Seonock Woo

Effects of heavy metals on antioxidants and stress-responsive gene expression in Javanese medaka (Oryzias javanicus).

The differential expression of eight genes encoding stressor biomarkers was investigated by real-time quantitative PCR in liver tissue extracted from Javanese medaka after exposure to six heavy metals for 24 h. OjaCAT transcription increased in a dose-dependent manner during exposure to Cd, Cu, and Zn, but significantly decreased after exposure to Ag, Cr, and Ni. OjaCYP1A transcription decreased drastically on exposure to all heavy metals tested. OjaG6PD transcription increased dramatically after exposure to low doses of Cu and Zn, but decreased at high concentrations of these elements. No prominent changes in OjaG6PD transcription were observed after exposure to Ag, Cd, Cr, or Ni. OjaGPx mRNA expression was induced in the liver following exposure to Ag, Cd, Cu, and Zn, but suppressed following exposure to Cr and Ni. Exposure to all heavy metals increased transcription of OjaGR and OjaGST in a dose-dependent manner. OjaSOD transcription increased during exposure to Ag, Cd, Zn, and Cr, but showed no change in response to Cu and Ni exposure. OjaUB expression was induced by all doses of exposure. The transcriptional responses of these genes to heavy metal exposure will provide the basis for a multi-biomarker system that can be used for the biomonitoring of aquatic environments.

Transcripts level responses in a marine medaka (Oryzias javanicus) exposed to organophosphorus pesticide.

The differential expression of a set of genes encoding antioxidant enzymes and stress-responsive proteins was investigated by real-time quantitative PCR in intestine, liver, and muscle tissues extracted from Oryzias javanicus after exposure to the organophosphorus pesticide, Iprobenfos (IBP). After IBP exposure, transcriptional changes in all the tested genes were prominent in the liver and moderate in the intestine, but unpredictable in the muscle. In the liver, CAT transcription increased after exposure to IBP at all concentrations (P<0.05). CYP1A mRNA was induced in the intestine and liver at the two higher concentrations. G6PD transcription was induced in the liver at the three higher IBP concentrations, but was suppressed in muscle at the same concentrations. GPx expression in the liver increased at three higher concentrations of IBP. In the intestine and liver, GR expression was induced at two higher and three higher concentrations, respectively. However, no significant changes were observed in the muscle. GST and SOD transcription was induced in the liver at all IBP concentrations. IBP exposure induced UB expression in the intestine and liver in a concentration-dependent manner. The transcriptional changes in these genes in the liver could be good biomarkers for stress levels in O.javanicus, and be used as critical biomarkers for environmental quality assessment.

Differentially displayed genes with oxygen depletion stress and transcriptional responses in the marine mussel, Mytilus galloprovincialis

Hypoxic events affecting aquatic environments have been reported worldwide and the hypoxia caused by eutrophication is considered one of the serious threats to coastal marine ecosystems. To investigate the molecular-level responses of marine organisms exposed to oxygen depletion stress and to explore the differentially expressed genes induced or repressed by hypoxia, differential display polymerase chain reaction (DD-PCR) was used with mRNAs from the marine mussel, Mytilus galloprovincialis, under oxygen depletion and normal oxygen conditions. In total, 107 cDNA clones were differentially expressed under hypoxic conditions relative to the control mussel group. The differentially expressed genes were analyzed to determine the effects of hypoxia. They were classified into five functional categories: information storage and processing, cellular processes and signaling, metabolism, predicted general function only, and function unknown. The differentially expressed genes were predominantly associated with cellular processing and signaling, and they were particularly related to the signal transduction mechanism, posttranslational modification, and chaperone functions. The observed differences in the DD-PCR of 10 genes (encoding elongation factor 1 alpha, heat shock protein 90, calcium/calmodulin-dependent protein kinase II, GTPase-activating protein, 18S ribosomal RNA, cytochrome oxidase subunit 1, ATP synthase, chitinase, phosphoglycerate/bisphosphoglycerate mutase family protein, and the nicotinic acetylcholine receptor) were confirmed by quantitative RT-PCR and their transcriptional changes in the mussels exposed to hypoxic conditions for 24-72 h were investigated. These results identify biomarker genes for hypoxic stress and provide molecular-level information about the effects of oxygen depletion on marine bivalves.

Changes in gene expression profile of medaka with acute toxicity of Arochlor 1260, a polychlorinated biphenyl mixture.

