Validation of reference genes for quantitative real-time PCR in chemical exposed and at different age's brackish water flea Diaphanosoma celebensis.

Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), a primary approach for evaluating gene expression, requires an appropriate normalization strategy to confirm relative gene expression levels by comparison, and rule out variations that might occur in analytical procedures. The best option is to use a reference gene whose expression level is stable across various experimental conditions to compare the mRNA levels of a target gene. However, there is limited information on how the reference gene is differentially expressed at different ages (growth) in small invertebrates with notable changes such as molting.

Acute toxicity of bisphenol A and its structural analogues and transcriptional modulation of the ecdysone-mediated pathway in the brackish water flea Diaphanosoma celebensis.

Bisphenol A (BPA) is a representative endocrine disrupting chemical (EDC) that has estrogenic effects in aquatic animals. In recent years, due to the continuing usage of BPA, its analogues have been developed as alternative substances to replace its use. The molting process is a pivotal point in the development and reproduction of crustaceans.

Evaluation of 4-nonylphenol and bisphenol A toxicity using multiple molecular biomarkers in the water flea Daphnia magna.

Alkylphenols are well-known endocrine disruptors and may cause developmental and reproductive disorders in aquatic organisms. Daphnia magna is commonly used in ecotoxicological studies as a promising model species to investigate the effects of endocrine distruptors. In the present study, transcriptional modulation of eleven potential molecular indicators related to detoxification, antioxidant, development, and cellular stress was analyzed in D.

De novo transcriptome assembly of brackish water flea based on short-term cadmium and benzo[]pyrene exposure experiments.

To develop a brackish water flea as a promising model for marine monitoring, were exposed to two pollutants, cadmium (Cd) and benzo[]pyrene (BaP), which have different chemical characteristics and distinct modes of metabolic action on aquatic animals. Twenty-four hours after exposure to Cd (2 mg/L) or BaP (25 μg/L), whole body transcriptomes were analyzed. In total, 99.

Novel approach for evaluating pharmaceuticals toxicity using Daphnia model: analysis of the mode of cytochrome P450-generated metabolite action after acetaminophen exposure.

Because of its widespread use, the pharmaceutical acetaminophen (APAP) is frequently detected in aquatic environments. APAP can have serious physiological effects, such as reduced reproduction, low growth rates, and abnormal behavior, in aquatic organisms. However, the methods available for evaluation of the aquatic toxicity of APAP are of limited usefulness.

Complete mitochondrial genome of the hydrothermal vent barnacle (Cirripedia, Thoracica).

Thoracican barnacles are common in hydrothermal vent fields. Here, we characterized the first mitogenome of a hydrothermal vent barnacle. The mitogenome of was 15,585 bp in length and had the typical pancrustacean gene arrangement.

Comparative Analysis of Transcriptional Profile Changes in Larval Zebrafish Exposed to Zinc Oxide Nanoparticles and Zinc Sulfate.

Many studies of the toxic effects of zinc oxide nanoparticles (ZnO NPs) in aquatic organisms have been performed because of increasing ZnO NP use. However, the toxicological pathways are not understood. In this study, ZnO NPs were found to be more toxic than ZnSO to zebrafish larvae, but ZnO NP toxicity did not involve transcript alterations.

Developmental Toxicity of Zinc Oxide Nanoparticles to Zebrafish (Danio rerio): A Transcriptomic Analysis.

Zinc oxide nanoparticles (ZnO NPs) are being utilized in an increasing number of fields and commercial applications. While their general toxicity and associated oxidative stress have been extensively studied, the toxicological pathways that they induce in developmental stages are still largely unknown. In this study, the developmental toxicity of ZnO NPs to embryonic/larval zebrafish was investigated.

Effects of chlorpyrifos on life cycle parameters, cytochrome P450S expression, and antioxidant systems in the monogonont rotifer Brachionus koreanus.

Chlorpyrifos is a widely used organophosphorus insecticide for controlling diverse insect pests of crops. In the monogonont rotifer Brachionus koreanus, population growth retardation with the inhibition of lifespan, fecundity, and individual body size of ovigerous females was shown over 10 d in response to chlorpyrifos exposure. At the molecular and biochemical levels, the rotifer B.

