Peering into molecular mechanisms of action with frogSCOPE

Helbing, Caren C.; Maher, Stacey K.; Han, Jun; Gunderson, Mark P.; Borchers, Christoph H.

doi:10.1016/j.ygcen.2010.01.012

Cited by 16 publications

(15 citation statements)

References 58 publications

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“…The confirmed stock concentrations were ATZ: 10,137 mg/L and 101 mg/L; DEA: 6,400 mg/L and 64 mg/L; and EE 2 : 3.29 mg/L and we will refer to the nominal concentrations in the manuscript. T 3 was prepared as previously described (Veldhoen et al, 2006) and the biological activity of this chemical was confirmed through measurement of TRβ transcript level induction as was previously described (Helbing et al, 2010). All exposures had the same volume of stock solution applied and contained 0.001% (vol/vol) IPA vehicle control (a 100,000X dilution).…”

Section: Methodsmentioning

confidence: 99%

Effect of low dose exposure to the herbicide atrazine and its metabolite on cytochrome P450 aromatase and steroidogenic factor-1 mRNA levels in the brain of premetamorphic bullfrog tadpoles (Rana catesbeiana)

et al. 2011

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The transcriptional regulator steroidogenic factor 1 (SF-1) and the enzyme cytochrome P450 aromatase (CYP19) play a central role in modulation of a broad range of tissue-specific developmental processes associated with hormone homeostasis that includes differentiation of the central nervous system. SF-1 and CYP19 expression may be targeted by a variety of endocrine disruptive agents prevalent within the environment. In the present study, we cloned and characterized partial sequences for bullfrog (Rana catesbeiana) SF-1 and CYP19 and examined the effects of a 48 h exposure to 1 and 100 μg/L of the herbicide atrazine (ATZ) and its major metabolite desethylatrazine (DEA), as well as 5 ng/L of the estrogenic chemical, 17α-ethynylestradiol (EE2), and 673 ng/L of the thyroid hormone, 3,5, 3′-triiodothyronine (T3), on SF-1 and CYP19 mRNA abundance in the brains of premetamorphic bullfrog tadpoles. Quantitative RT-PCR analysis showed an increase in CYP19 mRNA following a 48 h exposure to EE2 but not T3 while no significant changes in SF-1 transcript levels occurred. We observed a strong positive correlation between CYP19 and SF-1 transcript abundance in the ATZ-exposed animals which was not evident with DEA- or hormone-exposed tadpoles. Our results are intriguing in light of reported behavioral changes in ATZ-exposed frogs and suggest that further research is warranted to examine the relationship and role of CYP19 and SF-1 in amphibian brain development.

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Section: Methodsmentioning

confidence: 99%

Effect of low dose exposure to the herbicide atrazine and its metabolite on cytochrome P450 aromatase and steroidogenic factor-1 mRNA levels in the brain of premetamorphic bullfrog tadpoles (Rana catesbeiana)

et al. 2011

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“…To our knowledge, no comprehensive metabolomic study has yet been conducted on metamorphosis [9]. We applied a global, mass spectrometry (MS)-based metabolomics approach, using ultra-performance liquid chromatography (UPLC) coupled to a quadrupole time-of-flight (Q-TOF) mass spectrometer, to identify metabolites in serum samples from Rana catesbeiana (North American bullfrogs) at different postembryonic developmental stages: from tadpoles to froglets.…”

