Dataset Information

Assessing the Potential Ecological Impacts of Perfluorinated Chemicals (PFCs) in the liver and testis of male largemouth bass (Micropterus salmoides) in Minnesota Lakes

ABSTRACT: This study used an emerging analytical technology (cDNA microarrays) to assess the potential effects of PFC exposure on largemouth bass in TCMA lakes. Microarrays simultaneously measure the expression of thousands of genes in various tissues from organisms exposed to different environmental conditions. From this large data set, biomarkers (i.e., genes that are expressed in response to an exposure to known stressors) and bioindicators (e.g., suites of genes that correspond to changes in organism health) can be simultaneously measured to clarify the relationship between contaminant exposure and organism health. Based on current scientific literature, we hypothesized that gene expression patterns would be altered in fish exposed to PFCs (as compared with fish from reference lakes), and that the magnitude of these changes would correspond to the concentrations of PFCs present throughout TCMA lakes. Patterns of gene expression in largemouth bass observed across the TCMA lakes corresponded closely with PFC concentration. Concentrations of PFCs in largemouth bass varied significantly across the sampled lakes, where the lowest concentrations were found in Steiger and Upper Prior Lakes and the highest concentrations were found in Calhoun and Twin Lakes. Patterns of gene expression were most different (relative to controls) in fish with the highest PFC tissue concentrations, where fish from Twin and Calhoun Lakes were observed to have between 5437 and 5936 differentially expressed genes in liver and gonad tissues. Although gene expression patterns demonstrated a high degree of correlation with PFC concentrations, microarray data also suggest there are likely additional factors influencing gene expression patterns in largemouth bass in TCMA lakes.

ORGANISM(S): Micropterus salmoides

PROVIDER: GSE57684 | GEO | 2014/07/31

SECONDARY ACCESSION(S): PRJNA247676

REPOSITORIES: GEO

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Project description:The NationM-bM-^@M-^Ys streams and rivers contain several contaminants in the form of complex mixtures. These cocktails of chemicals are not equivalent in concentrations, some pollutants such as nutrients can be found in the range of mg/L (macro-pollutants) but others components (micro-pollutants) such as endocrine disrupting chemicals (EDCs) are in amounts thousands to millions of times less concentrated ug/L to ng/L. These mixtures hamper the determination of particular effects of contaminants in aquatic biota. Nonetheless, the fact that toxicity is preceded by alteration in gene expression in an organism allows the use of gene expression profiling (from microarray studies) to detect early toxic effects and identify mechanisms of action. The microarray technology, a collection of DNA fragments attached to a solid surface, can be used to measure the expression levels of large numbers of genes. This facilitates establishment of links between toxicants and effects on biota. In urban waters, micropollutants such as EDCs, are known to cause effects at very low concentrations. One common class of EDCs found in low levels in urban waters is the class of perfluorochemicals (PFCs). Previously, we observed that urban waters with wastewater influence containing PFCs in the 300 ng/L range exerted effects in fish by altering the expression of cholesterol metabolism and DNA repair genes in the liver. To determine whether low concentrations in the range of the PFCs found in the environment can elicit gene expression changes, we investigated the impact of 7 different types of PFCs in a controlled laboratory study by exposing fathead minnows for 48h to environmentally relevant concentrations of PFCs. Additionally, we use blood as starting material for microarray analysis in order to explore non-invasive techniques. No fish mortality was observed in any treatment exposures, but gene expression was altred. Surprisingly, low levels of PFCs that we used altered gene expression in fish liver and blood. Several of the same genes were altered in both liver and blood from exposed fish. Micorarray analysis yields information on altered molecular pathways that predict changes at higher levels of biological organization such as survival and reproduction. Gene expression after 48 hours of exposure to prefluorochemicals in male liver of FHM Exposure Water Preparation Four different treatment were prepared. A control (0 ug/L); two PFOS concentrations: PFOS high (25 ug/L), PFOS low (0.5 ug/L); and a PFCs mixture, consisting in 7 types of PFCs at concentrations similar than those found in a previous research downstream to a wastewater treatment plant (Rodriguez-Jorquera et al., in prep.) in an attempt to mimic the mixtures usually found in waters with wastewater influence. The preparation of all exposure treatments was completed dissolving the needed amount of PFCs for each concentration first on a 50 ml polypropylene falcon tube and 1, 9 ul of TEG (Triethylene glycol) as vehicle. For the control group, Milli-Q water and TEG was mixed in a falcon tube. Then, the 50 ml preparation were poured into a pre cleaned fiberglass cylinder containing 38 liters of carbon filtered and de-chlorinated city water. All PFCs were purchased from Wellington labs. Water samples were collected at the end of the exposures from the distribution cylinders. 1 L sample was collected for each treatment in a polypropylene bottles for PFCs analysis of 7 types of perfluorinated carboxylic acids (including PFOA) using EPA Method 537. Fish Exposure and Tissue Collection Thirty two males were separated from the common tank two weeks before the experiment and placed in the treatment aquaria for 48 h. The exposure system consisted of 10 L glass aquaria. Each exposure was conducted in quadruplicate and each aquarium contained two male FHM in 4 L of treatment water. The water used in the control treatment was carbon filtered, dechlorinated tap water. The positions of the treatment tanks were randomized and test initiation times were staggered to ensure an exposure/sampling interval of 48 h. The fish were not fed during the experiment. The temperature range of the water was 24- 26 M-BM-0C with a photoperiod of 16 h light: 8 h dark. At the conclusion of the exposures, fish were anesthetized with MS-222 and weighed to the nearest 0.1 g .The testes were removed and preserved for histological analysis to check sex and sexual stage. Liver tissue and whole blood was flash frozen using liquid nitrogen and stored at -80 M-BM-0C until RNA extraction. Liver and blood were isolated from all males. The liver and blood from same individuals (four for each treatment) were used for RNA extraction. All procedures involving live fish were reviewed and approved by the University of Florida Institutional Animal Care and Use Committee (IACUC). After the exposure of fish to each treatment, four biological replicates were used to isolate RNA from FHM livers and blood using the same individuals in order to diminish variability among fish.Then microarrays were washed according to the Agilent protocol and, kept in the dark until scanning.

