since the initial characterization of the cardioexcitatory peptide FMRFamide in the bivalve mollusk Macrocallista nimbosa, a great number of FMRFamide-like peptides (FLPs) have been identified in mollusks. FLPs were initially isolated and molecularly characterized in model mollusks using biochemical methods. The development of recombinant technologies and, more recently, of genomics has boosted knowledge on their diversity in various mollusk classes. Today, mollusk FLPs represent approximately 75 distinct RFamide pep-tides that appear to result from the expression of only five genes: the FMRFamide-related peptide gene, the LFRFamide gene, the luqin gene, the neuropeptide F gene, and the chole-cystokinin/sulfakinin gene. FLPs display a complex spatiotemporal pattern of expression in the central and peripheral nervous system. Working as neurotransmitters, neuromodu-lators, or neurohormones, FLPs are involved in the control of a great variety of biological and physiological processes including cardiovascular regulation, osmoregulation, reproduction , digestion, and feeding behavior. From an evolutionary viewpoint, the major challenge will then logically concern the elucidation of the FLP repertoire of orphan mollusk classes and the way they are functionally related. In this respect, deciphering FLP signaling pathways by characterizing the specific receptors these peptides bind remains another exciting objective.

Published
2014

32. Transcriptomic and peptidomic analysis of protein hydrolysates from the white shrimp (L. vannamei)

Author

Joël Henry, Vincent Fournier, Marie Robert, Benoit Bernay, Gildas Le Corguillé, Céline Zatylny-Gaudin, and Erwan Corre

Subjects
Proteomics, Proteome, Protein Hydrolysates, Size-exclusion chromatography, Molecular Sequence Data, Litopenaeus, Bioengineering, Biology, Mass spectrometry, Applied Microbiology and Biotechnology, Hydrolysate, Mass Spectrometry, Aquaculture, Penaeidae, Sequence Analysis, Protein, Animals, Food science, Amino Acid Sequence, Chromatography, Molecular mass, Bacteria, business.industry, Gene Expression Profiling, Hydrolysis, General Medicine, biology.organism_classification, Shrimp, Anti-Bacterial Agents, Composition (visual arts), business, Peptides, Biotechnology, Chromatography, Liquid
Abstract

An RNAseq approach associated to mass spectrometry was conducted to assess the composition, molecular mass distribution and primary sequence of hydrolytic peptides issued from hydrolysates of white shrimp (Litopenaeus vannamei) by-products. High performance size exclusion chromatography (HPSEC) analyses indicated that 69.2% of the 214-nm-absorbing components had apparent molecular masses below 1000 Da, and 88.3% below 2000 Da. OFFGEL-nLC-MALDI-TOF/TOF and nLC-ESI-MS/MS analyses led to the identification of 808 peptides based on the NCBI EST databank (161,397 entries) completed by the new L. vannamei databank (58,508 entries) that we created from the RNAs of tissues used for hydrolysate production. Whereas most of hydrolytic peptides have a MW below 2000 Da, preliminary investigations of antimicrobial properties revealed three antibacterial fractions that demonstrate functional activities. The abundance of small peptides as well as the biological activities detected could imply very interesting applications for shrimp hydrolysate in the field of aquaculture feeding.

Published
2014

33. Cellular effects of bacterial N-3-Oxo-dodecanoyl-L-Homoserine lactone on the sponge Suberites domuncula (Olivi, 1792): insights into an intimate inter-kingdom dialogue

Author

Gaël Le Pennec, Johan Gardères, Andrés Ritter, Joël Henry, Werner E.G. Müller, Céline Zatylny-Gaudin, Matthias Wiens, Benoit Bernay, Laboratoire de Biotechnologie et Chimie Marines (LBCM), Université de Bretagne Sud (UBS)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS), Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Interactions Cellules Organismes Environnement (ICORE), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU), Departamento de Ecología Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile (UC), Institute for Physiological Chemistry and Pathobiochemistry, Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, and Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)

