RESUMEN
Long-chain n-3 PUFA (LC n-3 PUFA) prevent, in rodents, insulin resistance (IR) induced by a high-fat and/or fructose diet but not IR induced by glucocorticoids. In humans, contrasting effects have also been reported. We investigated their effects on insulin sensitivity, feed intake (FI) and body weight gain in genetically insulin resistant male obese (fa/fa) Zucker (ZO) rats during the development of obesity. ZO rats were fed a diet supplemented with 7 % fish oil (FO) + 1 % corn oil (CO) (wt/wt) (ZOFO), while the control group was fed a diet containing 8 % fat from CO (wt/wt) (ZOCO). Male lean Zucker (ZL) rats fed either FO (ZLFO) or CO (ZLCO) diet were used as controls. FO was a marine-derived TAG oil containing EPA 90 mg/g + DHA 430 mg/g. During an oral glucose tolerance test, glucose tolerance remained unaltered by FO while insulin response was reduced in ZOFO only. Liver insulin sensitivity (euglycaemic-hyperinsulinaemic clamp + 2 deoxyglucose) was improved in ZOFO rats, linked to changes in phosphoenolpyruvate carboxykinase expression, activity and glucose-6-phosphatase activity. FI in response to intra-carotid insulin/glucose infusion was decreased similarly in ZOFO and ZOCO. Hypothalamic ceramides levels were lower in ZOFO than in ZOCO. Our study demonstrates that LC n-3 PUFA can minimise weight gain, possibly by alleviating hypothalamic lipotoxicity, and liver IR in genetically obese Zucker rats.
Asunto(s)
Ácidos Grasos Omega-3 , Resistencia a la Insulina , Humanos , Masculino , Ratas , Animales , Resistencia a la Insulina/fisiología , Aceites de Pescado/farmacología , Ratas Zucker , Glucemia/metabolismo , Insulina/metabolismo , Obesidad/metabolismo , Glucosa/farmacología , Ingestión de Alimentos , Aumento de Peso , Ácidos Grasos Insaturados/farmacología , Aceite de Maíz/farmacología , Ácidos Grasos Omega-3/farmacologíaRESUMEN
The Manila clam (Ruditapes philippinarum) is the second most exploited bivalve in the world but remains threatened by diseases and global changes. Their associated microbiota play a key role in their fitness and acclimation capacities. This study aimed at better understanding the behavior of clam digestive glands and extrapallial fluids microbiota at small, but contrasting spatial and temporal scales. Results showed that environmental variations impacted clam microbiota differently according to the considered tissue. Each clam tissue presented its own microbiota and showed different dynamics according to the intertidal position and sampling period. Extrapallial fluids microbiota was modified more rapidly than digestive glands microbiota, for clams placed on the upper and lower intertidal position, respectively. Clam tissues could be considered as different microhabitats for bacteria as they presented different responses to small-scale temporal and spatial variabilities in natural conditions. These differences underlined a more stringent environmental filter capacity of the digestive glands.
Asunto(s)
Bivalvos , Microbiota , Animales , Bivalvos/microbiologíaRESUMEN
Among all the environmental factors, seawater temperature plays a decisive role in triggering marine diseases. Like fever in vertebrates, high seawater temperature could modulate the host response to the pathogens in ectothermic animals. In France, massive mortality of Pacific oysters Crassostrea gigas caused by the ostreid herpesvirus 1 (OsHV-1) is markedly reduced when temperatures exceed 24°C in the field. In the present study we assess how high temperature influences the host response to the pathogen by comparing transcriptomes (RNA-sequencing) during the course of experimental infection at 21°C (reference) and 29°C. We show that high temperature induced host physiological processes that are unfavorable to the viral infection. Temperature influenced the expression of transcripts related to the immune process and increased the transcription of genes related to apoptotic process, synaptic signaling, and protein processes at 29°C. Concomitantly, the expression of genes associated to catabolism, metabolites transport, macromolecules synthesis and cell growth remained low since the first stage of infection at 29°C. Moreover, viral entry into the host might have been limited at 29°C by changes in extracellular matrix composition and protein abundance. Overall, these results provide new insights into how environmental factors modulate the host-pathogen interactions.
