A DNA expression array to detect toxic stress response in European flounder (Platichthys flesus)

Williams, Tim D.; Gensberg, Karl; Minchin, Stephen; Chipman, J.K.

doi:10.1016/s0166-445x(03)00119-x

Cited by 208 publications

(124 citation statements)

References 66 publications

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“…The application of gene array technology to study effects of environmental contaminants and hypoxia in aquatic systems is in the early stages of development and has been limited to fish (Gracey et al 2001, Hogstrand et al 2001, Larkin et al 2003, Ton et al 2003, Williams et al 2003. The identification of hypoxia-responsive genes and proteins of the blue crab in this study constitutes an important first step toward a better understanding of the benefits and problems associated with the use of gene and protein expression profiles in estuarine invertebrates as molecular indicators of environmental conditions.…”

Section: Discussionmentioning

confidence: 99%

Molecular indicators of hypoxia in the blue crab Callinectes sapidus

Brown‐Peterson¹,

Larkin²,

Denslow³

et al. 2005

Mar. Ecol. Prog. Ser.

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Occurrence of hypoxia in estuarine waters is increasing, and recovery of impacted estuaries is slow. Detection of early effects of hypoxia is needed for timely remedial action to be taken. Here we examine the use of hypoxia-responsive gene and protein expression profiles in the blue crab Callinectes sapidus as early indicators of impacts of hypoxia. We cloned 23 potential hypoxiaresponsive genes, which were used to construct gene macroarrays. Crabs exposed to chronic hypoxia (2.5 ppm dissolved oxygen [DO]) for 15 d showed significant (p < 0.05) changes in gene expression of heat shock protein 70 (Hsp70), copper metallothionein (CuMt3), cytosolic MnSOD (cyt-MnSOD) and ribosomal proteins S15 and L23. In all cases except for CuMt3, gene expression decreased after 5 d exposure to hypoxia. Expression of Hsp70, CuMt3 and cyt-MnSOD also increased (p < 0.05) in normoxic crabs held for 15 d, suggesting confounding effects from confinement stress. Hemocyanin protein concentrations changed significantly (p = 0.005) across the 15 d chronic hypoxia exposure. Hemocyanin in crabs exposed to 10 d intermittent hypoxia (2.5 to 8 ppm DO over a 24 h cycle) did not change, but cyt-MnSOD gene expression increased significantly (p = 0.037), whereas cytochrome c oxidase subunit 1 (ccox1) showed a 2.2-fold downregulation. Blue crabs collected from Pensacola Bay, Florida, showed significant (p < 0.006) downregulation of ccox1 and cyt-MnSOD gene expression as well as hemocyanin protein levels at a diurnally hypoxic marsh site. Several hypoxiaresponsive genes (elongation factor 2, cryptocyanin, hemocyanin) were also significantly elevated (p < 0.006) in intermolt versus premolt normoxic crabs. The identification of hypoxia-responsive genes and proteins in the blue crab is a promising first step towards the development of sensitive molecular tools for the detection of sublethal effects of hypoxia in estuarine-resident species.KEY WORDS: Dissolved oxygen · Hypoxia · Crustacea · Molecular biomarker · Gene arrays Resale or republication not permitted without written consent of the publisher

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Section: Discussionmentioning

confidence: 99%

Molecular indicators of hypoxia in the blue crab Callinectes sapidus

Brown‐Peterson¹,

Larkin²,

Denslow³

et al. 2005

Mar. Ecol. Prog. Ser.

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“…This high-throughput genome analysis can be applied for studying DNA-protein interaction, transcription analysis, detection and characterization of genetic variants [43] and environmental toxicology [44]. The summary of microarray analysis is shown in Fig.…”