Subjects
Proteomics, Apoptosis, Pathogenesis, Pathology and Laboratory Medicine, Biochemistry, caspase 7, 4-Butyrolactone, caspase 3, lcsh:Science, Cytoskeleton, caspase like 7 gene, 0303 health sciences, Toll-like receptor, Marine Ecology, toll like receptor, Genomics, proto oncogene, Endocytosis, Cell biology, Suberites domuncula, Cellular Structures and Organelles, alpha actinin, Cell signaling, toll like receptor associated factor 6, Gram negative bacterium, paracrine signaling, Microbiology, 03 medical and health sciences, Genetics, RNA, Messenger, host pathogen interaction, protein expression, two dimensional electrophoresis, Bacteria, 030306 microbiology, Ecology and Environmental Sciences, lcsh:R, Biology and Life Sciences, Computational Biology, Immunity, Innate, carrier protein, Sponge, bacterial membrane, lcsh:Q, immunological tolerance, Suberites, Protein Abundance, suberites domuncula, [SDV]Life Sciences [q-bio], lcsh:Medicine, Molecular Cell Biology, Medicine and Health Sciences, innate immunity, perforin, Multidisciplinary, Ecology, biology, messenger RNA, article, cell communication, Animal Models, matrix assisted laser desorption ionization time of flight mass spectrometry, unclassified drug, Porifera, Host-Pathogen Interactions, cytotoxicity, actin, Transcriptome Analysis, hormone action, Research Article, Symbiotic bacteria, protein bcl 2, Marine Biology, cofilin, Research and Analysis Methods, n (3 oxododecanoyl)homoserine lactone, Microbial Ecology, cognin, Model Organisms, Homoserine, Animals, controlled study, 14. Life underwater, gene, Symbiosis, cell viability, adenosine triphosphatase, 030304 developmental biology, nonhuman, Chemical Ecology, Membrane Proteins, Cell Biology, tumor necrosis factor receptor associated factor 6, Genome Analysis, biology.organism_classification, alpha tubulin, Gene Expression Regulation, Membrane protein, gene expression, caspase like 3 gene, Genome Expression Analysis
Abstract

International audience; Sponges and bacteria have lived together in complex consortia for 700 million years. As filter feeders, sponges prey on bacteria. Nevertheless, some bacteria are associated with sponges in symbiotic relationships. To enable this association, sponges and bacteria are likely to have developed molecular communication systems. These may include molecules such as N-acyl-L-homoserine lactones, produced by Gram-negative bacteria also within sponges. In this study, we examined the role of N-3-oxododecanoyl-L-homoserine lactone (3-oxo-C12-HSL) on the expression of immune and apoptotic genes of the host sponge Suberites domuncula. This molecule seemed to inhibit the sponge innate immune system through a decrease of the expression of genes coding for proteins sensing the bacterial membrane: a Toll-Like Receptor and a Toll-like Receptor Associated Factor 6 and for an anti-bacterial perforin-like molecule. The expression of the pro-apoptotic caspase-like 3/7 gene decreased as well, whereas the level of mRNA of anti-apoptotic genes Bcl-2 Homolog Proteins did not change. Then, we demonstrated the differential expression of proteins in presence of this 3-oxo-C12-HSL using 3D sponge cell cultures. Proteins involved in the first steps of the endocytosis process were highlighted using the 2D electrophoresis protein separation and the MALDI-TOF/TOF protein characterization: α and β subunits of the lysosomal ATPase, a cognin, cofilins-related proteins and cytoskeleton proteins actin, α tubulin and α actinin. The genetic expression of some of these proteins was subsequently followed. We propose that the 3-oxo-C12-HSL may participate in the tolerance of the sponge apoptotic and immune systems towards the presence of bacteria. Besides, the sponge may sense the 3-oxo-C12-HSL as a molecular evidence of the bacterial presence and/or density in order to regulate the populations of symbiotic bacteria in the sponge. This study is the first report of a bacterial secreted molecule acting on sponge cells and regulating the symbiotic relationship.

Published
2014

34. Identification and expression of two oxytocin/vasopressin-related peptides in the cuttlefish Sepia officinalis

Author

Benoit Bernay, Valérie Cornet, Céline Zatylny-Gaudin, Joël Henry, Biologie des mollusques marins et des écosystèmes associés (BioMEA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS), Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, and Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)

Subjects
Signal peptide, Cuttlefish, Central Nervous System, Male, medicine.medical_specialty, Vasopressin, Protein Structure, Sepia, Physiology, Vasopressins, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Neuropeptide, Gene Expression, Neurophysins, Biology, Oxytocin, Biochemistry, 03 medical and health sciences, Cellular and Molecular Neuroscience, 0302 clinical medicine, Endocrinology, Sequence Analysis, Protein, Internal medicine, medicine, Journal Article, Animals, Amino Acid Sequence, Peptide sequence, 030304 developmental biology, 0303 health sciences, Base Sequence, Protein, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sequence Analysis, DNA, DNA, Cell biology, Protein Structure, Tertiary, Female, Sequence Alignment, Sequence Analysis, 030217 neurology & neurosurgery, Tertiary, hormones, hormone substitutes, and hormone antagonists, medicine.drug, Muscle Contraction
Abstract

International audience; Two novel members of the oxytocin/vasopressin superfamily have been identified in the cephalopod Sepia officinalis. Oxytocin/vasopressin gene sequences were cloned by Race PCR. The two precursors we identified exhibit the classical organization of OT/VP superfamily precursors: a signal peptide followed by a nonapeptide and a neurophysin domain. The neurophysin domain is entirely conserved for the cuttlefish precursors, but the nonapeptides and the signal peptides differ. The first nonapeptide, called sepiatocin, is highly homologous to Octopus vulgaris octopressin. The second nonapeptide, called pro-sepiatocin, shows sequence homologies with a Crustacean oxytocin/vasopressin-like peptide identified in Daphnia culex and with a novel form of oxytocin described in New World monkeys. The expression of pro-sepiatocin is restricted to the supraesophageal and subesophageal masses of the brain whereas sepiatocin is expressed in the entire central nervous system. Sepiatocin, as described for octopressin, modulates the contractile activity of several muscles such as penis, oviduct and vena cava muscles; this suggests its involvement in reproduction and blood circulation. Pro-sepiatocin is released in the hemolymph; it is a neurohormone able to target numerous peripheral organs.