Differential gene expression profiling was performed with a cDNA microarray in the liver tissue of the medaka fish, Oryzias latipes, after exposure to Arochlor 1260, a polychlorinated biphenyl (PCB) mixture, which is used as a coolant and insulating fluid for transformers and capacitors and is classified as a persistent organic pollutant. Twenty-six differentially expressed candidate genes were identified. The expression of 12 genes was up-regulated and that of 14 genes was down-regulated. These genes are associated with the cytoskeleton, development, endocrine/reproduction, immunity, metabolism, nucleic acid/protein binding, and signal transduction, or are uncategorized. The transcription of molecular biomarkers known to be involved in endocrine disruption (e.g., vitellogenins, choriogenins, and estrogen receptor alpha) was highly up-regulated. The same tendencies in gene expression changes were observed with real-time quantitative PCR (qRT-PCR) analysis, which was conducted to examine 12 selected candidate genes. These genes could be used as molecular biomarkers for biological responses to toxic chemicals, especially endocrine disrupting and carcinogenic chemical contamination in aquatic environments.

Expressions of Oxidative Stress-Related Genes and Antioxidant Enzyme Activities in Mytilus Galloprovincialis (Bivalvia, Mollusca) Exposed to Hypoxia

BackgroundIn this study, we investigated transcription and enzyme level responses of mussels Mytilus galloprovincialis exposed to hypoxic conditions. Genes for catalase (CAT), cytochrome P450, glutathione S-transferase (GST), metallothionein, superoxide dismutase (SOD), cytochrome c oxidase subunit 1 (COX-1), and NADH dehydrogenase subunit 2 were selected for study. Transcriptional changes were investigated in mussels exposed to hypoxia for 24 and 48 h and were compared to changes in control mussels maintained at normal oxygen levels. Activities of CAT, GST, and SOD enzymes, and lipid peroxidation (LPO) were also investigated in mussels following exposure to hypoxia for 24, 48, and 72 h.ResultsRelative to the control group, the CAT activity decreased in all hypoxia treatments, while the activity of GST significantly increased in mussels exposed to hypoxia for 24 and 48 h, but decreased in those exposed for 72 h. The LPO levels were significantly higher in mussels in the 24- and 48-h hypoxia treatments than those in the control mussels, but there was no significant change in the SOD activities among all hypoxia treatments. Messenger RNA levels for the CAT, cytochrome P450, GST, metallothionein, and SOD genes were not significantly affected by hypoxic conditions for 48 h, but the expressions of the COX-1 and NADH dehydrogenase subunit 2 genes were significantly repressed in mussels in both the 24- and 48-h exposure treatments.ConclusionsThese results demonstrate the transcriptional stability and changes among several genes related to oxidative stress under oxygen-depletion conditions in M. galloprovincialis and provide useful information about the modulation of antioxidant enzyme activities induced by hypoxia in a marine animal.

Alveopora japonica beds thriving under kelp

Alveopora japonica Eguchi, 1968 (Scleractinia, Acroporidae) occurs in shallow benthic communities from Southern Taiwan to high latitude areas of Japan where it is usually rare and nested among algae and soft corals (Dai and Horng 2009). It is also a peculiar species among the scleractinians due to its association with Symbiodinium clades C, E, and F (RodriguezLanetty et al. 2003; Jeong et al. 2012). In October 2012, we found extensive A. japonica beds (Fig. 1a, b) at Biyangdo, located to the northwest of Jeju Island, South Korea (33 24¢5†N, 126 13¢8†E), where it formed a dense and almost monospecific carpet at 10–17 m in depth. These stands were punctuated by the regionally endemic kelp Ecklonia cava. Based on 63 quadrats (0.25 m), A. japonica covered 67 ± 4 % of the benthos over an area of >1 ha. Most of the colonies were <10 cm in diameter. Many recruits occurred where coral cover was low such as around the base of holdfasts of the kelp, which represented 4 % of the benthic community. Other major benthic categories included calcareous crustose coralline algae (18 %) and sand (8 %). Ecklonia cava is usually the dominant benthic species around Jeju Island. Recently, traditional fisherwomen, ‘Haenyeo,’ as well as local divers, have reported orally a gradual decline in the population of this economically and ecologically important kelp to the benefit of A. japonica. Apart from being one of the first reports on high dominance of scleractinian coral species at this latitude, this observation might also indicate a persistent shift from kelp forests to coral dominance, and the potential loss of an economically important endemic taxa.

