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1.
Sci Total Environ ; 806(Pt 1): 149925, 2022 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-34555605

RESUMEN

Expected effects on marine biota of the ongoing elevation of water temperature and high latitudes is of major concern when considering the reliability of coastal ecosystem production. To compare the capacity of coastal organisms to cope with a temperature increase depending on their environmental history, responses of adult blue mussels (Mytilus spp.) taken from two sites differentially exposed to chemical pollution were investigated during an experimental exposure to a thermal stress. Immune parameters were notably altered by extreme warming and transcriptional changes for a broad selection of genes were associated to the temperature increase following a two-step response pattern. Site-specific responses suggested an influence of environmental history and support the possibility of a genetic basis in the physiological response. However no meaningful difference was detected between the response of hybrids and M galloprovincialis. This study brings new information about the capacity of mussels to cope with the ongoing elevation of water temperature in these coastal ecosystems.


Asunto(s)
Mytilus edulis , Mytilus , Contaminantes Químicos del Agua , Animales , Ecosistema , Reproducibilidad de los Resultados , Estrés Fisiológico , Contaminantes Químicos del Agua/toxicidad
2.
Mar Genomics ; 50: 100704, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-31506217

RESUMEN

In recent years, numerous studies conducted on teleost fish have highlighted the contribution of transcriptomic studies in elucidating the physiological mechanisms underlying the molecular events of oogenesis and follicular atresia, enabling the identification of potential genes and molecular networks that participate in both the reproductive cycle and the process of follicular atresia. Atresia can affect the reproductive potential of females by reducing the healthy eggs that a female can spawn in both aquaculture and wild populations. The substantial diversity of reproductive strategies exhibited by teleost fish has contributed to the difficulty in identifying common genes between species, but a set of core genes has emerged as potential markers for atresia in relation to apoptosis/autophagy, lipid metabolism, oxidative metabolism and other physiological processes similar to those identified in other vertebrates, even mammals. We review the current status of the genes that have been identified in ovaries with atretic oocytes. Our primary goal is to review the current status regarding gene expression during gonadal development and follicular atresia. This information will enable us to understand the factors and expression patterns involved in the follicular atresia of teleost fish.


Asunto(s)
Peces/fisiología , Atresia Folicular/genética , Expresión Génica , Ovario/metabolismo , Animales , Femenino , Peces/genética , Ovario/crecimiento & desarrollo
3.
Artículo en Inglés | MEDLINE | ID: mdl-26282335

RESUMEN

It is commonly known that the nature of the diet has diverse consequences on larval performance and longevity, however it is still unclear which genes have critical impacts on bivalve development and which pathways are of particular importance in their vulnerability or resistance. First we show that a diet deficient in essential fatty acid (EFA) produces higher larval mortality rates, a reduced shell growth, and lower postlarval performance, all of which are positively correlated with a decline in arachidonic and eicosapentaenoic acids levels, two EFAs known as eicosanoid precursors. Eicosanoids affect the cell inflammatory reactions and are synthesized from long-chain EFAs. Second, we show for the first time that a deficiency in eicosanoid precursors is associated with a network of 29 genes. Their differential regulation can lead to slower growth and higher mortality of Mytilus edulis larvae. Some of these genes are specific to bivalves and others are implicated at the same time in lipid metabolism and defense. Several genes are expressed only during pre-metamorphosis where they are essential for muscle or neurone development and biomineralization, but only in stress-induced larvae. Finally, we discuss how our networks of differentially expressed genes might dynamically alter the development of marine bivalves, especially under dietary influence.