Published
2013

35. Characterization of GnRH-related peptides from the Pacific oyster Crassostrea gigas

Author

Pierre Boudry, Franck Rodet, Céline Zatylny-Gaudin, Pascal Favrel, Laetitia Bigot, Benoit Bernay, Biologie des mollusques marins et des écosystèmes associés ( BioMEA ), Université de Caen Normandie ( UNICAEN ), Normandie Université ( NU ) -Normandie Université ( NU ) -Institut Français de Recherche pour l'Exploitation de la Mer ( IFREMER ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Spectrométrie de Masse Biologique Fondamentale et Appliquée, Université de Lille, Sciences et Technologies, Laboratoire des Sciences de l'Environnement Marin (LEMAR) ( LEMAR ), Institut de Recherche pour le Développement ( IRD ) -Institut Français de Recherche pour l'Exploitation de la Mer ( IFREMER ) -Université de Brest ( UBO ) -Institut Universitaire Européen de la Mer ( IUEM ), Institut de Recherche pour le Développement ( IRD ) -Université de Brest ( UBO ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université de Brest ( UBO ) -Centre National de la Recherche Scientifique ( CNRS ) -Centre National de la Recherche Scientifique ( CNRS ), Biologie des mollusques marins et des écosystèmes associés (BioMEA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS), Laboratoire des Sciences de l'Environnement Marin (LEMAR) (LEMAR), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), and Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, Oyster, Insecta, Mollusk, Physiology, Molecular Sequence Data, Neuropeptide, Zoology, Gene Expression, Gonadotropin-releasing hormone, Neuroendocrinology, Biochemistry, Mass Spectrometry, Gonadotropin-Releasing Hormone, [ SDE ] Environmental Sciences, 03 medical and health sciences, Cellular and Molecular Neuroscience, 0302 clinical medicine, Endocrinology, biology.animal, Animals, Amino Acid Sequence, Crassostrea, Gene, Peptide sequence, Chromatography, High Pressure Liquid, Phylogeny, 030304 developmental biology, 0303 health sciences, biology, Sequence Homology, Amino Acid, Ecology, Tissue Extracts, Reproduction, Bivalve, Pacific oyster, 16. Peace & justice, biology.organism_classification, Ganglia, Invertebrate, Starvation, GnRH, Vertebrates, [SDE]Environmental Sciences, Female, 030217 neurology & neurosurgery
Abstract

Gonadotropin-releasing hormone (GnRH), a key neuropeptide regulating reproduction in vertebrates has now been characterized in a number of non-vertebrate species. Despite the demonstration of its ancestral origin, the structure and the function of this family of peptides remain poorly known in species as distant as lophotrochozoans. In this study, two GnRH-related peptides (Cg-GnRH-a and CgGnRH-G) were characterized by mass spectrometry from extracts of the visceral ganglia of the Pacific oyster Crassostrea gigas. These peptides showed a high degree of sequence identity with GnRHs of other mollusks and annelids and to a lesser extent with those of vertebrates or with AKH and corazonins of insects. Both the mature peptides and the transcript encoding the precursor protein were exclusively expressed in the visceral ganglia. Significant differences in transcriptional activity of Cg-GnRH encoding gene were recorded in the ganglia along the reproductive cycle and according to trophic conditions with a higher level in fed animals compared to starved animals. This suggests the involvement of Cg-GnRHs as synchronizers of nutritional status with energy requirements during reproduction in oyster. Evidence for a role of Cg-GnRHs as neuroregulators and as neuroendocrine factors in bivalve is discussed. (C) 2012 Elsevier Inc. All rights reserved.

Published
2012

36. From a marine neuropeptide to antimicrobial pseudopeptides containing aza-β(3)-amino acids: structure and activity

Author

Joël Henry, Michèle Baudy-Floc’h, Arnaud Bondon, Baptiste Legrand, Céline Zatylny-Gaudin, Mathieu Laurencin, Emilie Duval, Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Structures et interactions moléculaires, Institut de Génétique et Développement de Rennes (IGDR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Physiologie et Ecophysiologie des Mollusques Marins (PE2M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS), We thank SERB Laboratories and Region Bretagne for their financial support., Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and De Villemeur, Hervé