Section: Introduction To Microarray Technologymentioning

confidence: 99%

Microarray analysis of metallothioneins in human diseases—A review

Křížková

Kępińska

Emri

et al. 2016

Journal of Pharmaceutical and Biomedical Analysis

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. 1 Microarray analysis of metallothioneins in human diseases AbstractMetallothioneins (MTs), low molecular mass cysteine-rich proteins, which are able to bind up to 20 monovalent and up to 7 divalent heavy metal ions are widely studied due to their functions in detoxification of metals, scavenging free radicals and cells protection against the oxidative stress. It was found that the loss of the protective effects of MT leads to an escalation of pathogenic processes and carcinogenesis. 3The most extensive area is MTs expression for oncological applications, where the information about gene patterns is helpful for the identification biological function, resistance to drugs and creating the correct chemotherapy. In other medical applications the effect of oxidative stress to cell lines exposed to heavy metals and hydrogen peroxide is studied as well as influence of drugs and cytokines on MTs expression and MTs expression in the adiposetissue. The precise detection of low metallothionein concentrations and its isoforms is necessary to understand the connection between quantity and isoforms of MTs to size, localization and type of cancer. This information is necessary for well-timed therapy and increase the chance to survival. Microarray chips appear as good possibility for finding all information about expression of MTs genes and isoforms not only in cancer, but also in other diseases, especially diabetes, obesity, cardiovascular diseases, ageing, osteoporosis, psychiatric disorders and as the effects of toxic drugs and pollutants, which is discussed in this review. List of abbreviations:HCC -hepatocellular carcinoma, HFD -high fat diet, MMP -matrix metalloproteinase, MT -metallothionein, MTF -metal transcription factor, MRE -metal responsive element, ROS -reactive oxygen species.

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“…Transcriptomic profiling with targeted microarrays has been used to assess the effects of exposure to polluted sediments and in some of these studies it has been possible to obtain a toxicologically relevant signal from amongst the transcripts which were changed. Thus, when comparing a highly PAH-contaminated environment with a 'clean' reference site, flounder transcripts characteristic of acute PAH-response were changed (Williams et al, 2003). Similarly, when comparing hornyhead turbot (Pleuronichthys verticalis) from a site receiving an estrogenic effluent with a clean site, a signal characteristic of acute estrogen exposure was observed (Baker et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Hepatic gene expression in flounder chronically exposed to multiply polluted estuarine sediment: Absence of classical exposure ‘biomarker’ signals and induction of inflammatory, innate immune and apoptotic pathways

et al. 2010

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The effects of chronic long term exposure to multiply-polluted environments on fish are not well understood, but environmental surveys suggest that such exposure may cause a variety of pathologies, including cancers. Transcriptomic profiling has recently been used to assess gene expression in European flounder (Platichthys flesus) living in several polluted and clean estuaries. However, the gene expression changes detected were not unequivocally elicited by pollution, most likely due to the confounding effects of natural estuarine ecosystem variables. In this study flounder from an uncontaminated estuary were held on clean or polluted sediments in mesocosms, allowing control of variables such as salinity, temperature, and diet. After 7 months flounder were removed from each mesocosm and hepatocytes prepared from fish exposed to clean or polluted sediments. The hepatocytes were treated with benzo(a)pyrene (BAP), estradiol (E2), copper, a mixture of these three, or with the vehicle DMSO. A flounder cDNA microarray was then used to measure hepatocyte transcript abundance after each treatment. The results show that long term chronic exposure to a multiply-polluted sediment causes increases in the expression of mRNAs coding for proteins of the endogenous apoptotic program, of innate immunity and inflammation. Contrary to expectation, the expression of mRNAs which are commonly used as biomarkers of environmental exposure to particular contaminants were not changed, or were changed contrary to expectation. However, acute treatment of hepatocytes from flounder from both clean and polluted sediments with BAP or E2 caused the expected changes in the expression of these biomarkers. Thus transcriptomic analysis of flounder exposed long-term to chronic pollution causes a different pattern of gene expression than in fish acutely treated with single chemicals, and reveals novel potential biomarkers of environmental contaminant exposure. These novel biomarkers include Diablo, a gene involved in apoptotic pathways and highly differentially regulated by both chronic and acute exposure to multiple pollutants.

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A DNA expression array to detect toxic stress response in European flounder (Platichthys flesus)

Cited by 208 publications

References 66 publications

Molecular indicators of hypoxia in the blue crab Callinectes sapidus

Molecular indicators of hypoxia in the blue crab Callinectes sapidus

Microarray analysis of metallothioneins in human diseases—A review

Hepatic gene expression in flounder chronically exposed to multiply polluted estuarine sediment: Absence of classical exposure ‘biomarker’ signals and induction of inflammatory, innate immune and apoptotic pathways

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