Asunto(s)
Bivalvos/crecimiento & desarrollo , Bivalvos/genética , Ácidos Grasos Esenciales/metabolismo , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Bivalvos/fisiología , Dieta , Eicosanoides/metabolismo , Regulación del Desarrollo de la Expresión Génica , Redes Reguladoras de Genes , Larva/genética , Larva/crecimiento & desarrollo , Larva/fisiología , Aprendizaje Automático , Modelos Biológicos
4.
Aquat Toxicol ; 162: 126-137, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25814057

RESUMEN

Brest harbor (Bay of Brest, Brittany, France) has a severe past of anthropogenic chemical contamination, but inputs tended to decrease, indicating a reassessment of its ecotoxicological status should be carried out. Here, native and caged mussels (Mytilus spp.) were used in combination to evaluate biological effects of chronic chemical contamination in Brest harbor. Polycyclic aromatic hydrocarbon (PAH) contamination was measured in mussel tissues as a proxy of harbor and urban pollution. Biochemical biomarkers of xenobiotic biotransformation, antioxidant defenses, generation of reducing equivalents, energy metabolism and oxidative damage were studied in both gills and digestive glands of native and caged mussels. In particular, activities of glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), NADP-dependent isocitrate dehydrogenase (IDP), pyruvate kinase (PK) and phosphoenolpyruvate carboxykinase (PEPCK) were measured and lipid peroxidation was assessed by malondialdehyde (MDA) quantification. In addition, a condition index was calculated to assess the overall health of the mussels. Moderate PAH contamination was detected in digestive glands of both native and caged individuals from the exposed site. Modulations of biomarkers were detected in digestive glands of native harbor mussels indicating the presence of a chemical pressure. In particular, results suggested increased biotransformation (GST), antioxidant defenses (CAT), NADPH generation (IDP) and gluconeogenesis (PEPCK), which could represent a coordinated response against chemically-induced cellular stress. Lipid peroxidation assessment and condition index indicated an absence of acute stress in the same mussels suggesting metabolic changes could, at least partially, offset the negative effects of contamination. In caged mussels, only GR was found modulated compared to non-exposed mussels but significant differences in oxidative stress and energy-related biomarkers were observed compared to native harbor mussels. Overall, these results suggested mussels chronically exposed to contamination have set up metabolic adaptation, which may contribute to their survival in the moderately contaminated harbor of Brest. Whether these adaptive traits result from phenotypic plasticity or genetic adaptation needs to be further investigated.


Asunto(s)
Adaptación Fisiológica/efectos de los fármacos , Monitoreo del Ambiente/métodos , Mytilus/efectos de los fármacos , Hidrocarburos Policíclicos Aromáticos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Contaminación Química del Agua/efectos adversos , Animales , Biomarcadores/metabolismo , Biotransformación , Metabolismo Energético/efectos de los fármacos , Francia , Peroxidación de Lípido/efectos de los fármacos , Mytilus/metabolismo , Estrés Oxidativo/efectos de los fármacos , Hidrocarburos Policíclicos Aromáticos/metabolismo , Contaminantes Químicos del Agua/metabolismo
5.
Dev Comp Immunol ; 51(2): 287-97, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25766281

RESUMEN

Vibrio harveyi is a marine bacterial pathogen responsible for episodic abalone mortalities in France, Japan and Australia. In the European abalone, V. harveyi invades the circulatory system in a few hours after exposure and is lethal after 2 days of infection. In this study, we investigated the responses of European abalone immune cells over the first 24 h of infection. Results revealed an initial induction of immune gene expression including Rel/NF-kB, Mpeg and Clathrin. It is rapidly followed by a significant immuno-suppression characterized by reduced cellular hemocyte parameters, immune response gene expressions and enzymatic activities. Interestingly, Ferritin was overexpressed after 24 h of infection suggesting that abalone attempt to counter V. harveyi infection using soluble effectors. Immune function alteration was positively correlated with V. harveyi concentration. This study provides the evidence that V. harveyi has a hemolytic activity and an immuno-suppressive effect in the European abalone.