Subjects
Models, Molecular, Circular dichroism, MESH: Amino Acids, Magnetic Resonance Spectroscopy, Molecular Conformation, MESH: Rabbits, MESH: Cricetinae, Peptide, MESH: Neuropeptides, 01 natural sciences, MESH: Gram-Positive Bacteria, MESH: Circular Dichroism, MESH: Structure-Activity Relationship, MESH: Cricetulus, Cricetinae, MESH: Gram-Negative Bacteria, Drug Discovery, MESH: Animals, Amino Acids, chemistry.chemical_classification, MESH: Microbial Sensitivity Tests, Oligopeptide, Chemistry, Circular Dichroism, MESH: Antimicrobial Cationic Peptides, Stereoisomerism, Nuclear magnetic resonance spectroscopy, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, MESH: Hemolysis, Amino acid, Anti-Bacterial Agents, [SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, MESH: Oligopeptides, Molecular Medicine, Rabbits, Oligopeptides, MESH: Models, Molecular, Stereochemistry, [SDV.BBM.BP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, CHO Cells, Microbial Sensitivity Tests, 010402 general chemistry, Gram-Positive Bacteria, Hemolysis, Article, Turn (biochemistry), Structure-Activity Relationship, Cricetulus, MESH: CHO Cells, MESH: Anti-Bacterial Agents, Gram-Negative Bacteria, Structure–activity relationship, Animals, MESH: Molecular Conformation, Aza Compounds, MESH: Magnetic Resonance Spectroscopy, 010405 organic chemistry, Neuropeptides, MESH: Stereoisomerism, 0104 chemical sciences, MESH: Aza Compounds, Antimicrobial Cationic Peptides
Abstract

Incorporation of aza-β3-amino acids into endogenous neuropeptide from mollusks (ALSGDAFLRF-NH2) with weak antimicrobial activities allows us to design new AMPs sequences. We find that, depending on the nature of the substitution, these could result either in inactive pseudopeptides or in a drastic enhancement of the antimicrobial activity without high cytotoxicity resulted. Structural studies perform by NMR and circular dichroism on the pseudopeptides show the impact of aza-β3-amino acids on the peptide structures. We obtain the first three-dimensional structures of pseudopeptides containing aza-β3-amino acids in aqueous micellar SDS and demonstrate that hydrazino turn can be formed in aqueous solution. Overall, these results demonstrate the ability to modulate AMPs activities through structural modifications induced by the nature and the position of these amino acid analogs in the peptide sequences.

Published
2012

37. A complex set of sex pheromones identified in the cuttlefish Sepia officinalis

Author

Joël Henry, Benoit Bernay, Michèle Baudy-Floc’h, Jérôme Leprince, Jérémy Enault, Céline Zatylny-Gaudin, Benjamin Lefranc, Biologie des mollusques marins et des écosystèmes associés (BioMEA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS), Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-High-tech Research Infrastructures for Life Sciences (HeRacLeS), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), This work was supported by a Ph.D. grant of the 'Comité Régional de Basse-Normandy'., Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Brébion, Alice, and Lefranc, Benjamin

Subjects
Cuttlefish, Sexual Reproduction, Anatomy and Physiology, lcsh:Medicine, Bioinformatics, Biochemistry, Mass Spectrometry, Pheromones, 0302 clinical medicine, Sepia, Mating, Sex Attractants, lcsh:Science, Peptide sequence, Animal Management, Genetics, Expressed Sequence Tags, 0303 health sciences, Expressed sequence tag, [SDV.BDLR.RS] Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, Multidisciplinary, Animal Behavior, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Reproduction, Decapodiformes, Sex pheromone, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Female, Seasons, Research Article, Sexual Behavior, Molecular Sequence Data, Endocrine System, Marine Biology, Biology, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, 03 medical and health sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Animals, Amino Acid Sequence, 030304 developmental biology, Gene Library, Endocrine Physiology, cDNA library, Gene Expression Profiling, lcsh:R, Reproductive System, Proteins, Fisheries Science, Sequence Analysis, DNA, [CHIM.ORGA] Chemical Sciences/Organic chemistry, Hormones, Regulatory Proteins, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, [SDV.BA.ZI] Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, lcsh:Q, Animal Migration, Veterinary Science, Peptides, 030217 neurology & neurosurgery
Abstract

International audience; BACKGROUND: The cephalopod mollusk Sepia officinalis can be considered as a relevant model for studying reproduction strategies associated to seasonal migrations. Using transcriptomic and peptidomic approaches, we aim to identify peptide sex pheromones that are thought to induce the aggregation of mature cuttlefish in their egg-laying areas. RESULTS: To facilitate the identification of sex pheromones, 576 5'-expressed sequence tags (ESTs) were sequenced from a single cDNA library generated from accessory sex glands of female cuttlefish. Our analysis yielded 223 unique sequences composed of 186 singletons and 37 contigs. Three major redundant ESTs called SPα, SPα' and SPβ were identified as good candidates for putative sex pheromone transcripts and are part of the 87 unique sequences classified as unknown. The alignment of translated SPα and SPα' revealed a high level of conservation, with 98.4% identity. Translation led to a 248-amino acid precursor containing six peptides with multiple putative disulfide bonds. The alignment of SPα-α' with SPβ revealed a partial structural conservation, with 37.3% identity. Translation of SPβ led to a 252-amino acid precursor containing five peptides. The occurrence of a signal peptide on SPα, SPα' and SPβ showed that the peptides were secreted. RT-PCR and mass spectrometry analyses revealed a co-localization of transcripts and expression products in the oviduct gland. Preliminary in vitro experiments performed on gills and penises revealed target organs involved in mating and ventilation. CONCLUSIONS: The analysis of the accessory sex gland transcriptome of Sepia officinalis led to the identification of peptidic sex pheromones. Although preliminary functional tests suggested the involvement of the α3 and β2 peptides in ventilation and mating stimulation, further functional investigations will make it possible to identify the complete set of biological activities expected from waterborne pheromones.

