Unknown,Transcriptomics,Genomics,Proteomics

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HEPATIC GENE EXPRESSION PATTERNS IN JUVENILE TURBOTS (Scophthalmus maximus) AFTER EXPOSURE TO STYRENE (PRAGMA, a pragmatic and integrated approach for the evaluation of environmental impact of oil and chemical spills at sea)

ABSTRACT: Accidental spills of chemical or oil tankers such as Levoli Sun or Prestige have recently impacted the European seas. The analysis of the genes regulated by exposure to spilled chemical substances could help elucidating adaptatory and compensatory pathways that may participate in pathogenesis in marine biota. Hepatic expression profiles of male juvenile S. maximus exposed to a nÂº2 fuel-oil (FO) and to styrene were studied employing a turbot custom microarray containing 2715 3â-UTR biassed single-sequences, many of them unannotated. Hybridisations identified 169 and 293 genes significantly regulated after 3 and 14 days of exposure to FO, respectively. Typical PAH responsive genes such as AhRR, ARNT2, CYP1A1 and GST were significantly upregulated. Exposure significantly affected genes involved in protein metabolism, synthesis being the most regulated pathway with ribosomal proteins and various translation factors overexpressed. Protein synthesis could participate in compensatory pathways after FO induced protein damage. Additionally, two weeks of exposure resulted in the upregulation of ras pathway genes. Regarding styrene, gene expression was very variable within experimental groups and only 2 genes were significantly regulated after 3 days of exposure. After 1 week of exposure styrene modified the expression levels of 44 genes. Most highly regulated genes, that may constitute new biomarkers of exposure to styrene, included those coding for estrogen-responsive finger protein, transmembrane 9 superfamily member 4 or selenoprotein-T, but the only pathway found to be significantly regulated was once again that of protein synthesis. Funded EU project PRAGMA, Spanish MEC (CANCERMAR), Basque Government (Consolidated Research Groups GIC07/26-IT-393-07). Turbots were exposed to styrene and to seawater control. After 3 and 7 days of exposure turbots were in situ dissected and liver was extracted, inmersed in RNAlater and frozen in liquid N2

ORGANISM(S): Scophthalmus maximus

SUBMITTER: Ibon Cancio

PROVIDER: E-GEOD-43359 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Accidental spills of chemical or oil tankers such as Levoli Sun or Prestige have recently impacted the European seas. The analysis of the genes regulated by exposure to spilled chemical substances could help elucidating adaptatory and compensatory pathways that may participate in pathogenesis in marine biota. Hepatic expression profiles of male juvenile S. maximus exposed to a nÂº2 fuel-oil (FO) and to styrene were studied employing a turbot custom microarray containing 2715 3â-UTR biassed single-sequences, many of them unannotated. Hybridisations identified 169 and 293 genes significantly regulated after 3 and 14 days of exposure to FO, respectively. Typical PAH responsive genes such as AhRR, ARNT2, CYP1A1 and GST were significantly upregulated. Exposure significantly affected genes involved in protein metabolism, synthesis being the most regulated pathway with ribosomal proteins and various translation factors overexpressed. Protein synthesis could participate in compensatory pathways after FO induced protein damage. Additionally, two weeks of exposure resulted in the upregulation of ras pathway genes. Regarding styrene, gene expression was very variable within experimental groups and only 2 genes were significantly regulated after 3 days of exposure. After 1 week of exposure styrene modified the expression levels of 44 genes. Most highly regulated genes, that may constitute new biomarkers of exposure to styrene, included those coding for estrogen-responsive finger protein, transmembrane 9 superfamily member 4 or selenoprotein-T, but the only pathway found to be significantly regulated was once again that of protein synthesis. Funded EU project PRAGMA, Spanish MEC (CANCERMAR), Basque Government (Consolidated Research Groups GIC07/26-IT-393-07). Turbots were exposed to fuel-oil and to seawater control: 7 turbots in Control tank and 8 in Oiled tank. After 3 days of exposure turbots were in situ dissected and liver was extracted, inmersed in RNAlater and frozen in liquid N2