Asunto(s)
Ferritinas/metabolismo , Gastrópodos/inmunología , Hemocitos/inmunología , Vibriosis/inmunología , Vibrio/inmunología , Animales , Clatrina/genética , Clatrina/metabolismo , Europa (Continente) , Ferritinas/genética , Regulación de la Expresión Génica , Hemocitos/microbiología , Hemólisis , Inmunidad , Inmunomodulación , FN-kappa B/genética , FN-kappa B/metabolismo , Proteínas Oncogénicas v-rel/genética , Proteínas Oncogénicas v-rel/metabolismo
6.
Appl Environ Microbiol ; 80(20): 6328-33, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25107972

RESUMEN

Vibrio harveyi is a marine bacterial pathogen responsible for episodic epidemics generally associated with massive mortalities in many marine organisms, including the European abalone Haliotis tuberculata. The aim of this study was to identify the portal of entry and the dynamics of infection of V. harveyi in the European abalone. The results indicate that the duration of contact between V. harveyi and the European abalone influences the mortality rate and precocity. Immediately after contact, the epithelial and mucosal area situated between the gills and the hypobranchial gland was colonized by V. harveyi. Real-time PCR analyses and culture quantification of a green fluorescent protein-tagged strain of V. harveyi in abalone tissues revealed a high density of bacteria adhering to and then penetrating the whole gill-hypobranchial gland tissue after 1 h of contact. V. harveyi was also detected in the hemolymph of a significant number of European abalones after 3 h of contact. In conclusion, this article shows that a TaqMan real-time PCR assay is a powerful and useful technique for the detection of a marine pathogen such as V. harveyi in mollusk tissue and for the study of its infection dynamics. Thus, we have revealed that the adhesion and then the penetration of V. harveyi in European abalone organs begin in the first hours of contact. We also hypothesize that the portal of entry of V. harveyi in the European abalone is the area situated between the gills and the hypobranchial gland.


Asunto(s)
Gastrópodos/microbiología , Interacciones Huésped-Patógeno , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Vibrio/patogenicidad , Animales , Branquias/microbiología , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Hemolinfa/microbiología , Sensibilidad y Especificidad , Factores de Tiempo , Vibrio/genética , Vibriosis/microbiología , Vibriosis/mortalidad , Vibriosis/veterinaria
7.
Gene ; 551(1): 65-78, 2014 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-25158132

RESUMEN

Understanding the mechanisms that enable growth and survival of an organism while driving it to the full range of its adaptation is fundamental to the issues of biodiversity and evolution, particularly regarding global climatic changes. Here we report the Illumina RNA-sequencing (RNA-seq) and de novo assembly of the blue mussel Mytilus edulis transcriptome during early development. This study is based on high-throughput data, which associates genome-wide differentially expressed transcript (DET) patterns with early activation of developmental processes. Approximately 50,383 high-quality contigs were assembled. Over 8000 transcripts were associated with functional proteins from public databases. Coding and non-coding genes served to design customized microarrays targeting every developmental stage, which encompass major transitions in tissue organization. Consequently, multi-processing pattern exploration protocols applied to 3633 DETs helped discover 12 unique coordinated eigengenes supposedly implicated in various physiological and morphological changes that larvae undergo during early development. Moreover, dynamic Bayesian networks (DBNs) provided key insights to understand stage-specific molecular mechanisms activated throughout ontogeny. In addition, delayed and contemporaneous interactions between DETs were coerced with 16 relevant regulators that interrelated in non-random genetic regulatory networks (GRNs). Genes associated with mechanisms of neural and muscular development have been characterized and further included in dynamic networks necessary in growth and functional morphology. This is the first large-scale study being dedicated to M. edulis throughout early ontogeny. Integration between RNA-seq and microarray data enabled a high-throughput exploration of hidden processes essential in growth and survival of microscopic mussel larvae. Our integrative approach will support a holistic understanding of systems biology and will help establish new links between environmental assessment and functional development of marine bivalves.