Published
2012

39. Structure and mechanism of action of a de novo antimicrobial detergent-like peptide

Author

Céline Zatylny-Gaudin, Liza Mouret, Joe Sarkis, Michèle Baudy-Floc’h, Murielle Collen, Arnaud Bondon, Joël Henry, Baptiste Legrand, Véronique Vié, Mathieu Laurencin, Emilie Duval, Jean-François Hubert, Interactions cellulaires et moléculaires (ICM), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de Physique de Rennes (IPR), Région Bretagne and SERB pharmaceutical company, Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), and Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)

Subjects
Models, Molecular, Circular dichroism, MESH: Hydrogen-Ion Concentration, Magnetic Resonance Spectroscopy, Protein Conformation, Molecular Conformation, Peptide, Microscopy, Atomic Force, Biochemistry, Micelle, MESH: Circular Dichroism, chemistry.chemical_compound, Protein structure, MESH: Protein Conformation, Anti-Infective Agents, chemistry.chemical_classification, MESH: Microscopy, Atomic Force, 0303 health sciences, Liposome, MESH: Peptides, Vesicle, Bilayer, 030302 biochemistry & molecular biology, Self-assembly, Hydrogen-Ion Concentration, Fluoresceins, Lipids, MESH: Microscopy, Electron, Transmission, Protons, Antimicrobial peptide, MESH: Models, Molecular, Detergent, Stereochemistry, MESH: Fluoresceins, MESH: Anti-Infective Agents, Detergents, Biophysics, Phospholipid, 03 medical and health sciences, Microscopy, Electron, Transmission, MESH: Water, 030304 developmental biology, MESH: Molecular Conformation, MESH: Magnetic Resonance Spectroscopy, Water, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell Biology, MESH: Lipids, NMR, chemistry, MESH: Protons, Peptides, MESH: Detergents
Abstract

International audience; The K4 peptide (KKKKPLFGLFFGLF) was recently demonstrated to display good antimicrobial activities against various bacterial strains and thus represents a candidate for the treatment of multiple-drug resistant infections. In this study, we use various techniques to study K4 behaviour in different media: water, solutions of detergent micelles, phospholipid monolayers and suspension of phospholipid vesicles. First, self-assembly of the peptide in water is observed, leading to the formation of spherical objects around 10nm in diameter. The addition of micelles induces partial peptide folding to an extent depending on the charge of the detergent headgroups. The NMR structure of the peptide in the presence of SDS displays a helical character of the hydrophobic moiety, whereas only partial folding is observed in DPC micelles. This peptide is able to destabilize the organization of monolayer membranes or bilayer liposomes composed of anionic lipids. When added on small unilamellar vesicles it generates larger objects attributed to mixed lipid-peptide vesicles and aggregated vesicles. The absence of calcein leakage from liposomes, when adding K4, underlines the original mechanism of this linear amphipathic peptide. Our results emphasize the importance of the electrostatic effect for K4 folding and lipid destabilization leading to the microorganisms' death with a high selectivity for the eukaryotic cells at the MIC. Interestingly, the micrographs obtained by electronic microscopy after addition of peptide on bacteria are also consistent with the formation of mixed lipid-peptide objects. Overall, this work supports a detergent-like mechanism for the antimicrobial activity of this peptide.

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40. Study of the immune system in the cuttlefish Sepia officinalis : potential uses in aquaculture

Author

Benoist, Louis, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Normandie Université, and Céline Zatylny-Gaudin

Subjects
Mucus, Haemocytes, Haematopoiesis, White body, [SDV.BA]Life Sciences [q-bio]/Animal biology, Omic, Immunity, Corps blanc, Microbiome, Skin
Abstract