RESUMEN
Of all environmental factors, seawater temperature plays a decisive role in triggering marine diseases. Like fever in vertebrates, high seawater temperature could modulate the host response to pathogens in ectothermic animals. In France, massive mortality of Pacific oysters, Crassostrea gigas, caused by the ostreid herpesvirus 1 (OsHV-1) is markedly reduced when temperatures exceed 24°C in the field. In the present study we assess how high temperature influences the host response to the pathogen by comparing transcriptomes (RNA sequencing) during the course of experimental infection at 21°C (reference) and 29°C. We show that high temperature induced host physiological processes that are unfavorable to the viral infection. Temperature influenced the expression of transcripts related to the immune process and increased the transcription of genes related to the apoptotic process, synaptic signaling and protein processes at 29°C. Concomitantly, the expression of genes associated with catabolism, metabolite transport, macromolecule synthesis and cell growth remained low from the first stage of infection at 29°C. Moreover, viral entry into the host might have been limited at 29°C by changes in extracellular matrix composition and protein abundance. Overall, these results provide new insights into how environmental factors modulate host-pathogen interactions.
Asunto(s)
Crassostrea , Herpesviridae , Animales , Crassostrea/genética , Francia , Herpesviridae/genética , Temperatura , TranscriptomaRESUMEN
Plastics are persistent synthetic polymers that accumulate as waste in the marine environment. Microplastic (MP) particles are derived from the breakdown of larger debris or can enter the environment as microscopic fragments. Because filter-feeder organisms ingest MP while feeding, they are likely to be impacted by MP pollution. To assess the impact of polystyrene microspheres (micro-PS) on the physiology of the Pacific oyster, adult oysters were experimentally exposed to virgin micro-PS (2 and 6 µm in diameter; 0.023 mg·L(-1)) for 2 mo during a reproductive cycle. Effects were investigated on ecophysiological parameters; cellular, transcriptomic, and proteomic responses; fecundity; and offspring development. Oysters preferentially ingested the 6-µm micro-PS over the 2-µm-diameter particles. Consumption of microalgae and absorption efficiency were significantly higher in exposed oysters, suggesting compensatory and physical effects on both digestive parameters. After 2 mo, exposed oysters had significant decreases in oocyte number (-38%), diameter (-5%), and sperm velocity (-23%). The D-larval yield and larval development of offspring derived from exposed parents decreased by 41% and 18%, respectively, compared with control offspring. Dynamic energy budget modeling, supported by transcriptomic profiles, suggested a significant shift of energy allocation from reproduction to structural growth, and elevated maintenance costs in exposed oysters, which is thought to be caused by interference with energy uptake. Molecular signatures of endocrine disruption were also revealed, but no endocrine disruptors were found in the biological samples. This study provides evidence that micro-PS cause feeding modifications and reproductive disruption in oysters, with significant impacts on offspring.
Asunto(s)
Ostreidae/fisiología , Plásticos/farmacología , Poliestirenos/farmacología , Reproducción/efectos de los fármacos , Animales , Ostreidae/genética , Ostreidae/metabolismo , Proteoma , TranscriptomaRESUMEN
Temperature triggers marine diseases by changing host susceptibility and pathogen virulence. Oyster mortalities associated with the Ostreid herpesvirus type 1 (OsHV-1) have occurred seasonally in Europe when the seawater temperature range reaches 16-24⯰C. Here we assess how temperature modulates oyster susceptibility to OsHV-1 and pathogen virulence. Oysters were injected with OsHV-1 suspension incubated at 21⯰C, 26⯰C and 29⯰C and were placed in cohabitation with healthy oysters (recipients) at these three temperatures according to a fractional factorial design. Survival was followed for 14â¯d and recipients were sampled for OsHV-1 DNA quantification and viral gene expression. The oysters were all subsequently placed at 21⯰C to evaluate the potential for virus reactivation, before being transferred to oyster farms to evaluate their long-term susceptibility to the disease. Survival of recipients at 29⯰C (86%) was higher than at 21⯰C (52%) and 26⯰C (43%). High temperature (29⯰C) decreased the susceptibility of oysters to OsHV-1 without altering virus infectivity and virulence. At 26⯰C, the virulence of OsHV-1 was enhanced. Differences in survival persisted when the recipients were all placed at 21⯰C, suggesting that OsHV-1 did not reactivate. Additional oyster mortality followed the field transfer, but the overall survival of oysters infected at 29⯰C remained higher.