Asunto(s)
Redes Reguladoras de Genes , Mytilus edulis/crecimiento & desarrollo , Mytilus edulis/genética , Animales , Teorema de Bayes , Ensamble y Desensamble de Cromatina , Lógica Difusa , Regulación del Desarrollo de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Larva/genética , Metamorfosis Biológica/genética , Análisis de Componente Principal , Transcriptoma
8.
Fish Shellfish Immunol ; 36(1): 1-8, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24215911

RESUMEN

Wild or farmed abalone are regularly exposed to stressors, such as air exposure and handling. Immune and transcriptional responses as well as susceptibility to vibriosis of sexually mature or immature European abalone acclimated at 16 or 19 °C were determined following handling or air exposure. Hemocyte density and H2O2 production increased while hemocyte viability and phagocytic index decreased following handling. Air exposure induces a decrease of hemocyte density and phagocytic index. Measurement of the expression of genes implicated in general metabolic, immunological and stress responses in gills, foot-muscle and hemocytes by real time q-PCR suggested that both stressors lead to a metabolic rate depression, characterized by a general inhibition of transcription. Finally, following handling a Vibrio harveyi challenge enhances almost 100% mortality of sexually immature animals at 19 °C while it has been previously demonstrated that only mature are susceptible to vibriosis.


Asunto(s)
Gastrópodos/inmunología , Hemocitos/inmunología , Estrés Fisiológico/inmunología , Transcripción Genética/inmunología , Vibriosis/inmunología , Vibrio/inmunología , Animales , Femenino , Gastrópodos/genética , Gastrópodos/virología , Perfilación de la Expresión Génica/métodos , Hemocitos/virología , Estimación de Kaplan-Meier , Masculino , ARN/química , ARN/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Estrés Fisiológico/genética , Vibriosis/virología
9.
PLoS One ; 8(5): e64534, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23704993

RESUMEN

BACKGROUND: Bacterial infections are common in bivalve larvae and can lead to significant mortality, notably in hatcheries. Numerous studies have identified the pathogenic bacteria involved in such mortalities, but physiological changes associated with pathogen exposure at larval stage are still poorly understood. In the present study, we used an integrative approach including physiological, enzymatic, biochemical, and molecular analyses to investigate changes in energy metabolism, lipid remodelling, cellular stress, and immune status of Crassostrea gigas larvae subjected to experimental infection with the pathogenic bacteria Vibrio coralliilyticus. FINDINGS: Our results showed that V. coralliilyticus exposure induced (1) limited but significant increase of larvae mortality compared with controls, (2) declined feeding activity, which resulted in energy status changes (i.e. reserve consumption, ß-oxidation, decline of metabolic rate), (3) fatty acid remodeling of polar lipids (changes in phosphatidylinositol and lysophosphatidylcholine composition`, non-methylene-interrupted fatty acids accumulation, lower content of major C20 polyunsaturated fatty acids as well as activation of desaturases, phospholipase and lipoxygenase), (4) activation of antioxidant defenses (catalase, superoxide dismutase, peroxiredoxin) and cytoprotective processes (heat shock protein 70, pernin), and (5) activation of the immune response (non-self recognition, NF-κκ signaling pathway, haematopoiesis, eiconosoids and lysophosphatidyl acid synthesis, inhibitor of metalloproteinase and antimicrobial peptides). CONCLUSION: Overall, our results allowed us to propose an integrative view of changes induced by a bacterial infection in Pacific oyster larvae, opening new perspectives on the response of marine bivalve larvae to infections.


Asunto(s)
Infecciones Bacterianas/microbiología , Infecciones Bacterianas/fisiopatología , Crassostrea/microbiología , Crassostrea/fisiología , Vibrio/fisiología , Animales , Crassostrea/genética , Electroforesis en Gel de Gradiente Desnaturalizante , Metabolismo Energético , Larva/microbiología , Larva/fisiología , Modelos Biológicos , Estrés Oxidativo , Océano Pacífico , Análisis de Supervivencia
10.
J Invertebr Pathol ; 113(2): 163-76, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23535139