Cephalopods such as the cuttlefish Sepia officinalis, despite their short lifespan, have been found in the oceans for millions of years. In these atypical animals, few pathologies have been observed, revealing the presence of an effective but little studied immune system based on innate processes. The study of the cuttlefish's immune system has been carried out on the white body, a haematopoietic organ; on the circulating cells, the haemocytes; and on the skin, the first barrier with the external environment. At the white body level, the transcriptomic and proteomic study highlighted the presence of factors linked to haematopoiesis, including members of the JAK-STAT signalling pathway. Immune factors have also been identified, revealing a possible multifunctionality of the white body. The immune response to Vibrio splendidus could be apprehended from a comparative transcriptomic analysis of haemocytes. However, as the latter did not allow the clear identification of antimicrobial peptides, an original in silico analysis was developed to select five candidate peptides, three of which revealed a targeted antibacterial activity against bacteria of the Vibrio genus. Finally, a study of the skin and its mucus was initiated. This study using -omic approaches enabled the identification of factors related to pathogen recognition and immune response. In addition, twelve strains were isolated and identified at the level of the skin microbiome. All these results represent a major contribution concerning the immune system in cuttlefish, making it possible to initiate functional studies during an infection or at the end of life. These studies would make it possible to understand the mode of action of the identified immune factors, the involvement of each entity in the immune response or in the establishment and maintenance of the microbiome.; Les Céphalopodes comme la seiche Sepia officinalis, malgré une durée de vie courte, sont retrouvés dans les océans depuis des millions d’années. Chez ces animaux atypiques, peu de pathologies ont été observées révélant la présence d’un système immunitaire efficace mais peu étudié qui repose sur des processus innés. L’étude du système immunitaire de la seiche a été menée au niveau du corps blanc, organe hématopoïétique ; des cellules circulantes, les hémocytes et au niveau de la peau, première barrière avec le milieu extérieur. Au niveau du corps blanc, l’étude transcriptomique et protéomique a mis en évidence la présence de facteurs en lien avec l’hématopoïèse dont des membres de la voie de signalisation JAK-STAT. Des facteurs immunitaires ont également été identifiés révélant une possible multifonctionnalité du corps blanc. La réponse immunitaire face à Vibrio splendidus a pu être appréhendées à partir d’une analyse transcriptomique comparative sur les hémocytes. Toutefois cette dernière n’ayant pas permis d’identifier clairement des peptides antimicrobiens, une analyse in silico originale a été développée permettant de sélectionner cinq peptides candidats dont trois ont révélé une activité antibactérienne ciblée contre des bactéries du genre Vibrio. Enfin, une étude au niveau de la peau et de son mucus a été initiée. Cette étude par des approches -omiques a permis l’identification de facteurs en lien avec la reconnaissance des pathogènes et la réponse immunitaire. Par ailleurs, douze souches ont pu être isolées et identifiées au niveau du microbiome cutané. L’ensemble de ces résultats représente un apport majeur concernant le système immunitaire chez la seiche permettant d’initier des études fonctionnelles lors d’une infection ou en fin de vie. Ces études permettraient de comprendre le mode d’action des facteurs immunitaires identifiés, l’implication de chaque entité dans la réponse immunitaire ou dans la mise en place et la maintenance du microbiome.

Published
2020

41. Identification de peptides antibactériens d'origine marine : Amélioration de la qualité et de la survie du naissain d'huître

Author

Houyvet, Baptiste, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Normandie Université, and Céline Zatylny-Gaudin

Subjects
Pterois volitans, Transcriptomique, Peptides antibactériens, Écloserie, Databases, [SDV.BA]Life Sciences [q-bio]/Animal biology, Peptidomique, Peptidomics, Sepia officinalis, Trancriptomics, Hatchery, Antibacterial peptides
Abstract

