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Using Gene Expression to Assess the Status of Fish from Anthropogenically Influenced Estuarine Wetlands

ABSTRACT: The diverse mixture of contaminants frequently present in estuarine wetlands complicates their assessment by routine chemical or biological analyses. We investigated the use of gene expression to assess contaminant exposure and the condition of southern California (USA) estuarine fish. Liver gene expression, plasma estradiol concentrations and gonad histopathology were used to investigate the biological condition of longjaw mudsuckers (Gillichthys mirabilis). A wide array of metals, legacy organochlorine pesticides, PCBs and contaminants of emerging concern were detected in sediments and whole fish. Overall gene expression patterns were characteristic to each of four sites investigated in this study. Differentially expressed genes belonged to several functional categories including xenobiotic metabolism, detoxification, disease and stress responses. In general, plasma estradiol concentrations were similar among fish from all areas. Some fish gonads had pathologic changes (e.g. infection, inflammation) that could indicate weakened immune systems and chronic stress. The differential expression of some genes involved in stress responses correlated with the prevalence of histologic gonad lesions. This study indicates that sentinel fish gene expression data is a promising tool for assessing the biological condition of fish exposed to environmental contaminants. Key Words: Gene expression, fish, contaminants, estuaries. This abstract belongs to a manuscript that has been submitted to Environmental Science and Technology. The manuscript has been invited as part of an especial Omics Issue which is expected to be published in 2012.

ORGANISM(S): Gillichthys mirabilis

PROVIDER: GSE28695 | GEO | 2011/04/19

SECONDARY ACCESSION(S): PRJNA138905

REPOSITORIES: GEO

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Project description:The diverse mixture of contaminants frequently present in estuarine wetlands complicates their assessment by routine chemical or biological analyses. We investigated the use of gene expression to assess contaminant exposure and the condition of southern California (USA) estuarine fish. Liver gene expression, plasma estradiol concentrations and gonad histopathology were used to investigate the biological condition of longjaw mudsuckers (Gillichthys mirabilis). A wide array of metals, legacy organochlorine pesticides, PCBs and contaminants of emerging concern were detected in sediments and whole fish. Overall gene expression patterns were characteristic to each of four sites investigated in this study. Differentially expressed genes belonged to several functional categories including xenobiotic metabolism, detoxification, disease and stress responses. In general, plasma estradiol concentrations were similar among fish from all areas. Some fish gonads had pathologic changes (e.g. infection, inflammation) that could indicate weakened immune systems and chronic stress. The differential expression of some genes involved in stress responses correlated with the prevalence of histologic gonad lesions. This study indicates that sentinel fish gene expression data is a promising tool for assessing the biological condition of fish exposed to environmental contaminants. Key Words: Gene expression, fish, contaminants, estuaries. This abstract belongs to a manuscript that has been submitted to Environmental Science and Technology. The manuscript has been invited as part of an especial Omics Issue which is expected to be published in 2012. In this study, we used hepatic gene expression in wild longjaw mudsuckers (Gillichthys mirabilis) to assess biological responses from anthropogenically influenced wetlands. We investigated the relationships among gene expression responses, chemical exposure and additional biological responses in this species. We studied estuarine wetlands that had diverse contaminant characteristics and received three main types of contaminant inputs in different proportions: agricultural runoff, urban runoff and municipal wastewater.

Project description:To better assess the impact of contaminants on fish reproduction, it is increasingly important to collect data at multiple levels of biological organization. However, a challenge in ecotoxicology is that three of the most commonly measured reproductive endpoints; gonad histology, ex vivo steroid production, and gene expression, can be difficult to obtain in small-bodied fish (e.g. zebrafish, Japanese medaka, fathead minnow) due to limited tissue availability. Using 5α-dihydrotestosterone (DHT) as a test compound, the objectives of the current study were to assess the feasibility of measuring gene expression in tissues also used for ex vivo steroid production studies. A portion of the testes was also used for qualitative histological examination. Male mummichog were exposed to 0, 5, and 50 µg/L DHT for 21 days. Following the exposure, testes were collected and separated into three pieces for histology, ex vivo incubation to measure steroidogenic capacity [Testosterone(T), 17β-estradiol (E2), 11-ketotestosterone (11KT)] and gene expression analyses. Testis RNA at time of dissection and testis RNA at end of ex vivo incubation (18 h) was collected and assessed for RNA integrity. The expression of genes coding for estrogen receptor βa (esrba) and estrogen receptor βb (esrbb) as well as the key steroidogenic genes, steroidogenic acute regulatory protein (star), P450 side chain cleavage (p450scc) and 11β-hydroxysteroid dehydrogenase (hsd11b3) were assessed in testis before and after ex vivo incubation. There were no significant differences in RNA quality between the two time points, however the expression of esrba, esrbb and p450scc was significantly lower post-incubation, although star, hsd11b3, and all four reference gene expression levels remained unchanged over time. DHT did not affect the expression of any of the measured genes at either concentration. Histological examination showed that all fish contained mature testes dominated by spermatozoa and spermatids. DHT did not affect T production but significantly decreased 11KT production in the high treatment group. This suggests a role for DHT in androgen regulation, and has

Project description:To better assess the impact of contaminants on fish reproduction, it is increasingly important to collect data at multiple levels of biological organization. However, a challenge in ecotoxicology is that three of the most commonly measured reproductive endpoints; gonad histology, ex vivo steroid production, and gene expression, can be difficult to obtain in small-bodied fish (e.g. zebrafish, Japanese medaka, fathead minnow) due to limited tissue availability. Using 5M-NM-1-dihydrotestosterone (DHT) as a test compound, the objectives of the current study were to assess the feasibility of measuring gene expression in tissues also used for ex vivo steroid production studies. A portion of the testes was also used for qualitative histological examination. Male mummichog were exposed to 0, 5, and 50 M-BM-5g/L DHT for 21 days. Following the exposure, testes were collected and separated into three pieces for histology, ex vivo incubation to measure steroidogenic capacity [Testosterone(T), 17M-NM-2-estradiol (E2), 11-ketotestosterone (11KT)] and gene expression analyses. Testis RNA at time of dissection and testis RNA at end of ex vivo incubation (18 h) was collected and assessed for RNA integrity. The expression of genes coding for estrogen receptor M-NM-2a (esrba) and estrogen receptor M-NM-2b (esrbb) as well as the key steroidogenic genes, steroidogenic acute regulatory protein (star), P450 side chain cleavage (p450scc) and 11M-NM-2-hydroxysteroid dehydrogenase (hsd11b3) were assessed in testis before and after ex vivo incubation. There were no significant differences in RNA quality between the two time points, however the expression of esrba, esrbb and p450scc was significantly lower post-incubation, although star, hsd11b3, and all four reference gene expression levels remained unchanged over time. DHT did not affect the expression of any of the measured genes at either concentration. Histological examination showed that all fish contained mature testes dominated by spermatozoa and spermatids. DHT did not affect T production but significantly decreased 11KT production in the high treatment group. This suggests a role for DHT in androgen regulation, and has Microarray analysis was performed with four biological replicates (4 control and 4 treatment). Male mummichog were exposed to 0, 5, and 50 M-BM-5g/L DHT for 21 days. Individuals from the high concentration, 50 ug/L DHT, were used for the microarray analysis.

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Using Gene Expression to Assess the Status of Fish from Anthropogenically Influenced Estuarine Wetlands

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