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Crassostrea/inmunología , Crassostrea/virología , Virus ADN/patogenicidad , Susceptibilidad a Enfermedades , Temperatura , Animales , Virus ADN/genética , ADN Viral/análisis , Femenino , Expresión Génica , Infecciones por Herpesviridae/veterinaria , Masculino , VirulenciaRESUMEN
Here, we assess the physiological effects induced by environmental concentrations of pesticides in Pacific oyster Crassostrea gigas. Oysters were exposed for 14 d to trace levels of metconazole (0.2 and 2 µg/L), isoproturon (0.1 and 1 µg/L), or both in a mixture (0.2 and 0.1 µg/L, respectively). Exposure to trace levels of pesticides had no effect on the filtration rate, growth, and energy reserves of oysters. However, oysters exposed to metconazole and isoproturon showed an overactivation of the sensing-kinase AMP-activated protein kinase α (AMPKα), a key enzyme involved in energy metabolism and more particularly glycolysis. In the meantime, these exposed oysters showed a decrease in hexokinase and pyruvate kinase activities, whereas 2-DE proteomic revealed that fructose-1,6-bisphosphatase (F-1,6-BP), a key enzyme of gluconeogenesis, was up-regulated. Activities of antioxidant enzymes were higher in oysters exposed to the highest pesticide concentrations. Both pesticides enhanced the superoxide dismutase activity of oysters. Isoproturon enhanced catalase activity, and metconazole enhanced peroxiredoxin activity. Overall, our results show that environmental concentrations of metconazole or isoproturon induced subtle changes in the energy and antioxidant metabolisms of oysters.
Asunto(s)
Antioxidantes/metabolismo , Metabolismo Energético/efectos de los fármacos , Ostreidae/efectos de los fármacos , Plaguicidas/toxicidad , Contaminantes Químicos del Agua/toxicidad , Adenilato Quinasa/metabolismo , Animales , Relación Dosis-Respuesta a Droga , Ostreidae/metabolismoRESUMEN
Since European sea bass (Dicentrarchus labrax) larvae occurred in coastal and estuarine waters at early life stages, they are likely to be exposed to reduced dissolved oxygen waters at a sensitive developmental stage. However, the effects of hypoxia at larval stage, which depend in part on fish species, remain very poorly documented in European sea bass. In the present study, the impacts of an experimental exposure to a chronic moderate hypoxia (40 % air saturation) between 30 and 38 days post-hatching on the physiological and developmental traits of European sea bass larvae were assessed. This study was based on the investigation of survival and growth rates, parameters related to energy metabolism [Citrate Synthase (CS) and Cytochrome-c Oxidase (COX) activities], and biological indicators of the maturation of digestive function [pancreatic (trypsin, amylase) and intestinal (Alkaline Phosphatase "AP" and Aminopeptidase-N "N-LAP") enzymes activities]. While condition of hypoxia exposure did not induce any significant mortality event, lower growth rate as well as CS/COX activity ratio was observed in the Hypoxia Treatment group. In parallel, intestinal enzyme activities were also lower under hypoxia. Altogether, the present data suggest that sea bass larvae cope with moderate hypoxia by (1) reducing processes that are costly in energy and (2) regulating mitochondria functions in order to respond to energy-demand conditions. Both these effects are associated with a delay in the maturation of the digestive function.