The first larval stages of oyster (Magallana gigas) are key steps in the smooth running of the zootechnical course and inthe sustainability of hatcheries, where mortality levels can be high. That is why we searched for new antimicrobialpeptides (AMPs) on two marine organisms, i.e. lionfish (Pterois volitans), which is invasive in the Caribbean Sea, and thecommon cuttlefish (Sepia officinalis), which is present in French oyster production areas. The search for AMPs wascarried out preferentially from de novo transcriptomes from these two animals. In lionfish, BLAST analyses allowed forthe identification of 7 transcripts encoding AMPs. Four of them shared strong sequence homology (> 90% identity) withAMPs rich in cysteines and close to hepcidin, LEAP-2, NK-lysin and b-defensin identified in other fish. The other 3transcripts, annotated pteroicidins A, B and C, coded for piscidin-related peptides. The presence of b-defensin andpteroicidin a encoded by pteroicidin A was confirmed in lionfish skin extracts by mass spectrometry. An in-depth studywas conducted on two amide and non-amide forms of pteroicidin a, as well as on several peptides of different sizesderived from pteroicidins B and C. The results highlighted a relationship between structure, amidation, and theantibacterial and hemolytic activities of these different pteroicidins. On the other hand, no AMP was highlighted in theSepia officinalis model using conventional approaches coupling purification and antibacterial tests or BLAST approaches.We therefore developed a more original approach that relies on the design of peptides starting from the transcriptome.Starting from 811 small cysteine-free peptides from the APD database, we determined recurring criteria for charge,hydrophobicity, and amino acid composition. Based on these criteria and on CAMP prediction tools, twelve peptides weresynthesized. Five of them revealed a broad spectrum of antibacterial activities. Non-hemolytic antibacterial peptidesderived from cuttlefish were transferred to the hatchery. This transfer was optimized thanks to a preliminary study onthe de novo K4 peptide, which is particularly active on vibrios. The study highlighted the importance of antibacterialpeptide safety on the various links of the trophic chain including oyster, and the importance of the targeted stage ofdevelopment. In addition, we addressed the fate of antibacterial peptides to ensure their biodegradability. Altogether,this work not only helped to identify new AMPs but also to provide the first data on the potential use of these peptidesas an alternative to antibiotics.; Les premiers stades larvaires chez l’huitre creuse, nommée Magallana gigas, constituent une étape clé du bondéroulement du parcours zootechnique et également pour la pérennisation de la production en écloserie. Dans l’objectifde réduire les mortalités observées en écloserie, nous avons recherché de nouveaux peptides antimicrobiens. Larecherche de ces PAM a été réalisée à partir de deux organismes marins, le poisson-lion invasif en mer des caraïbes,Pterois volitans, et la seiche commune présente sur les zones ostréicoles, Sepia officinalis. La recherche de PAM a étéréalisée préférentiellement à partir de transcriptomes de novo obtenus chez ces deux animaux. Chez le poisson lion, àpartir de BLAST, 7 transcrits codant pour des PAM ont été identifiés. Quatre de ces PAM partagent de fortes homologiesde séquences (>90% d’identité) avec des PAM riches en cystéines proches de l’hepcidine, la LEAP-2, la NK-lysine et la bdéfensineidentifiées chez d’autres poissons. Les 3 autres transcrits annotés pteroicidines A, B et C codent pour despeptides apparentés aux piscidines. La présence de la b-défensine et de la pteroicidine a codée par la pteroicidine A a puêtre confirmée dans les extrait de peau du poisson lion par spectrométrie de masse. Une étude approfondie a été menéesur deux formes amidée et non amidée de la ptéroicidine a ainsi que sur plusieurs peptides de différentes tailles issusdes pteroicidines B et C. Les résultats ont permis de mettre en évidence une relation entre la structure, l’amidation et lesactivités antibactériennes et hémolytiques de ces différentes ptéroicidines. Sur le modèle Sepia officinalis, par lesapproches classiques couplant la purification et les tests antibactériens ou par des approches utilisant les BlAST, aucunPAM n’a été mis en évidence. Nous avons donc développé une approche plus originale qui repose sur le « design » depeptides à partir du transcriptome. A partir de 811 petits peptides sans cystéines issus de la base de données APD, nousavons déterminé des critères récurrents concernant la charge, l’hydrophobicité et la composition en acides aminés. Surla base de ces critères et en s’appuyant sur les outils de prédiction de CAMP, douze peptides ont fait l’objet d’une synthèse.Cinq de ces peptides ont révélé un large spectre d’activités antibactériennes. Les peptides antibactériens issus de la seicheayant une activité non hémolytique ont fait l’objet d’un transfert en écloserie. Ce transfert a été optimisé à partir d’uneétude préliminaire sur le peptide de novo K4, particulièrement actif sur les vibrios. Cette étude a mis en évidencel’importance de l’innocuité du peptide antibactérien sur les différents maillons de la chaine trophique, notamment del’huitre, et sur l’importance du stade de développement ciblé. Par ailleurs, nous nous sommes intéressés au devenir despeptides antibactériens de manière à s’assurer de leur biodégradabilité. L’ensemble de ces travaux a permis nonseulement d’identifier de nouveaux PAMs mais également d’apporter les premières données portant sur le potentiel del’utilisation de ces peptides comme alternative aux antibiotiques.