Project description:The Nation’s streams and rivers contain several contaminants in the form of complex mixtures. These cocktails of chemicals are not equivalent in concentrations, some pollutants such as nutrients can be found in the range of mg/L (macro-pollutants) but others components (micro-pollutants) such as endocrine disrupting chemicals (EDCs) are in amounts thousands to millions of times less concentrated ug/L to ng/L. These mixtures hamper the determination of particular effects of contaminants in aquatic biota. Nonetheless, the fact that toxicity is preceded by alteration in gene expression in an organism allows the use of gene expression profiling (from microarray studies) to detect early toxic effects and identify mechanisms of action. The microarray technology, a collection of DNA fragments attached to a solid surface, can be used to measure the expression levels of large numbers of genes. This facilitates establishment of links between toxicants and effects on biota. In urban waters, micropollutants such as EDCs, are known to cause effects at very low concentrations. One common class of EDCs found in low levels in urban waters is the class of perfluorochemicals (PFCs). Previously, we observed that urban waters with wastewater influence containing PFCs in the 300 ng/L range exerted effects in fish by altering the expression of cholesterol metabolism and DNA repair genes in the liver. To determine whether low concentrations in the range of the PFCs found in the environment can elicit gene expression changes, we investigated the impact of 7 different types of PFCs in a controlled laboratory study by exposing fathead minnows for 48h to environmentally relevant concentrations of PFCs. Additionally, we use blood as starting material for microarray analysis in order to explore non-invasive techniques. No fish mortality was observed in any treatment exposures, but gene expression was altred. Surprisingly, low levels of PFCs that we used altered gene expression in fish liver and blood. Several of the same genes were altered in both liver and blood from exposed fish. Micorarray analysis yields information on altered molecular pathways that predict changes at higher levels of biological organization such as survival and reproduction.

Dataset Information

Assessing the Potential Ecological Impacts of Perfluorinated Chemicals (PFCs) in the liver and testis of male largemouth bass (Micropterus salmoides) in Minnesota Lakes

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