Determination of mRNA expression of DMRT93B, vitellogenin, and cuticle 12 in Daphnia magna and their biomarker potential for endocrine disruption

Kim, Jungkon; Kim, Young‐Hee; Lee, Sang‐Woo; Kwak, Kyunghee; Chung, Wook-Jin; Choi, Kyungho

doi:10.1007/s10646-011-0707-0

Cited by 34 publications

(29 citation statements)

References 38 publications

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“…Thus, Cutie is likely to be highly expressed in animals that have a higher demand but lower availability of Ca 2+ . Our result are consistent with previous studies that demonstrated differential expression of genes involved in molting, such as cuticle proteins and proteolytic enzymes, under other environmental stressors such as fungicides and metals (Soetaert et al 2007;Vandenbrouck et al 2009;Kim et al 2011). …”

Section: Cuticle Proteins: Cutie and Icp2supporting

confidence: 93%

Synergistic interactions of biotic and abiotic environmental stressors on gene expression

Altshuler

McLeod

Colbourne

et al. 2015

Genome

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Understanding the response of organisms to multiple stressors is critical for predicting if populations can adapt to rapid environmental change. Natural and anthropogenic stressors often interact, complicating general predictions. In this study, we examined the interactive and cumulative effects of two common environmental stressors, lowered calcium concentration, an anthropogenic stressor, and predator presence, a natural stressor, on the water flea Daphnia pulex. We analyzed expression changes of five genes involved in calcium homeostasis - cuticle proteins (Cutie, Icp2), calbindin (Calb), and calcium pump and channel (Serca and Ip3R) - using real-time quantitative PCR (RT-qPCR) in a full factorial experiment. We observed strong synergistic interactions between low calcium concentration and predator presence. While the Ip3R gene was not affected by the stressors, the other four genes were affected in their transcriptional levels by the combination of the stressors. Transcriptional patterns of genes that code for cuticle proteins (Cutie and Icp2) and a sarcoplasmic calcium pump (Serca) only responded to the combination of stressors, changing their relative expression levels in a synergistic response, while a calcium-binding protein (Calb) responded to low calcium stress and the combination of both stressors. The expression pattern of these genes (Cutie, Icp2, and Serca) were nonlinear, yet they were dose dependent across the calcium gradient. Multiple stressors can have complex, often unexpected effects on ecosystems. This study demonstrates that the dominant interaction for the set of tested genes appears to be synergism. We argue that gene expression patterns can be used to understand and predict the type of interaction expected when organisms are exposed simultaneously to natural and anthropogenic stressors.

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Section: Cuticle Proteins: Cutie and Icp2supporting

confidence: 93%

Synergistic interactions of biotic and abiotic environmental stressors on gene expression

Altshuler

McLeod

Colbourne

et al. 2015

Genome

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“…These two proteins are functionally related to reproduction and development [25]. Both miconazole and promethazine are known to target the calcium-binding CaM [17], [19], a highly conserved protein that in D .…”

Section: Discussionmentioning

confidence: 99%

“…Biosystems, Foster, CA, USA) in a 20 µL reaction volume according to the protocol recommended by the manufacturer. The primers for a 77 bp fragment of vitellogenin (GenBank accession number: DY037239; forward: 5′-GCGGACGAGGTTGCAAAG-3′ and reverse: 5′-AGGAGCAGGAAGATGTCGTTCT-3′), a 199 bp fragment for cuticle protein 12 (DW985490; forward: 5′-AGCCAGTGGAACTACG-3′ and reverse: 5′-TCCAGCATCATCAGCG-3′), and a 71 bp fragment for β-actin (AJ292554; forward: 5′-CCACACTGTCCCCATTTATGAAG-3′ and reverse: 5′-CGCGACCAGCCAAATCC-3′) were adopted from Soetaert et al [26] and Kim et al [25]. The reaction plate was subjected in StepOne (Applied BioSystems) real time PCR with reverse transcription.…”

Section: Methodsmentioning

confidence: 99%

“…Individual RNA and DNA contents were used as biochemical endpoints, as RNA content is a practical and relatively sensitive tool for assessing changes in bulk protein synthesis rate, metabolic performance and growth [24]. Gene expression of vitellogenin and cuticle protein were used as molecular endpoints since they are relevant as indicators of reproductive and developmental effects [25]. In Daphnia , expression of these genes has been used to diagnose endocrine disruption as a result of exposure to various contaminants, including pharmaceuticals [25], [26] and metals [27], but also as a part of phototoxicity and oxidative stress responses [25].…”