Structural basis for the ligand-binding specificity of fatty acid-binding proteins (pFABP4 and pFABP5) in gentoo penguin.

Fatty acid-binding proteins (FABPs) are involved in transporting hydrophobic fatty acids between various aqueous compartments of the cell by directly binding ligands inside their β-barrel cavities. Here, we report the crystal structures of ligand-unbound pFABP4, linoleate-bound pFABP4, and palmitate-bound pFABP5, obtained from gentoo penguin (Pygoscelis papua), at a resolution of 2.1 Å, 2.

Expression pattern of entire cytochrome P450 genes and response of defensomes in the benzo[a]pyrene-exposed monogonont rotifer Brachionus koreanus.

Cytochrome P450 (CYP) proteins are involved in the first line of detoxification mechanism against diverse polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene (B[a]P). In aquatic invertebrates, there is still a lack of knowledge on the CYP genes involved in the molecular response to B[a]P exposure due to limited gene information. In this study, we cloned the entire 25 CYP genes in the monogonont rotifer Brachionus koreanus with the aid of next generation sequencing (NGS) technologies and analyzed their transcript profiles with a real-time RT-PCR array to better understand B[a]P-triggered molecular response over different time courses.

Co-expression of antioxidant enzymes with expression of p53, DNA repair, and heat shock protein genes in the gamma ray-irradiated hermaphroditic fish Kryptolebias marmoratus larvae.

To investigate effects of gamma ray irradiation in the hermaphroditic fish, Kryptolebias marmoratus larvae, we checked expression of p53, DNA repair, and heat shock protein genes with several antioxidant enzyme activities by quantitative real-time RT-PCR and biochemical methods in response to different doses of gamma radiation. As a result, the level of gamma radiation-induced DNA damage was initiated after 4Gy of radiation, and biochemical and molecular damage became substantial from 8Gy. In particular, several DNA repair mechanism-related genes were significantly modulated in the 6Gy gamma radiation-exposed fish larvae, suggesting that upregulation of such DNA repair genes was closely associated with cell survival after gamma irradiation.

Evaluation of biomarker potential of cytochrome P450 1A (CYP1A) gene in the marine medaka, Oryzias melastigma exposed to water-accommodated fractions (WAFs) of Iranian crude oil.

CYP1A is involved in the metabolism of diverse chemicals, including polycyclic aromatic hydrocarbons and alkylated-PAHs, as a first line of detoxification mechanism. First, we identified and characterized the CYP1A gene from the marine medaka, Oryzias melastigma. O.

Genomic organization of selected genes in the small monogonont rotifer, Brachionus koreanus.

Information of genome structure with its size variation may provide important clues for evolutionary processes at lower taxon level in eukaryotes. Here, we analyzed the compact genome structure of the monogonont rotifer, Brachionus koreanus in the light of transphyletic genome comparison and economic genome usage. To confirm the genome compactness of B.

Susceptibility to oxidative stress and modulated expression of antioxidant genes in the copper-exposed polychaete Perinereis nuntia.

To identify and evaluate potentially useful biomarkers for oxidative stress as early warning indices in the polychaete, Perinereis nuntia, we exposed P. nuntia to copper (Cu) and measured several biomarker enzymes (glutathione S-transferase; GST, glutathione peroxidase; GPx, Metallothionein-like protein; MTLPs, and catalase; CAT) and genes (Pn-GSTs, Pn-CAT, and Pn-MT) with a cellular oxidative index, reactive oxygen species (ROS) level. Accumulated Cu concentrations in P.

The polychaete, Perinereis nuntia ESTs and its use to uncover potential biomarker genes for molecular ecotoxicological studies.

The polychaete, Perinereis nuntia, has been used as an indicator species to assess the environmental condition of benthic communities in coastal marine environments. Recently, high-throughput sequencing technology has been proven to be a useful method for analyzing expressed sequence tags (ESTs) in non-model species. Thus, we have obtained extensive cDNA information by the pyrosequencing method, to utilize the polychaete species as a test organism for sediment quality monitoring studies.