Asunto(s)
Lubina/fisiología , Metabolismo Energético/fisiología , Enfermedades de los Peces/fisiopatología , Hipoxia/veterinaria , Mitocondrias/metabolismo , Fosfatasa Alcalina/metabolismo , Aminopeptidasas/metabolismo , Amilasas/metabolismo , Análisis de Varianza , Animales , Complejo IV de Transporte de Electrones/metabolismo , Hipoxia/fisiopatología , Larva/crecimiento & desarrollo , Análisis de Supervivencia , Tripsina/metabolismoRESUMEN
The hermaphrodite Pacific oyster Crassostrea gigas displays a high energy allocation to reproduction. We studied the expression of AMP-activated protein kinase (AMPK) during gametogenesis in the gonad and characterized the mRNA sequences of the AMPK subunits: the AMPK alpha mRNA sequence was previously characterized; we identified AMPK beta, AMPK gamma, and mRNAs of putative AMPK-related targets following bioinformatics mining on existing genomic resources. We analyzed the mRNA expression of the AMPK alpha, beta, and gamma subunits in the gonads of male and female oysters through a reproductive cycle, and we quantified the mRNA expression of genes belonging to fatty acid and glucose metabolism. AMPK alpha mRNA levels were more abundant in males at the first stage of gametogenesis, when mitotic activity and the differentiation of germinal cells occur, and were always more abundant in males than in females. Some targets of fatty acid and glucose metabolism appeared to be correlated with the expression of AMPK subunits at the mRNA level. We then analyzed the sex-specific AMPK activity by measuring the phosphorylation of the catalytic AMPK alpha protein and its expression at the protein level. Both the amount of AMPK alpha protein and threonine 172 phosphorylation appeared to be almost totally inhibited in mature female gonads at stage 3, at the time when accumulation of reserves in oocytes was promoted, while it remained at a high level in mature spermatozoa. Its activation might play a sex-dependent role in the management of energy during gametogenesis in oyster.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Crassostrea/fisiología , Gametogénesis , Regulación del Desarrollo de la Expresión Génica , Gónadas/metabolismo , Subunidades de Proteína/metabolismo , Proteínas Quinasas Activadas por AMP/biosíntesis , Proteínas Quinasas Activadas por AMP/genética , Animales , Acuicultura , Biología Computacional , Minería de Datos , Metabolismo Energético , Activación Enzimática , Femenino , Francia , Gónadas/citología , Gónadas/crecimiento & desarrollo , Masculino , Fosforilación , Filogenia , Procesamiento Proteico-Postraduccional , Subunidades de Proteína/biosíntesis , Subunidades de Proteína/genética , ARN Mensajero/metabolismo , Caracteres Sexuales , Treonina/metabolismoRESUMEN
In intertidal zones, species such as sessile shellfish exhibit extended phenotypic plasticity to face rapid environmental changes, but whether frequent exposure to intertidal limits of the distribution range impose physiological costs for the animal remains elusive. Here, we explored how phenotypic plasticity varied along foreshore range at multiple organization levels, from molecular to cellular and whole organism acclimatization, in the Pacific oyster (Crassostrea gigas). We exposed 7-month-old individuals for up to 16 months to three foreshore levels covering the vertical range for this species, representing 20, 50 and 80% of the time spent submerged monthly. Individuals at the upper range limit produced energy more efficiently, as seen by steeper metabolic reactive norms and unaltered ATP levels despite reduced mitochondrial density. By spending most of their time emerged, oysters mounted an antioxidant shielding concomitant with lower levels of pro-oxidant proteins and postponed age-related telomere attrition. Instead, individuals exposed at the lower limit range near subtidal conditions showed lower energy efficiencies, greater oxidative stress and shorter telomere length. These results unraveled the extended acclimatization strategies and the physiological costs of living too fast in subtidal conditions for an intertidal species.
RESUMEN
The Pacific oyster Crassostrea gigas is established in the marine intertidal zone, experiencing rapid and highly dynamic environmental changes throughout the tidal cycle. Depending on the bathymetry, oysters face oxygen deprivation, lack of nutrients, and high changes in temperature during alternation of the cycles of emersion/immersion. Here we showed that intertidal oysters at a bathymetry level of 3 and 5 m delayed by ten days the onset of mortality associated with Pacific Oyster Mortality Syndrome (POMS) as compared to subtidal oysters. Intertidal oysters presented a lower growth but similar energetic reserves to subtidal oysters but induced proteomic changes indicative of a boost in metabolism, inflammation, and innate immunity that may have improved their resistance during infection with the Ostreid herpes virus. Our work highlights that intertidal harsh environmental conditions modify host-pathogen interaction and improve oyster health. This study opens new perspectives on oyster farming for mitigation strategies based on tidal height.
Asunto(s)
Crassostrea , Herpesviridae , Animales , Interacciones Huésped-Patógeno , Inmunidad Innata , ProteómicaRESUMEN
Digestive microbiota provide a wide range of beneficial effects on host physiology and are therefore likely to play a key role in marine intertidal bivalve ability to acclimatize to the intertidal zone. This study investigated the effect of intertidal levels on the digestive bacterial microbiota of oysters (Crassostrea gigas) and clams (Ruditapes philippinarum), two bivalves with different ecological niches. Based on 16S rRNA region sequencing, digestive glands, seawater and sediments harbored specific bacterial communities, dominated by operational taxonomic units assigned to the Mycoplasmatales,Desulfobacterales and Rhodobacterales orders, respectively. Field implantation modified digestive bacterial microbiota of both bivalve species according to their intertidal position. Rhodospirillales and Legionellales abundances increased in oysters and clams from the low intertidal level, respectively. After a 14-day depuration process, these effects were still observed, especially for clams, while digestive bacterial microbiota of oysters were subjected to more short-term environmental changes. Nevertheless, 3.5 months stay on an intertidal zone was enough to leave an environmental footprint on the digestive bacterial microbiota, suggesting the existence of autochthonous bivalve bacteria. When comparing clams from the three intertidal levels, 20% of the bacterial assemblage was shared among the levels and it was dominated by an operational taxonomic unit affiliated to the Mycoplasmataceae and Spirochaetaceae families.