Section: Introductionmentioning

confidence: 99%

“…Gene expression of vitellogenin and cuticle protein were used as molecular endpoints since they are relevant as indicators of reproductive and developmental effects [25]. In Daphnia , expression of these genes has been used to diagnose endocrine disruption as a result of exposure to various contaminants, including pharmaceuticals [25], [26] and metals [27], but also as a part of phototoxicity and oxidative stress responses [25]. Alterations in gene expression can be a direct response or a downstream result of a toxic exposure [28] and could facilitate identification of toxic mechanisms and affected pathways within the adverse outcome pathway concept [29].…”

Section: Introductionmentioning

confidence: 99%

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Are Pharmaceuticals with Evolutionary Conserved Molecular Drug Targets More Potent to Cause Toxic Effects in Non-Target Organisms?

et al. 2014

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The ubiquitous use of pharmaceuticals has resulted in a continuous discharge into wastewater and pharmaceuticals and their metabolites are found in the environment. Due to their design towards specific drug targets, pharmaceuticals may be therapeutically active already at low environmental concentrations. Several human drug targets are evolutionary conserved in aquatic organisms, raising concerns about effects of these pharmaceuticals in non-target organisms. In this study, we hypothesized that the toxicity of a pharmaceutical towards a non-target invertebrate depends on the presence of the human drug target orthologs in this species. This was tested by assessing toxicity of pharmaceuticals with (miconazole and promethazine) and without (levonorgestrel) identified drug target orthologs in the cladoceran Daphnia magna. The toxicity was evaluated using general toxicity endpoints at individual (immobility, reproduction and development), biochemical (RNA and DNA content) and molecular (gene expression) levels. The results provide evidence for higher toxicity of miconazole and promethazine, i.e. the drugs with identified drug target orthologs. At the individual level, miconazole had the lowest effect concentrations for immobility and reproduction (0.3 and 0.022 mg L−1, respectively) followed by promethazine (1.6 and 0.18 mg L−1, respectively). At the biochemical level, individual RNA content was affected by miconazole and promethazine already at 0.0023 and 0.059 mg L−1, respectively. At the molecular level, gene expression for cuticle protein was significantly suppressed by exposure to both miconazole and promethazine; moreover, daphnids exposed to miconazole had significantly lower vitellogenin expression. Levonorgestrel did not have any effects on any endpoints in the concentrations tested. These results highlight the importance of considering drug target conservation in environmental risk assessments of pharmaceuticals.

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An in situ toxicity identification and evaluation water analysis system: Laboratory validation

Steigmeyer

Zhang

Daley³

et al. 2017

Enviro Toxic and Chemistry

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It is difficult to assess the toxicity of a single stressor and establish a strong stressor-causality link when multiple stressors coexist. Toxicity identification evaluation (TIE) methodology uses a series of chemical and physical manipulations to fractionate compounds within a matrix and systematically identify potential toxicants. The current US Environmental Protection Agency application of TIE can provide valuable information but often lacks ecological realism and is subject to laboratory-related artifacts. An in situ TIE device (iTIED) was designed to assess the sources of toxicity in aquatic ecosystems. For this laboratory validation, each unit was equipped with a sorbent resin chamber, an organism exposure chamber, a water collection container, and a peristaltic pump. Chemical analyses of water processed by each iTIED unit were compared with both lethal and sublethal molecular responses of the organisms. The compound removal effectiveness of different sorbent resins was also compared. In addition to successfully fractionating diverse chemical mixtures, the iTIED demonstrated a potential for early detection of molecular biomarkers, which could identify chronic toxicity that may go unnoticed in traditional TIE assays. Utilizing this novel in situ system will reduce the uncertainty associated with laboratory-based simulations and aid management efforts in targeting compounds that pose the greatest threat. Environ Toxicol Chem 2017;36:1636-1643. © 2016 SETAC.

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Determination of mRNA expression of DMRT93B, vitellogenin, and cuticle 12 in Daphnia magna and their biomarker potential for endocrine disruption

Cited by 34 publications

References 38 publications

Synergistic interactions of biotic and abiotic environmental stressors on gene expression

Synergistic interactions of biotic and abiotic environmental stressors on gene expression

Are Pharmaceuticals with Evolutionary Conserved Molecular Drug Targets More Potent to Cause Toxic Effects in Non-Target Organisms?

An in situ toxicity identification and evaluation water analysis system: Laboratory validation

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