RESUMEN

The abalone production in Chile has increased considerably in recent years with no sign of tapering off. Open and semi-closed circuits in the marine water zones in the north and south of Chile are the preferred areas of culture. Coastal ecosystems are subjected to a wide variety of contaminants that generate stress that affects populations via their impacts to individuals at both physiological and genetic levels. This work investigated the genomic and cellular response of post-weaning juvenile Haliotis rufescens abalone under hatchery conditions, fed with probiotic diets, and subsequently challenged with Vibrio parahaemolyticus. The expression patterns of 16 selected genes associated with different metabolic pathways were analyzed using Real-Time PCR. Gene expression was then compared to immunological response parameters in the abalone and quantification of V. parahaemolyticus during the experimental period. Both transcriptomic and immunological analyses indicated significant alteration of physiological processes in H. rufescens correlated to exposure to the pathogenic bacteria, as well as to probiotic nutrition.


Asunto(s)
Gastrópodos/microbiología , Interacciones Huésped-Patógeno/genética , Fagocitosis/efectos de los fármacos , Probióticos/farmacología , Vibrio parahaemolyticus/fisiología , Animales , Dieta , Gastrópodos/efectos de los fármacos , Gastrópodos/fisiología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Transcriptoma/efectos de los fármacos
11.
J Exp Biol ; 216(Pt 9): 1561-9, 2013 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-23307802

RESUMEN

As oxygen concentrations in marine coastal habitats can fluctuate rapidly and drastically, sessile marine organisms such as the oyster Crassostrea gigas can experience marked and rapid oxygen variations. In this study, we investigated the responses of oyster gill mitochondria to short-term hypoxia (3 and 12 h, at 1.7 mg O2 l(-1)) and subsequent re-oxygenation. Mitochondrial respiratory rates (states 3 and 4 stimulated by glutamate) and phosphorylation efficiency [respiratory control ratio (RCR) and the relationship between ADP and oxygen consumption (ADP/O)] were measured. Cytochrome c oxidase (CCO) activity and cytochrome concentrations (a, b, c1 and c) were measured to investigate the rearrangements of respiratory chain subunits. The potential implication of an alternative oxidase (AOX) was investigated using an inhibitor of the respiratory chain (antimycin A) and through gene expression analysis in gills and digestive gland. Results indicate a downregulation of mitochondrial capacity, with 60% inhibition of respiratory rates after 12 h of hypoxia. RCR remained stable, while ADP/O increased after 12 h of hypoxia and 1 h of re-oxygenation, suggesting increased phosphorylation efficiency. CCO showed a fast and remarkable increase of its catalytic activity only after 3 h of hypoxia. AOX mRNA levels showed similar patterns in gills and digestive gland, and were upregulated after 12 and 24 h of hypoxia and during re-oxygenation. Results suggest a set of controls regulating mitochondrial functions in response to oxygen fluctuations, and demonstrate the fast and extreme plasticity of oyster mitochondria in response to oxygen variations.


Asunto(s)
Crassostrea/metabolismo , Mitocondrias/metabolismo , Oxígeno/farmacología , Anaerobiosis/efectos de los fármacos , Animales , Respiración de la Célula/efectos de los fármacos , Crassostrea/efectos de los fármacos , Crassostrea/enzimología , Citocromos/metabolismo , Sistema Digestivo/enzimología , Complejo IV de Transporte de Electrones/metabolismo , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Branquias/enzimología , Mitocondrias/efectos de los fármacos , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Consumo de Oxígeno/efectos de los fármacos , Océano Pacífico , Fosforilación/efectos de los fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factores de Tiempo
12.
Gene ; 499(1): 70-5, 2012 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-22417898

RESUMEN

Quantification of mRNA of genes related to metabolism, immunity and cellular stress was examined in relation to a massive mortality event during the culture of American oyster larvae, Crassostrea virginica which was probably, in regard to previous microbiological analysis, induced by Vibrio infection. To document molecular changes associated with the mortality event, mRNA levels were compared to biochemical and physiological data, previously described in a companion paper. Among the 18 genes studied, comparatively to the antibiotic control, 10 showed a lower relative gene expression when the massive mortality occurred. Six of them are presumed to be related to metabolism, corroborating the metabolic depression associated with the mortality event suggested by biochemical and physiological analyses. Relationships between the regulation of antioxidant enzyme activities, lipid peroxidation, and the mRNA abundance of genes linked to oxidative stress, cytoprotection, and immune response are also discussed. Finally, we observed an increase in the transcript abundance of two genes involved in apoptosis and cell regulation simultaneously with mortality, suggesting that these processes might be linked.