Project description:Accidental spills of chemical or oil tankers such as Levoli Sun or Prestige have recently impacted the European seas. The analysis of the genes regulated by exposure to spilled chemical substances could help elucidating adaptatory and compensatory pathways that may participate in pathogenesis in marine biota. Hepatic expression profiles of male juvenile S. maximus exposed to a nÂº2 fuel-oil (FO) and to styrene were studied employing a turbot custom microarray containing 2715 3â-UTR biassed single-sequences, many of them unannotated. Hybridisations identified 169 and 293 genes significantly regulated after 3 and 14 days of exposure to FO, respectively. Typical PAH responsive genes such as AhRR, ARNT2, CYP1A1 and GST were significantly upregulated. Exposure significantly affected genes involved in protein metabolism, synthesis being the most regulated pathway with ribosomal proteins and various translation factors overexpressed. Protein synthesis could participate in compensatory pathways after FO induced protein damage. Additionally, two weeks of exposure resulted in the upregulation of ras pathway genes. Regarding styrene, gene expression was very variable within experimental groups and only 2 genes were significantly regulated after 3 days of exposure. After 1 week of exposure styrene modified the expression levels of 44 genes. Most highly regulated genes, that may constitute new biomarkers of exposure to styrene, included those coding for estrogen-responsive finger protein, transmembrane 9 superfamily member 4 or selenoprotein-T, but the only pathway found to be significantly regulated was once again that of protein synthesis. Funded EU project PRAGMA, Spanish MEC (CANCERMAR), Basque Government (Consolidated Research Groups GIC07/26-IT-393-07). Turbots were exposed to fuel-oil and to seawater control: 7 turbots in Control tank and 8 in Oiled tank. After 14 days of exposure turbots were in situ dissected and liver was extracted, inmersed in RNAlater and frozen in liquid N2

Project description:Pharmaceutical chemicals used in human medicine are released into surface waters via municipal effluents and pose a risk for aquatic organisms. Among these substances are selective serotonin reuptake inhibitors (SSRIs) which can affect aquatic organisms at sub ppb concentrations. To better understand biochemical pathways influenced by SSRIs, evaluate changes in the transcriptome, and identify gene transcripts with potential for biomarkers of exposure to SSRIs; larval zebrafish Danio rerio were exposed (96 h) to two concentrations (25 and 250 µg/L) of the SSRIs, fluoxetine and sertraline, and changes in global gene expression were evaluated (Affymetrix GeneChip® Zebrafish Array). Significant changes in gene expression (>=1.7 fold change, p<0.05) were determined with Partek® Genomics Suite Gene Expression Data Analysis System and ontology analysis was conducted using Molecular Annotation System 3. The number of genes differentially expressed after fluoxetine exposure was 288 at 25 µg/L and 131 at 250 µg/L; and after sertraline exposure was 33 at 25 µg/L and 52 at 250 µg/L. Five genes were differentially regulated in all treatments relative to control, suggesting that both SSRIs share some similar molecular pathways. Among them, expression of the gene coding for FK506 binding protein 5 (FKBP5), which is annotated to stress response regulation, was highly down-regulated in all treatments (results confirmed by qRT-PCR). Gene ontology analysis indicated that regulation of stress response and cholinesterase activity were critical functions influenced by these SSRIs, and suggested that changes in the transcription of FKBP5 or acetylcholinesterase could be useful biomarkers of SSRIs exposure in wild fish. Zebrafish (Danio rerio) were obtained from the Zebrafish Research Facility maintained at the Center for Environmental Biotechnology at the University of Tennessee. Fish husbandry, spawning, and experimental procedures were conducted with approval from the UT Insititutional Animal Care and Use Committee (Protocol #1690-1007). Water for holding fish and conducting experiments (hereafter referred to as fish water) consisted of MilliQ water (Millipore, Bedford, MA) with ions added: 19 mg/L NaHCO3, 1 mg/L sea salt (Instant Ocean Synthetic Sea Salt, Mentor, OH), 10 mg/L CaSO4, 10 mg/L MgSO4, 2 mg/L KCl. Embroyos were obtained by spawning adult fish with no history of contaminant exposure. Fertilization of embryos took place at the same time (<15 minutes), such that larvae used in experiments were of similar age at the time of exposure. All activities (maintenance of adult fish, spawning, and experiments) were conducted in an environmental chamber with a temperature of 27 +/- 1 C and 14:10h light:dark photoperiod. Larval zebrafish (72 hpf) were exposed for 96 h in 200ml fish water containing appropirate amount of SSRI stock (i.e. fluoxetine or sertraline). There were four SSRIs treatments (25 and 250 ug/L fluoxetine and 25 and 250 ug/L sertraline) and one control (no SSRIs) with triplicate beakers and each beaker contained about 100 larval fish. During exposure for 96 hours, beakers were kept covered to prevent water evaporation and fish were not fed (i.e., fish consumed their yolk sac).