Asunto(s)
Bivalvos , Crassostrea , Microbiota , Animales , Bacterias/genética , Humanos , ARN Ribosómico 16S/genética , Agua de MarRESUMEN
BACKGROUND: Although bivalves are among the most-studied marine organisms because of their ecological role and economic importance, very little information is available on the genome sequences of oyster species. This report documents three large-scale cDNA sequencing projects for the Pacific oyster Crassostrea gigas initiated to provide a large number of expressed sequence tags that were subsequently compiled in a publicly accessible database. This resource allowed for the identification of a large number of transcripts and provides valuable information for ongoing investigations of tissue-specific and stimulus-dependant gene expression patterns. These data are crucial for constructing comprehensive DNA microarrays, identifying single nucleotide polymorphisms and microsatellites in coding regions, and for identifying genes when the entire genome sequence of C. gigas becomes available. DESCRIPTION: In the present paper, we report the production of 40,845 high-quality ESTs that identify 29,745 unique transcribed sequences consisting of 7,940 contigs and 21,805 singletons. All of these new sequences, together with existing public sequence data, have been compiled into a publicly-available Website http://public-contigbrowser.sigenae.org:9090/Crassostrea_gigas/index.html. Approximately 43% of the unique ESTs had significant matches against the SwissProt database and 27% were annotated using Gene Ontology terms. In addition, we identified a total of 208 in silico microsatellites from the ESTs, with 173 having sufficient flanking sequence for primer design. We also identified a total of 7,530 putative in silico, single-nucleotide polymorphisms using existing and newly-generated EST resources for the Pacific oyster. CONCLUSION: A publicly-available database has been populated with 29,745 unique sequences for the Pacific oyster Crassostrea gigas. The database provides many tools to search cleaned and assembled ESTs. The user may input and submit several filters, such as protein or nucleotide hits, to select and download relevant elements. This database constitutes one of the most developed genomic resources accessible among Lophotrochozoans, an orphan clade of bilateral animals. These data will accelerate the development of both genomics and genetics in a commercially-important species with the highest annual, commercial production of any aquatic organism.
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Crassostrea/genética , Bases de Datos Genéticas , Etiquetas de Secuencia Expresada , Animales , Perfilación de la Expresión Génica , Biblioteca de Genes , Genoma , Genómica/métodos , Repeticiones de Microsatélite , Polimorfismo de Nucleótido Simple , Análisis de Secuencia de ADN , Interfaz Usuario-ComputadorRESUMEN
The purpose of this study was to determine whether n-3 PUFA result in an effect on endothelial function that is in addition to that of acute exercise. For 4 weeks, male Sprague-Dawley rats were subjected to a diet based on n-3 PUFA or a standard diet. In each diet group, ten rats were submitted to an acute treadmill exercise while the remaining ten acted as sedentary controls. The running speed was progressively increased until the animals were exhausted. Endothelial function was then assessed by measuring isometric tension in rings of the thoracic aorta. In vessels precontracted with 0.1 microm-phenylephrine, responses to acetylcholine (ACh) were significantly improved following acute exercise in all diet groups. When PUFA supplementation was compared to the standard diet no significant difference was found in response to ACh, either at rest or after an acute exercise. Pretreatment of rings with Nomega-nitro-l-arginine methyl esther (50 microm) inhibited the ACh-mediated vasorelaxation in all groups. Response to 10 microm-nifedipine, an L-type Ca2+ channel antagonist, was similarly enhanced after acute exercise in both standard and PUFA diets. Furthermore, response to 0.01 microm-nifedipine was significantly higher after acute exercise only in the PUFA diet. In conclusion, in our 'healthy' rat model with 'normal' baseline endothelial function, acute exercise improves response to ACh while PUFA supplementation alone or in combination with acute exercise has no effect on endothelium-dependent vasorelaxation. However, PUFA may potentiate the acute exercise effect on smooth muscle cell relaxation via L-type Ca2+ channel modifications.
