Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptomic responses of three-spined stickleback (Gasterosteus aculeatus) from Aire and Siblyback populations to ethinyl-estradiol, copper and binary mixtures

ABSTRACT: Sticklebacks were caught from the River Aire (Yorkshire, UK) and Siblyback reservoir (Cornwall, UK), depurated for 4 months at Exeter University, split into groups of 20 individuals before exposure. Fish were exposed to either control conditions, three concentrations of copper (3.2, 32, 128 Âµg/L; prepared using copper sulphate), three concentrations of ethinyl-estradiol (EE2) (1, 10, 32 ng/L) and three mixtures (3.2 Âµg Cu/L and 32 ng EE2/L; 128 Âµg Cu/L and 1 ng EE2/L; 128 Âµg Cu/L and 32 ng EE2/L) in duplicate tanks for 4 days. After sampling liver of male fish were stored at -80C for transcriptomics. Total RNA was prepared from fish livers, aliquots of all samples were pooled and labeled with Cy3-dCTP as a common reference, individual samples were labeled with Cy5-dCTP. Each hybridisation to the stickleback PGPS2 cDNA microarray consisited of one Cy5-labeled individual sample and one aliquot of the Cy3 labeled commmon reference sample. Data were captured with an Axon scanner using Genepix software.

INSTRUMENT(S): Axon 4000B

ORGANISM(S): Gasterosteus aculeatus

SUBMITTER: Tim Williams

PROVIDER: E-MTAB-4907 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:17alpha-ethinylestradiol (EE2) is one of the most potent estrogens that have the ability to interfere with the endocrine system of fish. The objective was to investigate the effects and mechanisms of action caused by 60 days of dietary exposure to 0.2 mg EE2/kg and 0.07 mg EE2/kg feed in female largemouth bass (LMB) during the reproductive season. Microarrays and pathway analyses were performed on hepatic tissues to identify genes and biological processes altered in female LMB by EE2 exposure. The hypothesis was that the two concentrations of EE2 would produce dose-response changes in sensitive genes. Body and ovary weights were measured and blood was collected for measurement of plasma steroid hormones (17beta-estradiol (E2), testosterone (T)) and vitellogenin (VTG) using ELISA. The 0.2 mg EE2/kg feed exposure reduced the gonadosomatic index (GSI) by 75%, and plasma levels of E2 and T were reduced by over 90%. Plasma VTG was increased by approximately 100% (from 4 to 8mg/ml) by the 0.2 mg/kg treatment. T levels, from the 0.07 mg EE2/kg feed, reduced GSI by approximately 30% and circulating E2 and T by ~80% but did not affect VTG concentrations. We found 1,594 and 1,165 genes were significantly affected (p<0.05) by the 0.07 mg EE2/kg feed and 0.2 mg EE2/kg feed, respectively. Gene ontology (GO) analysis revealed that there were different biological processes regulated by the two concentrations of EE2. Pathway analysis showed that the 0.07 mg EE2/kg feed exposure caused differential regulation of genes associated with fatty acid biosynthesis and glycolysis, indicating some metabolic effects. In contrast, the 0.2 mg EE2/kg feed exposure altered transcription of genes involved in immune response and apoptosis, suggesting a toxic response at this concentration. These results suggest that the two concentrations demonstrated distinct physiological responses, with the higher concentration inducing complete endocrine disruption in LMB. These findings demonstrate the usefulness of microarrays to identify possible biomarkers and modes of toxic action to dietary exposure in LMB. Two concentrations of EE2 would produce dose-response changes in sensitive genes. Female LMB were fed 5 days per week for 60 days with floating pellets that contained 0.07 or 0.2 mg/kg of EE2.

Dataset Information

Transcriptomic responses of three-spined stickleback (Gasterosteus aculeatus) from Aire and Siblyback populations to ethinyl-estradiol, copper and binary mixtures

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