Unknown,Transcriptomics,Genomics,Proteomics

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Transcript profiles of juvenile female brown trout exposed to glyphosate and Roundup herbicides.

ABSTRACT: 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.

ORGANISM(S): Salmo trutta

SUBMITTER: Tamsyn Uren Webster

PROVIDER: E-GEOD-56855 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Glyphosate-based herbicides (GBH) are the major pesticides used worldwide. Converging evidence suggests that GBH residues pose a particular risk to the kidneys and liver. However, the existence of biological effects with negative health implications at low environmentally relevant doses remains unresolved. A previous investigation addressed this issue, by conducting a 2-year feeding study, which included 10 female Sprague Dawley rats administered via drinking water with 0.1 ppb of a major Roundup formulation (50 ng/L glyphosate equivalent dilution). Hepatorenal toxicities, as well as urine and blood biochemistry disturbances at the 15th month of age were observed. In an effort to obtain molecular mechanistic insight into the underlying causes of these pathologies, we have carried out a transcriptome microarray analysis of the liver and kidneys from these same animals. The expression of 4224 and 4447 genes were found to be disturbed respectively in liver and kidney (p<0.01, q<0.08, fold change >1.1). Among the 1319 genes whose expression was altered in both tissues, 3 functional categories were over-represented. First, genes involved in mRNA splicing and small nucleolar RNA were mostly upregulated, suggesting disruption of normal spliceosome activity. Electron microscopic analysis of hepatocytes confirmed nucleolar structural disruption. Second, genes controlling chromatin structure (especially histone-lysine N-methyltransferases) were mostly upregulated. Third, genes related to respiratory chain complex I and the tricarboxylic acid cycle were mostly downregulated. The transcription factor networks that can account for these disruptions were centered on CREB1, ESR1, YY1, c-Myc and Oct3/4 activity, which are known to closely cooperate in the regulation of gene expression after hormonal stimulation. The analysis of pathways and toxicity processes showed that these disturbances in gene expression were representative of fibrosis, necrosis, phospholipidosis, mitochondrial membrane dysfunction and ischemia, which correlate with the pathologies observed at an anatomical and histological level. Our results suggest that new studies incorporating testing principles from endocrinology and developmental epigenetics need to be performed to investigate potential consequences of exposure to low dose, environmental levels of GBH and glyphosate. Nine female rats were fed drinking water containing 0.1ppb Roundup formulation (50 ng/L glyphosate equivalent dilution). Ten control female rats were fed untreated drinking water. Samples of liver and kidney were taken for gene expression analysis.

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Transcript profiles of juvenile female brown trout exposed to glyphosate and Roundup herbicides.

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