Asunto(s)
Crassostrea/genética , Crassostrea/inmunología , Crassostrea/metabolismo , Fenómenos del Sistema Inmunológico/genética , Estrés Fisiológico/genética , Animales , Infecciones Bacterianas/genética , Infecciones Bacterianas/inmunología , Infecciones Bacterianas/mortalidad , Secuencia de Bases , Biomarcadores/análisis , Biomarcadores/metabolismo , Crassostrea/fisiología , Metabolismo Energético/genética , Metabolismo Energético/inmunología , Metabolismo Energético/fisiología , Regulación del Desarrollo de la Expresión Génica , Estudios de Asociación Genética , Larva/genética , Larva/inmunología , Larva/metabolismo , Larva/fisiología , Metabolismo/genética , Mortalidad
13.
Biomarkers ; 17(1): 85-95, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-22149898

RESUMEN

Summer mortality events have been observed in Pacific oyster Crassostrea gigas for several decades. This paper examines the selective pressure exerted by summer mortality on the polymorphism of a newly identified oyster metallothionein gene. CgMT4 cDNA and genomic sequences were obtained. CgMT4 was studied in two generations of oysters reared in three sites on the French Atlantic coast, using single strand conformation polymorphism analysis. Four alleles were detected. Individuals carrying genotype MT4-CD seem to have higher susceptibility to summer risk conditions. The MT4 gene could be a potential new genetic marker for susceptibility; further validation studies are recommended.


Asunto(s)
Crassostrea/fisiología , Metalotioneína/genética , Polimorfismo Genético , Selección Genética , Estrés Fisiológico/genética , Alelos , Animales , Secuencia de Bases , Crassostrea/clasificación , Crassostrea/genética , Marcadores Genéticos , Genotipo , Datos de Secuencia Molecular , Filogenia , Estaciones del Año
14.
Aquat Toxicol ; 109: 70-9, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22204990

RESUMEN

Genetic and ecophysiological responses of oyster, Crassostrea gigas, populations to environmental stress were investigated in three highly contaminated French estuaries (the Gironde, Loire and Vilaine) and compared to a control, the Belon estuary. A strong response in both metallothionein CgMT4 mRNA expression, as determined by semi-quantitative RT-PCR, and amount of protein, as determined by ELISA tests, was demonstrated in estuarine populations subjected to differential contamination, with an inhibition in the area most highly contaminated with metals. In these same estuarine populations, we found polymorphisms of the metallothionein CgMT4 gene and three other genes (glutamine synthetase--GS, delta-9 desaturase--D9 and phosphoglucomutase--PGM) involved in stress response of C. gigas. We showed that genetic differentiation was observed for MT4 and PGM genes in the Gironde estuary which is highly contaminated with metals. A strong seasonal effect was observed. Phenotype-genotype coupling revealed that one particular MT4 allele and one PGM allele seemed to be associated with metal sensitivity expressed as lower detoxification efficiency and higher metal bioaccumulation. The MT4 gene is a good physiological and genetic marker of stress response and susceptibility.


Asunto(s)
Crassostrea/efectos de los fármacos , Crassostrea/fisiología , Ambiente , Estrés Fisiológico , Contaminantes Químicos del Agua/toxicidad , Alelos , Animales , Francia , Regulación de la Expresión Génica/efectos de los fármacos , Variación Genética , Genotipo , Metalotioneína/genética , Metalotioneína/metabolismo , Metales Pesados/toxicidad , Estaciones del Año , Estrés Fisiológico/genética
15.
Mar Pollut Bull ; 62(12): 2671-80, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22001298