Analysis of expressed sequence tags from the liver and ovary of the euryhaline hermaphroditic fish, Kryptolebias marmoratus.

The self-fertilizing hermaphroditic fish, Kryptolebias marmoratus is considered a suitable model species in the fields of eco-biology, developmental biology, endocrinology, environmental genomics, aquatic toxicology, and molecular carcinogenesis. However, more extensive gene information is still needed to improve our understanding of the biology of this fish with respect to toxicological responses. We performed a transcriptomic study in this species using pyrosequencing.

Expression of superoxide dismutase (SOD) genes from the copper-exposed polychaete, Neanthes succinea.

To evaluate the superoxide dismutase (SOD) genes as a potential indicator of environmental stress, we cloned and sequenced the full-length cDNA of a Cu/Zn-SOD and Mn-SOD genes from the marine polychaete, Neanthes succinea. Ns-Cu/Zn- and Mn-SOD expression was measured using quantitative real-time RT-PCR in response to the oxidative stress induced by copper in the form of CuCl2. Ns-Cu/Zn-SOD expression increased in a dose dependent manner but the increase of Mn-SOD expression was evident only at a high concentration of copper.

Response of glutathione S-transferase (GST) genes to cadmium exposure in the marine pollution indicator worm, Perinereis nuntia.

Glutathione S-transferase (GST) is a phase II enzyme that functions as a detoxicant by catalyzing the conjugation of reduced glutathione with a variety of xenobiotics via cysteine thiol. Molecular genetic approaches using gene biomarkers show substantial relevance as sensitive biomarkers for the indication of pollution levels. In order to use GSTs as molecular biomarkers for marine pollution monitoring, we cloned and sequenced the full-length cDNA of seven GST genes from the marine polychaete Perinereis nuntia.

Molecular and biochemical modulation of heat shock protein 20 (Hsp20) gene by temperature stress and hydrogen peroxide (H₂O₂) in the monogonont rotifer, Brachionus sp.

The monogonont rotifer, Brachionus sp. has been regarded as a potential model for reproductive physiology, evolution, and environmental genomics. To uncover the role of the heat shock protein upon temperature stress and hydrogen peroxide (H₂O₂) exposure, we cloned heat shock protein 20 (Hsp20) and determined its modulatory response under different temperatures and H₂O₂ concentrations.

Ultraviolet B retards growth, induces oxidative stress, and modulates DNA repair-related gene and heat shock protein gene expression in the monogonont rotifer, Brachionus sp.

Ultraviolet B (UV-B) radiation causes direct cellular damage by breakage of DNA strands and oxidative stress induction in aquatic organisms. To understand the effect of UV-B radiation on the rotifer, Brachionus sp., several parameters including 24-h survival rate, population growth rate, and ROS level were measured after exposure to a wide range of UV-B doses.

Endocrine disrupting chemicals modulate expression of O⁶-methylguanine DNA methyltransferase (O⁶-MGMT) gene in the hermaphroditic fish, Kryptolebias marmoratus.

O⁶-methylguanine-DNA methyltransferase (O⁶-MGMT; EC 2.1.1.

Bisphenol A modulates expression of gonadotropin subunit genes in the hermaphroditic fish, Kryptolebias marmoratus.

Pituitary gonadotropins (GTHs), follicle stimulating hormone beta (FSH-beta), and luteinizing hormone beta (LH-beta) are the key hormones in the hypothalamus-pituitary-gonad (HPG) axis, and form the heterodimers between a common alpha subunit (gonadotropin-alpha) and FSH-beta and/or LH-beta. To obtain a better understanding on the modulation of gonadotropin subunit genes expression upon bisphenol A (BPA) exposure in hermaphroditic fish, we studied differential regulation of gonadotropin subunit genes from Kryptolebias marmoratus after the exposure of several EDCs. Expression profiles of these three genes when using quantitative real-time RT-PCR revealed that brain/pituitary tissues were highly expressed in these genes compared to other tissues.