Hepatic transcriptional signatures of exposure to oestradiol (E2) and the herbicide linuron in mature male brown trout
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ABSTRACT:
ORGANISM(S): Salmo trutta
SUBMITTER: Tamsyn Uren Webster
PROVIDER: E-GEOD-57490 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:This study aimed to investigate the hepatic transcriptional response of brown trout to glyphosate, and its formulated product, Roundup. We exposed juvenile female brown trout to three concentrations of glyphosate (0.01, 0.5 and 10 mg/L) and Roundup (0.01, 0.5 and 10 mg/L glyphosate acid equivalent) for 14 days and sequenced the hepatic transcriptome of 6 individual females per treatment group in order to determine the global mechanisms of toxicity of this widely used herbicide. We assembled the brown trout transcriptome using an optimised de novo approach, and subsequent differential expression analysis identified a total of 1020 differentially-regulated transcripts across all treatments. Differentially-expressed transcripts included those encoding components of the antioxidant system, a number of stress-response proteins and pro-apoptotic signalling molecules. Functional analysis also revealed over-representation of pathways involved in regulation of cell-proliferation and turnover, and up-regulation of energy metabolism and other metabolic processes. Together, these transcriptional changes are consistent with generation of oxidative stress and the widespread induction of compensatory cellular stress response pathways. The mechanisms of toxicity identified were similar across both glyphosate and Roundup treatments, including for environmentally relevant concentrations. The significant alterations in transcript expression observed at the lowest concentrations tested raises concerns for the toxicity of this herbicide to fish populations inhabiting contaminated rivers. Fish were exposed to 3 concentrations of glyphosate (0.01, 0.1 and 10 mg/L), 3 concentrations of Roundup (0.01, 0.5 and 10 mg/L glyphosate acid equivalent) and water controls for 14 days. Liver mRNA from 6 replicate individuals per treatment was sequenced in an Illumina HiSeq 2500 platform. Two control groups (n=6 fish per group) were included. Using a de novo approach, we assembled the hepatic transcriptome for brown trout. Sequence reads were re-mapped to the assembled transcriptome using Bowtie2 and transcript expression profiling was conducted using EdgeR. ERCC spike controls were added to all individual samples, allowing for the assessment of the reproducibility and dynamic range for transcript expression quantification in our experiments. For the group exposed to 0.1 mg/L glyphosate, only 3 females were available to sequence and the variability between individuals was very high with 1 female identified as an outlier. For this reason, data from this treatment group was deemed unreliable and excluded from the analysis.
2015-04-16 | E-GEOD-56855 | biostudies-arrayexpress
Project description:These studies aimed to investigate the hepatic transcriptional response of brown trout to the natural estrogen, E2, and the herbicide linuron. We exposed mature male brown trout to three concentrations of each chemical for 4 days and sequenced the hepatic transcriptome of 3 individuals per treatment group in order to determine the global mechanisms of toxicity of these environmental contaminants. We assembled the brown trout transcriptome using a de novo approach. Subsequent differential expression analysis identified a total of 2113 differentially-regulated transcripts in the group exposed to the highest E2 treatment concentration, and 822 differentially-regulated transcripts across all linuron treatments. For E2, differentially-expressed transcripts included those encoding known oestrogen-responsive genes, while regulated processes included those associated with vitellogenesis including lipid metabolism, cellular proliferation and ribosome biogenesis. For linuron, there was a striking down-regulation of transcripts encoding the majority of the enzymes involved in the cholesterol biosynthesis pathway, and also a considerable induction of transcripts involved in cellular stress response including Cyp1a.
2015-05-08 | GSE57490 | GEO
Project description:This study aimed to investigate the hepatic transcriptional response of brown trout to glyphosate, and its formulated product, Roundup. We exposed juvenile female brown trout to three concentrations of glyphosate (0.01, 0.5 and 10 mg/L) and Roundup (0.01, 0.5 and 10 mg/L glyphosate acid equivalent) for 14 days and sequenced the hepatic transcriptome of 6 individual females per treatment group in order to determine the global mechanisms of toxicity of this widely used herbicide. We assembled the brown trout transcriptome using an optimised de novo approach, and subsequent differential expression analysis identified a total of 1020 differentially-regulated transcripts across all treatments. Differentially-expressed transcripts included those encoding components of the antioxidant system, a number of stress-response proteins and pro-apoptotic signalling molecules. Functional analysis also revealed over-representation of pathways involved in regulation of cell-proliferation and turnover, and up-regulation of energy metabolism and other metabolic processes. Together, these transcriptional changes are consistent with generation of oxidative stress and the widespread induction of compensatory cellular stress response pathways. The mechanisms of toxicity identified were similar across both glyphosate and Roundup treatments, including for environmentally relevant concentrations. The significant alterations in transcript expression observed at the lowest concentrations tested raises concerns for the toxicity of this herbicide to fish populations inhabiting contaminated rivers.