RESUMEN

This study reports molecular markers potentially associated with resistance or sensitivity to the impact of copper in juvenile red abalone, Haliotis rufescens, in the north of Chile under experimental conditions. Genomic analysis was made applying subtractive hybridization libraries (SSH) to identify genes up-and down regulated during cooper exposure in abalone over periods of 12 and 168 h exposed to 2.5 and 10 µg/L of Cu(+2). Results obtained from the SSH library revealed 368 different sequences regulated by copper, that correspond to eight major physiological functions. The validation of these sequences obtained by SSH as well as their expression kinetics were made by PCR in real time on 14 potential genes regulated by metal stress. This study provides information for the characterization of potential genomic markers that may be used in future environmental monitoring and to investigate new mechanisms of stress to copper in this commercially important marine species.


Asunto(s)
Cobre/metabolismo , Etiquetas de Secuencia Expresada , Gastrópodos/efectos de los fármacos , Gastrópodos/genética , Contaminantes Químicos del Agua/metabolismo , Animales , Chile , Cobre/toxicidad , Monitoreo del Ambiente , Regulación de la Expresión Génica , Datos de Secuencia Molecular , Análisis de Secuencia de ADN/veterinaria , Contaminantes Químicos del Agua/toxicidad
16.
J Exp Biol ; 213(Pt 23): 4010-7, 2010 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-21075942

RESUMEN

Pesticide run-off into the ocean represents a potential threat to marine organisms, especially bivalves living in coastal environments. However, little is known about the effects of environmentally relevant concentrations of pesticides at the individual level. In this study, the suppression subtractive hybridisation technique was used to discover the main physiological function affected by a cocktail of three pesticides (lindane, metolachlor and carbofuran) in the Pacific oyster Crassostrea gigas. Two oyster populations exposed to different pollution levels in the wild were investigated. The pesticide concentrations used to induce stress were close to those found in the wild. In a time course experiment, the expression of three genes implicated in iron metabolism and oxidative stress as well as that of two ubiquitous stress proteins was examined. No clear regulation of gene or protein expression was found, potentially due to a low-dose effect. However, we detected a strong site- and organ-specific response to the pesticides. This study thus (1) provides insight into bivalve responses to pesticide pollution at the level of the transcriptome, which is the first level of response for organisms facing pollution, and (2) raises interesting questions concerning the importance of the sites and organs studied in the toxicogenomic field.


Asunto(s)
Crassostrea/efectos de los fármacos , Exposición a Riesgos Ambientales/análisis , Plaguicidas/toxicidad , Animales , Crassostrea/anatomía & histología , Crassostrea/genética , Regulación hacia Abajo/efectos de los fármacos , Francia , Perfilación de la Expresión Génica , Geografía , Proteínas de Choque Térmico/metabolismo , Hibridación de Ácido Nucleico , Especificidad de Órganos/efectos de los fármacos , Especificidad de Órganos/genética , Océano Pacífico , ARN Mensajero/genética , ARN Mensajero/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genética
17.
J Invertebr Pathol ; 105(3): 289-97, 2010 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-20692263

RESUMEN

Since 1998, episodic mass mortality of the abalone Haliotistuberculata has been observed along the northern Brittany coast of France caused by a complex interaction among the host, pathogen and environmental factors. In the present study, abalone were submitted to two successive infections with the pathogen Vibrioharveyi under controlled conditions. During the first challenge, infection by V.harveyi resulted in 64% mortality of mature abalone. After a second infection of those surviving the first challenge, only 44% mortality was observed. Physiological variability in the host response appears to be a major determinant in susceptibility to V.harveyi. In order to isolate differentially expressed genes in H.tuberculata challenged with this bacterium, suppression subtractive hybridization (SSH) cDNA libraries were constructed from muscle of moribund abalone (susceptibles), surviving individuals (apparently resistant to the bacterium) and control (unexposed) animals. Of the 1152 clones sequenced, 218 different partial cDNA sequences were obtained and represented 69 known genes. Of these, 65 were identified for the first time in H.tuberculata. Using real-time PCR, a time-course study was conducted on 19 of the genes identified by SSH. A majority of differentially expressed transcripts were down-regulated in susceptible individuals as compared to their resistant counterparts. Bacterial challenge of abalone resulted in the up-regulation of three transcripts (encoding ferritin, heat shock protein HSP84 and fatty acid binding protein FABP) in those that survived exposure to V.harveyi. This study has identified potential candidates for further investigation into the functional basis of resistance and susceptibility to summer vibriosis outbreaks in abalone.