Published
2018

42. A single regulatory gene is sufficient to alter Vibrio aestuarianus pathogenicity in oysters

Author

Goudenege, David, Travers, Marie-agnes, Lemire, Astrid, Petton, Bruno, Haffner, Philippe, Labreuche, Yannick, Tourbiez, Delphine, Mangenot, Sophie, Calteau, Alexandra, Mazel, Didier, Nicolas, Jean-louis, Jacq, Annick, Le Roux, Frederique, Unité Physiologie Fonctionnelle des Organismes marins, Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Institut Français de Recherche pour l'Exploitation de la Mer - Brest (IFREMER Centre de Bretagne), Laboratoire de Génétique et Pathologie des Mollusques Marins, 17390 La Tremblade, France. (LGPMM), Santé, Génétique et Microbiologie des Mollusques (IFREMER SG2M), Institut Français de Recherche pour l'Exploitation de la Mer - Atlantique (IFREMER Atlantique), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer - Atlantique (IFREMER Atlantique), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire des Sciences de l'Environnement Marin (LEMAR) (LEMAR), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Génomique d'Evry (IG), Institut de Biologie François JACOB (JACOB), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Analyse Bio-Informatique pour la Génomique et le Métabolisme (LABGeM), Génomique métabolique (UMR 8030), Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université d'Évry-Val-d'Essonne (UEVE)-Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université d'Évry-Val-d'Essonne (UEVE), Plasticité du Génome Bactérien - Bacterial Genome Plasticity (PGB), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut de génétique et microbiologie [Orsay] (IGM), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), The present study has been supported by the DPMA (Convention DPMA 2013- IFREMER 12/1210320/NYF), the ANR Blanc (11-BSV7-023-01 «VIBRIOGEN», D. G. funding), the EMBRC France (A. L. funding) and the ANR Bioadapt (13-ADAP-0007-01 «OPOPOP»)., We warmly thank Dr Maurizio Labbate (University of Technology, Sydney, Australia), Dr Maxime Bruto (Station Biologique de Roscoff) and Dr Marianne Alunno-Bruscia (Ifremer Argenton) for critically reading the manuscript. We also thank Pr Joël Henry and Dr Céline Zatylny-Gaudin (Université de Caen Basse-Normandie, Institut de Biologie Fondamentale et Appliquée) for kindly performing mass spectrometry analyses. We thank Dr Carla Pruzzo (University of Genoa, Italy) and Dr Ana Roque (IRTA, Spain) for providing U_17 and KB19 strains. We acknowledge the staff of the station Ifremer Argenton, La Tremblade, Bouin, particularly Max Nourry, the ABIMS (SBR Roscoff) and LABGeM (Evry) platforms for technical support., ANR-11-BSV7-0023,VIBRIOGEN,Les vibrios pathogènes d'invertébrés marins : un nouveau regard sur la virulence et les réservoirs de gènes permettant l'adaptation à la niche écologique(2011), ANR-13-ADAP-0007,OPOPOP,Emergence de pathogènes opportunistes d'huîtres dans des populations naturelles de Vibrio(2013), Physiologie Fonctionnelle des Organismes Marins (PFOM), Laboratoire de Génétique et Pathologie des Mollusques Marins (LGPMM), Santé, Génétique et Microbiologie des Mollusques (SGMM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Saclay-Institut de Biologie François JACOB (JACOB), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS)-Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Français de Recherche pour l'Exploitation de la Mer ( IFREMER ), Laboratoire de Biologie Intégrative des Modèles Marins ( LBI2M ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Génomique d'Evry ( IG ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay, Génomique métabolique ( UMR 8030 ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université d'Évry-Val-d'Essonne ( UEVE ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ), Plasticité du Génome Bactérien, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Institut de génétique et microbiologie [Orsay] ( IGM ), Université Paris-Sud - Paris 11 ( UP11 ) -Centre National de la Recherche Scientifique ( CNRS ), ANR-11-BSV7-0023,VIBRIOGEN,Les vibrios pathogènes d'invertébrés marins : un nouveau regard sur la virulence et les réservoirs de gènes permettant l'adaptation à la niche écologique ( 2011 ), ANR-13-ADAP-0007,OPOPOP,Emergence de pathogènes opportunistes d'huîtres dans des populations naturelles de Vibrio ( 2013 ), Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS)-Université de Brest (UBO), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université d'Évry-Val-d'Essonne (UEVE)-Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université d'Évry-Val-d'Essonne (UEVE)

Subjects
MESH: Genes, Regulator, [SDV]Life Sciences [q-bio], CHOLERAE HEMAGGLUTININ PROTEASE, PACIFIC OYSTERS, MESH : Protein Kinases, MESH: Virulence, MESH: Ostreidae, MESH : Frameshift Mutation, MESH: Vibrio, Genes, Regulator, Animals, LETHALITY, MESH: Animals, MESH : Genes, Regulator, METALLOPROTEASE, MESH : France, Frameshift Mutation, MESH: Phylogeny, MORTALITIES, MESH: Protein Kinases, Phylogeny, Vibrio, CRASSOSTREA-GIGAS, MESH : Ostreidae, [ SDV ] Life Sciences [q-bio], CONSTRUCTION, ACL, MESH: Genomics, MESH : Genomics, ALGORITHMS, MESH : Virulence, MESH: Frameshift Mutation, MESH : Phylogeny, Genomics, Ostreidae, FAMILY, MESH: France, VIRULENCE, MESH : Animals, France, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, Protein Kinases, MESH : Vibrio
Abstract

International audience; Oyster diseases caused by pathogenic vibrios pose a major challenge to the sustainability of oyster farming. In France, since 2012 a disease affecting specifically adult oysters has been associated with the presence of Vibrio aestuarianus. Here, by combining genome comparison, phylogenetic analyses and high-throughput infections of strains isolated before or during the recent outbreaks, we show that virulent strains cluster into two V. aestuarianus lineages independently of the sampling dates. The bacterial lethal dose was not different between strains isolated before or after 2012. Hence, the emergence of a new highly virulent clonal strain is unlikely. Each lineage comprises nearly identical strains, the majority of them being virulent, suggesting that within these phylogenetically coherent virulent lineages a few strains have lost their pathogenicity. Comparative genomics allowed the identification of a single frameshift in a non-virulent strain. This mutation affects the varS gene that codes for a signal transduction histidine-protein kinase. Genetic analyses confirmed that varS is necessary for infection of oysters and for a secreted metalloprotease expression. For the first time in a Vibrio species, we show here that VarS is a key factor of pathogenicity.

Published
2015

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