2015-04-16 | GSE56855 | GEO
Project description:Juvenile rainbow trout were exposed to two different concentrations of 17β-estradiol (E2) (2 or 20 mg/kg feed), and then infected with three concentrations of Yersinia ruckeri, a bacterial pathogen causing massive losses in wild and farmed salmonid populations. Infection with Y. ruckeri caused mortality of trout, and this effect was significantly enhanced by simultaneous exposure to high E2 dose. Analysis of hepatic gene expression profiles revealed complex regulations of pathways involved in immune responses, stress responses and detoxicification pathways. E2 markedly reduced expression of several genes implicated in xenobiotic metabolism. The results suggest that the interaction between pathogen and E2 interfered with the fish’s capability of clearing toxic compounds. The findings of the current study add to our understanding of multiple exposure responses in fish.
2012-06-17 | GSE38763 | GEO
Project description:Juvenile rainbow trout were exposed to two different concentrations of 17M-NM-2-estradiol (E2) (2 or 20 mg/kg feed), and then infected with three concentrations of Yersinia ruckeri, a bacterial pathogen causing massive losses in wild and farmed salmonid populations. Infection with Y. ruckeri caused mortality of trout, and this effect was significantly enhanced by simultaneous exposure to high E2 dose. Analysis of hepatic gene expression profiles revealed complex regulations of pathways involved in immune responses, stress responses and detoxicification pathways. E2 markedly reduced expression of several genes implicated in xenobiotic metabolism. The results suggest that the interaction between pathogen and E2 interfered with the fishM-bM-^@M-^Ys capability of clearing toxic compounds. The findings of the current study add to our understanding of multiple exposure responses in fish. Microarray analyses compared 3 groups of pathogen infected fish: no estrogen treatment (NE2), low (LE2) and high (HE2) doses of hormone
2012-06-16 | E-GEOD-38763 | biostudies-arrayexpress
Project description:Background: Previously, we reported that perfluorooctanoic acid (PFOA) promotes liver cancer in manner similar to that of 17β-estradiol (E2) in rainbow trout. Also, other perfluoroalkyl acids (PFAAs) are weakly estrogenic in trout and bind the trout liver estrogen receptor (ER). Objectives: The primary objective of this study was to determine whether multiple PFAAs enhance hepatic tumorigenesis in trout, an animal model that represents human insensitivity to peroxisome proliferators. Methods: A two-stage chemical carcinogenesis model was employed in trout to evaluate PFOA, perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluorooctane sulfonate (PFOS) and 8:2 fluorotelomer alcohol (8:2FtOH) as complete carcinogens or promoters of aflatoxin B1 (AFB1)- and/or N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced liver cancer. A custom trout DNA microarray was used to assess hepatic transcriptional response to these dietary treatments in comparison to E2 and the classic peroxisome proliferator clofibrate (CLOF). Results: Incidence, multiplicity and size of liver tumors in trout fed diets containing E2, PFOA, PFNA and PFDA were significantly higher compared to AFB1-initiated animals fed control diet, whereas PFOS caused a minor increase in liver tumor incidence. E2 and PFOA also enhanced MNNG-initiated hepatocarcinogenesis. Pearson correlation analyses, unsupervised hierarchical clustering and principal components analyses showed that the hepatic gene expression profiles for E2 and PFOA, PFNA, PFDA and PFOS were overall highly similar, though distinct patterns of gene expression were evident for each treatment, particularly for PFNA. Conclusions: Overall, these data suggest that multiple PFAAs can promote liver cancer and that the mechanism of promotion may be similar to that for E2. A total of 40 samples were analyzed using a a dye-swap, reference sample hybridization protocol. Rainbow trout were exposed to the following experimental treatments via the diet for two weeks (number in parenthesis is assigned group #): (1) Control; (2) 5 mg/kg diet estradiol (E2); (3) 2000 mg/kg diet perfluorooctanoic acid (PFOA); (4) 2000 mg/kg diet perfluorononanoic acid (PFNA); (5) 2000 mg/kg diet perfluorodecanoic acid (PFDA); (6) 200 mg/kg diet perfluorooctane sulfonate; (7) 2000 mg/kg 8:2 fluorotelomer alcohol (FTOH); and (8) 2000 mg/kg diet clofibrate. A total of 40 total hepatic mRNA samples were analyzed using a dye-swap, reference sample hybridization protocol. A reference RNA pool was made by combining equal amounts of RNA from all control RNA samples, with one exception. A separate time-matched reference pool was utilized for group 6 samples (PFOS treatment). Five hybridizations were performed for each treatment group in the following pattern: Replicate A Cy5/Reference Cy3; Replicate A Cy3/Reference Cy 5; Replicate B Cy5/Reference Cy3; Replicate B Cy3/Reference Cy5; Replicate C Cy5/Reference Cy3. Thus, the experiment consisted of three biological replicates, for two of which were replicated technically.