Project description:Zebrafish (Danio rerio) were obtained from the Zebrafish Research Facility maintained in the Center for Environmental Biotechnology at the University of Tennessee. Fish husbandry, spawning, and experimental procedures were conducted with approval from the University of Tennessee Institutional Animal Care and Use Committee (Protocol #1690-1007). Water for holding fish and conducting experiments (hereafter referred to as fish water) consisted of MilliQ water (Millipore, Bedford, MA) with ions added: 19 mg/L NaHCO3, 1 mg/L sea salt (Instant Ocean Synthetic Sea Salt, Mentor, OH), 10 mg/L CaSO4, 10 mg/L MgSO4, 2 mg/L KCl. Embryos were obtained by spawning adult fish with no history of contaminant exposure. Fertilization of embryos took place at the same time (Â± 15 min.), such that larvae used in experiments were of similar age at the time of exposure. All activities (maintenance of adult fish, spawning, and experiments) were conducted in an environmental chamber with a temperature of 27Â± 1 ÂºC and 14:10h light:dark photoperiod. Experiment Overall Design: At 72 h post-fertilization, zebrafish larvae were exposed to lyophilized Microcystis and purified MC-LR at concentrations of 100 and 1,000 Âµg/L. Controls consisted of zebrafish system water (negative control) and zebrafish system water containing 0.05% ethanol (vehicle control). Larvae from both control groups as well as 100 Âµg/L MC-LR, 1,000 Âµg/L MC-LR, and lyophilized Microcystis were exposed in groups of 50 with three replicates and were sacrificed after 96 hours for total RNA extraction and subsequent microarray analysis. All larvae were exposed in beakers containing 100 ml of solution. Experiment Overall Design: Water samples for microcystin analysis and water quality measurements were taken during the experiment, and mortality and behavioral observations were recorded at 24-hour intervals. Microcystin analysis was conducted by protein phosphatase inhibition assay. Measured concentrations of microcystin-LR were 140 Â± 12 SD (low concentration) and 1,703 Â± 71 SD (high concentration). The concentration of microcystin-LR in the lyophilized Microcystis treatment was 4.5 Âµg/L. Water quality parameters measured included dissolved oxygen (6.7 mg/L), pH (6.9), total alkalinity (36 mg/L as CaCO3), total hardness (18 mg/L as CaCO3), and ammonia (<0.2 mg/L). No significant mortality or behavioral changes in larvae were observed during the exposure.

Dataset Information

HEPATIC GENE EXPRESSION PATTERNS IN JUVENILE TURBOTS (Scophthalmus maximus) AFTER EXPOSURE TO STYRENE (PRAGMA, a pragmatic and integrated approach for the evaluation of environmental impact of oil and chemical spills at sea)

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