Asunto(s)
Endotelio Vascular/fisiología , Ácidos Grasos Omega-3/farmacología , Músculo Liso Vascular/fisiología , Condicionamiento Físico Animal , Vasodilatación/fisiología , Acetilcolina/farmacología , Animales , Aorta , Relación Dosis-Respuesta a Droga , Endotelio Vascular/efectos de los fármacos , Masculino , Músculo Liso Vascular/efectos de los fármacos , Nifedipino/farmacología , Ratas , Ratas Sprague-Dawley , Vasodilatadores/farmacologíaRESUMEN
The Warburg effect is one of the hallmarks of cancer cells in humans. It is a true metabolic reprogramming to aerobic glycolysis, allowing cancer cells to meet their particular energy needs for growth, proliferation, and resistance to apoptosis, depending on the microenvironment they encounter within the tumor. We have recently discovered that the Crassostrea gigas oyster can naturally reprogram its metabolism to the Warburg effect. Thus, the oyster becomes a new invertebrate model useful for cancer research. Due to its lifestyle, the oyster C. gigas has special abilities to adapt its metabolism to the extreme changes in the environment in which it is located. The oyster C. gigas is therefore a model of interest to study how the environment can control the Warburg effect under conditions that could not be explored in vertebrate model species.
Asunto(s)
Crassostrea/fisiología , Modelos Animales de Enfermedad , Neoplasias , Animales , Apoptosis , Proliferación Celular , Microambiente Celular , Reprogramación Celular , GlucólisisRESUMEN
The Pacific oyster, Crassostrea gigas, is an osmoconforming bivalve exposed to wide salinity fluctuations. The physiological mechanisms used by oysters to cope with salinity stress are energy demanding and may impair other processes, such as defense against pathogens. This oyster species has been experiencing recurrent mortality events caused by the Ostreid herpesvirus 1 (OsHV-1). The objectives of this study were to investigate the effect of salinity (10, 15, 25 and 35) on energetic reserves, key enzyme activities and membrane fatty acids, and to identify the metabolic risk factors related to OsHV-1-induced mortality of oysters. Acclimation to low salinity led to increased water content, protein level, and energetic reserves (carbohydrates and triglycerides) of oysters. The latter was consistent with lower activity of hexokinase, the first enzyme involved in glycolysis, up-regulation of AMP-activated protein kinase, a major regulator of cellular energy metabolism, and lower activity of catalase, an antioxidant enzyme involved in management of reactive oxygen species. Acclimation to salinity also involved a major remodeling of membrane fatty acids. Particularly, 20:4n-6 decreased linearly with decreasing salinity, likely reflecting its mobilization for prostaglandin synthesis in oysters. The survival of oysters exposed to OsHV-1 varied from 43% to 96% according to salinity ( Fuhrmann et al., 2016). Risk analyses showed that activity of superoxide dismutase and levels of proteins, carbohydrates, and triglycerides were associated with a reduced risk of death. Therefore, animals with a higher antioxidant activity and a better physiological condition seemed less susceptible to OsHV-1.
RESUMEN
Voltage-dependent anion channel (VDAC) is a key mitochondrial protein. VDAC drives cellular energy metabolism by controlling the influx and efflux of metabolites and ions through the mitochondrial membrane, playing a role in its permeabilization. This protein exerts a pivotal role during the white spot syndrome virus (WSSV) infection in shrimp, through its involvement in a particular metabolism that plays in favor of the virus, the Warburg effect. The Warburg effect corresponds to an atypical metabolic shift toward an aerobic glycolysis that provides energy for rapid cell division and resistance to apoptosis. In the Pacific oyster Crassostrea gigas, the Warburg effect occurs during infection by Ostreid herpesvirus (OsHV-1). At present, the role of VDAC in the Warburg effect, OsHV-1 infection and apoptosis is unknown. Here, we developed a specific antibody directed against C. gigas VDAC. This tool allowed us to quantify the tissue-specific expression of VDAC, to detect VDAC oligomers, and to follow the amount of VDAC in oysters deployed in the field. We showed that oysters sensitive to a mortality event in the field presented an accumulation of VDAC. Finally, we propose to use VDAC quantification as a tool to measure the oyster susceptibility to OsHV-1 depending on its environment.