2011-08-01 | E-GEOD-31085 | biostudies-arrayexpress
Project description:Background: Previously, we reported that perfluorooctanoic acid (PFOA) promotes liver cancer in manner similar to that of 17β-estradiol (E2) in rainbow trout. Also, other perfluoroalkyl acids (PFAAs) are weakly estrogenic in trout and bind the trout liver estrogen receptor (ER). Objectives: The primary objective of this study was to determine whether multiple PFAAs enhance hepatic tumorigenesis in trout, an animal model that represents human insensitivity to peroxisome proliferators. Methods: A two-stage chemical carcinogenesis model was employed in trout to evaluate PFOA, perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluorooctane sulfonate (PFOS) and 8:2 fluorotelomer alcohol (8:2FtOH) as complete carcinogens or promoters of aflatoxin B1 (AFB1)- and/or N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced liver cancer. A custom trout DNA microarray was used to assess hepatic transcriptional response to these dietary treatments in comparison to E2 and the classic peroxisome proliferator clofibrate (CLOF). Results: Incidence, multiplicity and size of liver tumors in trout fed diets containing E2, PFOA, PFNA and PFDA were significantly higher compared to AFB1-initiated animals fed control diet, whereas PFOS caused a minor increase in liver tumor incidence. E2 and PFOA also enhanced MNNG-initiated hepatocarcinogenesis. Pearson correlation analyses, unsupervised hierarchical clustering and principal components analyses showed that the hepatic gene expression profiles for E2 and PFOA, PFNA, PFDA and PFOS were overall highly similar, though distinct patterns of gene expression were evident for each treatment, particularly for PFNA. Conclusions: Overall, these data suggest that multiple PFAAs can promote liver cancer and that the mechanism of promotion may be similar to that for E2.
2011-08-02 | GSE31085 | GEO
Project description:Indole-3-carbinol (I3C), from cruciferous vegetables, has been found to suppress or enhance tumors in several animal models. We previously reported that dietary I3C promotes hepatocarcinogenesis in rainbow trout (Oncorhynchus mykiss) at concentrations that differentially activated estrogen receptor (ER) or aryl hydrocarbon receptor (AhR)-mediated responses based on individual protein biomarkers. In this study, we evaluated the relative importance of these pathways as potential mechanisms for I3C on a global scale. Hepatic gene expression profiles were examined in trout after dietary exposure to 500 and 1500 ppm I3C and 3,3,-diindolylmethane (DIM), a major in vivo component of I3C, and were compared to the transcriptional signatures of two model hepatic tumor promoters; 17beta-estradiol (E2), an ER agonist, and beta-naphthoflavone, an AhR agonist. We demonstrate that I3C and DIM acted similar to E2 at the transcriptional level based on correlation analysis of expression profiles and clustering of gene responses. Of the genes regulated by E2 (fold change >2.0 or <0.50), most genes were regulated similarly by DIM (87-92%) and I3C (71%) suggesting a common mechanism of action. Of interest were upregulated genes associated with signaling pathways for cell growth and proliferation, vitellogenesis, and protein folding, stability and transport. Other genes down-regulated by E2, including those involved in acute-phase immune response, were also down-regulated by DIM and I3C. Gene regulation was confirmed by qRT-PCR and western blot. These data indicate I3C promotes hepatocarcinogenesis through estrogenic mechanisms in trout liver and suggest DIM may be an even more potent hepatic tumor promoter in this model. Keywords: dose response
2005-09-28 | GSE3324 | GEO
Project description:In this RNA-seq study, we compared the transcriptome of three Fragaria vesca genotypes in response to Phytophthora cactorum. The goal of our study was to dissect the resistance mechanism of the diploid strawberry (F. vesca) that are resistant to P. cactorum. A susceptible genotype (NCGR1218) and two resistant (NCGR1603 and Bukammen) F. vesca genotypes were used for the comparative transcriptome analyses. Plants were inoculated with P. cactorum zoospores (2mL of 2 × 105 spores/mL) in the crown (rhizome) and sampled 48 hours later. The appropriate controls for each genotype were i) samples wounded and inoculated with water and sampled 48 hours after the treatment and ii) untreated samples. Four biological replicates, each consisting of four individual test plants from each genotype were used for the transcriptome study. All the samples were collected from the crown, flash-frozen in liquid nitrogen and stored at -80 °C until RNA isolation. Total RNA was isolated using the SpectrumTM Plant Total RNA Kit (Sigma-Aldrich, USA) according to the manufacturer’s instructions. For sequencing, the libraries were prepared using the TruSeqTM stranded total RNA library prep kit (Illumina, USA), indexed and pooled, and sequenced in four lanes using the Illumina HiSeq 3/4000 (2×150 bp) System by the Norwegian Sequencing Centre, Oslo, Norway. Raw reads were quality filtered, de novo assembled into transcripts and were analysed for differentially expressed genes between the inoculated and control samples.