Toxaphene affects the levels of mRNA transcripts that encode antioxidant enzymes in Hydra

We evaluated toxaphene-induced acute toxicity in Hydra magnipapillata. The median lethal concentrations of the animals (LC(50)) were determined to be 34.5 mg/L, 25.0 mg/L and 12.0 mg/L after exposure to toxaphene for 24 h, 48 h and 72 h, respectively. Morphological responses of hydra polyps to a range of toxaphene concentrations suggested that toxaphene negatively affects the nervous system of H. magnipapillata. We used real-time quantitative PCR of RNA extracted from polyps exposed to two concentrations of toxaphene (0.3 mg/L and 3 mg/L) for 24 h to evaluate the differential regulation of levels of transcripts that encode six antioxidant enzymes (CAT, G6PD, GPx, GR, GST and SOD), two proteins involved in detoxification and molecular stress responses (CYP1A and UB), and two proteins involved in neurotransmission and nerve cell differentiation (AChE and Hym-355). Of the genes involved in antioxidant responses, the most striking changes were observed for transcripts that encode GPx, G6PD, SOD, CAT and GST, with no evident change in levels of transcripts encoding GR. Levels of UB and CYP1A transcripts increased in a dose-dependent manner following exposure to toxaphene. Given that toxaphene-induced neurotoxicity was not reflected in the level of AChE transcripts and only slight accumulation of Hym-355 transcript was observed only at the higher of the two doses of toxaphene tested, there remains a need to identify transcriptional biomarkers for toxaphene-mediated neurotoxicity in H. magnipapillata. Transcripts that respond to toxaphene exposure could be valuable biomarkers for stress levels in H. magnipapillata and may be useful for monitoring the pollution of aquatic environments.

Identification of differentially expressed genes in liver of marine medaka fish exposed to benzo[a]pyrene

The construction of a subtractive cDNA library of organisms and the analysis thereof have proven meaningful in the search for clones whose expressions are regulated by exterior changes. The principal objective of this study was to identify the differentially expressed genes of marine medaka fish (Oryzias javanicus) under benzo[a]pyrene exposure conditions. Medaka fish were exposed to 100 μg/L of benzo[a]pyrene for 24 hr and its hepatic RNA was extracted; RNA from the livers of non-exposed medaka fish was also extracted. Both RNAs were employed in the construction of the subtractive cDNA library and the nucleotide sequences of differentially expressed clones were analyzed. Twenty-eight genes of the total differentially expressed clones were considered significant; in particular, the transcription of the cytochrome P450 1A gene was upregulated in a dose-dependent manner under various benzo[a]pyrene concentration exposure conditions. The 28 genes were then divided into 4 categories: (1) information storage and processing, (2) cellular processes and signaling, (3) metabolism, and (4) poorly characterized. The data reported herein provide general information regarding the effects of benzo[a]pyrene contamination on marine organisms, and constitute a primary step in the development of novel biomarkers for marine environmental pollution.

Transcriptional response of marine medaka (Oryzias javanicus) on exposure to toxaphene

Differential gene expression profiles were established from the head and liver tissues of the marine medaka fish (Oryzias javanicus) after its exposure to toxaphene, a persistent organic pollutant and insecticide, using differential display polymerase chain reaction. Twenty-seven differentially expressed genes were identified, which were associated with the cytoskeleton, development, metabolism, nucleic acid/protein binding, and signal transduction. Among these genes, those encoding molecular biomarkers known to be involved in metabolism, ATP hydrolysis, and protein regulation were strongly induced at the transcription level, and genes encoding one structural protein subunit or involved in lipid metabolism were strongly downregulated. The same trends in gene expression changes were observed with real-time PCR analysis of 12 selected clones. The genes identified could be used as molecular biomarkers of biological responses to polychlorinated camphene contamination in aquatic environments.

Oxidative stress and gene expression in diverse tissues of Oryzias javanicus exposed to 17β-estradiol

Transcriptional changes in the expression of stress-related genes (catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, metallothionein, and ubiquitin) in intestinal, liver and muscle tissues of Javanese medaka (Oryzias javanicus) exposed to 17-estradiol (E2) were investigated using real-time quantitative PCR. The lipid peroxidation levels and superoxide dismutase activity in liver tissues was also examined at various exposure concentrations. In fish exposed to 10, 100 and 1000 g/L E2 the level of catalase mRNA increased significantly in intestinal and muscle tissues, while expression of the glutathione peroxidase gene showed the opposite trend in liver and muscle tissues. Expression of the glutathione S-transferase gene increased in a dose-dependent manner in liver and muscle tissues of fish exposed to E2, relative to the non-exposed control group. The level of glutathione reductase mRNA increased markedly in the liver tissues of all fish exposed to 100 and 1000 g/L E2, but decreased in intestinal tissues. The metallothionein gene was strongly downregulated in intestinal and liver tissues of fish exposed to E2, but slightly upregulated in muscle tissues. In all fish exposed to 100 and 1000 g/L E2 expression of the ubiquitin gene increased markedly in liver and muscle tissue, but not in intestinal tissues. Lipid peroxidation and superoxide dismutase activities increased significantly in all 17-estradiol treatment groups, and were correlated to the exposure concentration. Thus, E2 exposure differentially affected the transcription of a range of stress-related genes in various tissues of Javanese medaka, suggesting that analysis of transcriptional changes in these genes could be used as a rapid assay of the effects of E2 exposure.