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Crassostrea/virología , Infecciones por Herpesviridae/metabolismo , Herpesviridae/metabolismo , Canales Aniónicos Dependientes del Voltaje/química , Animales , Anticuerpos , Apoptosis , Crassostrea/metabolismo , Infecciones por Herpesviridae/fisiopatología , Análisis de Secuencia de Proteína , Canales Aniónicos Dependientes del Voltaje/inmunologíaRESUMEN
The king scallop Pecten maximus is a high valuable species of great interest in Europe for both fishery and aquaculture. Notably, there has been an increased investment to produce seed for enhancement programmes of wild scallop populations. However, hatchery production is a relatively new industry and it is still underdeveloped. Major hurdles are spawning control and gamete quality. In the present study, a total of 14 scallops were sampled in the bay of Brest (Brittany, France) to compare transcriptomic profiles of mature oocytes collected by spawning induction or by stripping. To reach such a goal, a microarray analysis was performed by using a custom 8x60K oligonucleotide microarray representing 45,488 unique scallop contigs. First we identified genes that were differentially expressed depending on oocyte quality, estimated as the potential to produce D-larvae. Secondly, we investigated the transcriptional features of both stripped and spawned oocytes. Genes coding for proteins involved in cytoskeletal dynamics, serine/threonine kinases signalling pathway, mRNA processing, response to DNA damage, apoptosis and cell-cycle appeared to be of crucial importance for both oocyte maturation and developmental competence. This study allowed us to dramatically increase the knowledge about transcriptional features of oocyte quality and maturation, as well as to propose for the first time putative molecular markers to solve a major bottleneck in scallop aquaculture.
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Perfilación de la Expresión Génica , Oocitos/crecimiento & desarrollo , Oocitos/metabolismo , Pecten/crecimiento & desarrollo , Pecten/genética , Animales , Femenino , Fertilidad , Larva/genética , Masculino , Oocitos/fisiología , Transcripción GenéticaRESUMEN
Oysters play an important role in estuarine and coastal marine habitats, where the majority of humans live. In these ecosystems, environmental degradation is substantial, and oysters must cope with highly dynamic and stressful environmental constraints during their lives in the intertidal zone. The availability of the genome sequence of the Pacific oyster Crassostrea gigas represents a unique opportunity for a comprehensive assessment of the signal transduction pathways that the species has developed to deal with this unique habitat. We performed an in silico analysis to identify, annotate and classify protein kinases in C. gigas, according to their kinase domain taxonomy classification, and compared with kinome already described in other animal species. The C. gigas kinome consists of 371 protein kinases, making it closely related to the sea urchin kinome, which has 353 protein kinases. The absence of gene redundancy in some groups of the C. gigas kinome may simplify functional studies of protein kinases. Through data mining of transcriptomes in C. gigas, we identified part of the kinome which may be central during development and may play a role in response to various environmental factors. Overall, this work contributes to a better understanding of key sensing pathways that may be central for adaptation to a highly dynamic marine environment.
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Crassostrea/enzimología , Crassostrea/genética , Ambiente , Regulación del Desarrollo de la Expresión Génica , Genómica , Proteínas Quinasas/genética , Animales , Crassostrea/crecimiento & desarrollo , Activación Enzimática , Filogenia , Proteínas Quinasas/metabolismoRESUMEN
We investigated the effect of brown ring disease (BRD) development and algal diet on energy reserves and activity of enzymes related to energy metabolism, antioxidant system and immunity in Manila clam, Ruditapes philippinarum. We found that algal diet did not impact the metabolic response of clams exposed to Vibrio tapetis. At two days post-injection (dpi), activities of superoxide dismutase and glutathione peroxidase (GPx) decreased whereas activities of nitric oxide synthase (iNOS) and catalase increased in infected clams, although no clinical signs were visible (BRD-). At 7 dpi, activities of several antioxidant and immune-related enzymes were markedly increased in BRD-likely indicating an efficient reactive oxygen species (ROS) scavenging compared to animals which developed clinical signs of BRD (BRD+). Therefore, resistance to BRD clinical signs appearance was associated with higher detoxification of ROS and enhancement of immune response. This study provides new biochemical indicators of disease resistance and a more comprehensive view of the global antioxidant response of clam to BRD development.