2023-01-01 | E-MTAB-12152 | biostudies-arrayexpress
Project description:The proliferative darkening syndrome (PDS) is an annually recurring disease that causes species-specific die-off of brown trout (Salmo trutta fario) in PDS-impacted pre-alpine rivers of central Europe. The mortality rate for PDS is near 100% and consequently the survival of brown trout populations in the impacted regions is threatened. The progression of PDS occurs in two stages, a subclinical stage and a symptomatic stage, over a time period of more than 3 months starting in spring and culminating with the die-off of brown trout in late summer. Based on experimental evidence it is hypothesized that PDS is caused by an infectious agent. To substantiate this working hypothesis as well as to discern the type of pathogen likely to be responsible for PDS, microarray analysis were conducted using a salmonid-specific cDNA microarray to assess the hepatic immune response of brown trout during the progression of PDS in 7-day intervals over a time period of 98 days.The microarray analysis revealed that brown trout suffering from PDS exhibit increased hepatic expression of important anti-viral genes both during the subclinical stage of PDS, namely the Barrier-to-autointegration factor, and during the symptomatic stage of PDS, namely the interferon regulatory factor 1 as well as the Guanylate-binding protein 1. Additionally, during the symptomatic stage of PDS there is strong hepatic up-regulation of the chemokine CCL19, complement components C1QC and C6, Proteasome activator complex subunits 1 and 2 as well as a variety of both MHC class I and MHC class II transcripts. In conclusion, the increased expression of hepatic immune response genes involved in a variety of immune system processes that mediate defense against infection identified by the microarray analysis during the progression of PDS support the working hypothesis that PDS is caused by an infectious agent. Furthermore, the up-regulation of antiviral immune response genes during both the subclinical stage and the symptomatic stage of PDS suggests that this disease is caused by a pathogenic virus. On May 29, 2008, brown trout (Salmo trutta fario) of the same age class (1+) with an individual weight ranging between 25-85 grams were obtained from a single hatchery (Schwäbischer Fischereihof Salgen, Fachberatung für Fischerei Schwaben, Germany) and randomly allocated to one of two different experimental stations that are both located along the Iller river, named here simply the control station (location by Oberstdorf, Germany) and the treatment station (location by Kempten, Germany). At both experimental stations brown trout were held in tanks (n=750 at the treatment station and n=70 at the control station) that were supplied with water from the Iller river in a flow-through system. Sampling at both experimental stations started on May 29, 2008, which was also the day on which the fish were first transferred to their exposure tanks (referred to as 0 day post exposure; d.p.e), and ended on the 5th of September 2008. At the treatment station 3 fish were sampled each day (always at 2pm), whereas at the control station 3 fish were sampled in 7-day intervals (always at 12 noon). Fish were anaesthetized by a blow to the head and organ tissue of interest (liver, kidney, spleen, gill, muscle, stomach and foregut tissue) were immediately harvested from sacrificed fish, snap-frozen in liquid nitrogen and subsequently stored at -80°C. Livers from the three brown trout (n=3) that were sampled concurrently from both treatment and control group (n=2) in 7-day intervals starting 7 d.p.e (7, 14, 21, 28, 35, 42, 49, 56, 63, 70, 77, 84, 91 and 98 d.p.e; n=14) were recruited for the use in the microarray analysis. Microarray analyses were performed using a direct comparison two-channel design in which equimolar amounts of liver sample of one brown trout from both treatment and control group were co-hybridized on the same microarray. For each time point (n=14) microarray co-hybridizations were repeated in triplicate (n=3) and included one dye-swap in order to reduce dye-bias. Thus a total of 42 microarray slides were used in this study (3 biological replicate microarrays x 14 time points = 42 microarrays).
2015-06-26 | E-GEOD-70257 | biostudies-arrayexpress