Asunto(s)
Gastrópodos/genética , Gastrópodos/microbiología , Expresión Génica , Vibrio/fisiología , Animales , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
18.
Mar Genomics ; 3(3-4): 133-43, 2010.
Artículo en Inglés | MEDLINE | ID: mdl-21798207

RESUMEN

Marine intertidal organisms commonly face hypoxic stress during low tide emersion; moreover, eutrophic conditions and sediment nearness could lead to hypoxic phenomena; it is indeed important to understand the molecular processes involved in the response to hypoxia. In this study the molecular response of the Pacific oyster Crassostrea gigas to prolonged hypoxia (2mg O(2) L(-1) for 20d) was investigated under experimental conditions. A transcriptomic approach was employed using a cDNA microarray of 9058 C. gigas clones to highlight the genetic expression patterns of the Pacific oyster under hypoxic conditions. Lines of oysters resistant (R) and susceptible (S) to summer mortality were used in this study. ANOVA analysis was used to identify the genes involved in the response to hypoxia in comparison to normoxic conditions. The hypoxic response was maximal at day 20. The principal biological processes up-regulated by hypoxic stress were antioxidant defense and the respiratory chain compartment, suggesting oxidative stress caused by hypoxia or an anticipatory response for normoxic recovery. This is the first study employing microarrays to characterize the genetic markers and metabolic pathways responding to hypoxic stress in C. gigas.

19.
BMC Genomics ; 10: 341, 2009 Jul 29.
Artículo en Inglés | MEDLINE | ID: mdl-19640306

RESUMEN

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.


Asunto(s)
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-Computador
20.
Gene ; 442(1-2): 37-46, 2009 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-19406218

RESUMEN

Few studies have described the molecular response of mollusk larvae to heavy metal exposure. We investigated the response of Chilean scallop Argopecten purpuratus post-larvae to copper exposure under experimental conditions. Post-larvae were maintained with and without copper stress in tanks containing netlon collectors with biofilms that were formed by the bacterium Halomonas sp. and the diatom Amphora sp., known to increase larval settlement. We focused on the analysis of the differential expression patterns of genes associated with copper response. A suppression subtractive hybridization method was used to identify copper-specific up- and down-regulated genes in the post-larvae following 4 days and 8 days exposure to 2.5 and 10 microg/l Cu(+2). This method revealed 145 different sequences corresponding to 10 major physiological functions. The expression of 15 potentially regulated genes was analyzed by real-time PCR in post-larvae at different sampling times during the copper stress. The genes chosen were alpha tubulin, elongation factor 1-alpha (EF1A), tributylin binding protein type 1 (TBT), NADH dehydrogenase subunit 2 (NADH2), cavortin, glutathione peroxidase (GPx), ferritin, heat shock protein 70 (HSP70), Pam highwire rpm1 (Phr1), insulin-like growth factor 1 (IGF1), cellulase, and ribosomal proteins: L18, L44, S3a and S15. This study contributes to the characterization of potential genetic markers that could be used in future environmental monitoring and to explore new mechanisms of stress tolerance in marine mollusk species, especially in early stages of development.


Asunto(s)
Cobre/toxicidad , Monitoreo del Ambiente/métodos , Pectinidae/efectos de los fármacos , Pectinidae/genética , Animales , Perfilación de la Expresión Génica , Marcadores Genéticos , Larva/efectos de los fármacos , Larva/genética , Larva/crecimiento & desarrollo , Pectinidae/crecimiento & desarrollo , Contaminación Química del Agua
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