Section: Introductionmentioning

confidence: 99%

Metabolomic insights into system-wide coordination of vertebrate metamorphosis

et al. 2014

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BackgroundAfter completion of embryogenesis, many organisms experience an additional obligatory developmental transition to attain a substantially different juvenile or adult form. During anuran metamorphosis, the aquatic tadpole undergoes drastic morphological changes and remodelling of tissues and organs to become a froglet. Thyroid hormones are required to initiate the process, but the mechanism whereby the many requisite changes are coordinated between organs and tissues is poorly understood. Metabolites are often highly conserved biomolecules between species and are the closest reflection of phenotype. Due to the extensive distribution of blood throughout the organism, examination of the metabolites contained therein provides a system-wide overview of the coordinated changes experienced during metamorphosis. We performed an untargeted metabolomic analysis on serum samples from naturally-metamorphosing Rana catesbeiana from tadpoles to froglets using ultraperformance liquid chromatography coupled to a mass spectrometer. Total and aqueous metabolite extracts were obtained from each serum sample to select for nonpolar and polar metabolites, respectively, and selected metabolites were validated by running authentic compounds.ResultsThe majority of the detected metabolites (74%) showed statistically significant abundance changes (padj < 0.001) between metamorphic stages. We observed extensive remodelling of five core metabolic pathways: arginine and purine/pyrimidine, cysteine/methionine, sphingolipid, and eicosanoid metabolism and the urea cycle, and found evidence for a major role for lipids during this postembryonic process. Metabolites traditionally linked to human disease states were found to have biological linkages to the system-wide changes occuring during the events leading up to overt morphological change.ConclusionsTo our knowledge, this is the first wide-scale metabolomic study of vertebrate metamorphosis identifying fundamental pathways involved in the coordination of this important developmental process and paves the way for metabolomic studies on other metamorphic systems including fish and insects.

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“…systems toxicology approach). This overall framework in aquatic toxicology must include some knowledge of the complex relationship between gene expression, protein abundance, and metabolites (Martyniuk and Denslow, 2009; Helbing et al, 2010; Ekman et al, 2011, Garcia-Reyero and Perkins, 2011; Katsiadaki et al, 2010). Although there is much to be learned, there have been significant advances in moving towards these goals in the past few years in order to adapt Omics approaches to field based environmental studies.…”

Section: Future Of Quantitative Proteomics In Aquatic Toxicologymentioning

confidence: 99%

DIGE and iTRAQ as biomarker discovery tools in aquatic toxicology

Martyniuk

Alvarez

Denslow

2012

Ecotoxicology and Environmental Safety

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Molecular approaches in ecotoxicology have greatly enhanced mechanistic understanding of the impact of aquatic pollutants in organisms. These methods+- have included high throughput Omics technologies, including quantitative proteomics methods such as 2D differential in-gel electrophoresis (DIGE) and isobaric tagging for relative and absolute quantitation (iTRAQ). These methods are becoming more widely used in ecotoxicology studies to identify and characterize protein bioindicators of adverse effects. In teleost fish, iTRAQ has been used successfully in different fish species (e.g. fathead minnow, goldfish, largemouth bass) and tissues (e.g. hypothalamus and liver) to quantify relative protein abundance. Of interest for ecotoxicology is that many proteins commonly utilized as bioindicators of toxicity or stress are quantifiable using iTRAQ on a larger scale, providing a global baseline of biological effect from which to assess additional changes in the proteome. This review highlights the successes to date for high throughput quantitative proteomics using DIGE and iTRAQ in aquatic toxicology. Current challenges for the iTRAQ method for biomarker discovery in fish are the high cost and the lack of complete annotated genomes for teleosts. However, the use of protein homology from teleost fishes in protein databases and the introduction of hybrid LTQ-FT (Linear ion trap –Fourier transform) mass spectrometers with high resolution, increased sensitivity, and high mass accuracy are able to improve significantly the protein identification rates. Despite these challenges, initial studies utilizing iTRAQ for ecotoxicoproteomics have exceeded expectations and it is anticipated that the use of non-gel based quantitative proteomics will increase for protein biomarker discovery and for characterization of chemical mode of action.

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Peering into molecular mechanisms of action with frogSCOPE

Cited by 16 publications

References 58 publications

Effect of low dose exposure to the herbicide atrazine and its metabolite on cytochrome P450 aromatase and steroidogenic factor-1 mRNA levels in the brain of premetamorphic bullfrog tadpoles (Rana catesbeiana)

Effect of low dose exposure to the herbicide atrazine and its metabolite on cytochrome P450 aromatase and steroidogenic factor-1 mRNA levels in the brain of premetamorphic bullfrog tadpoles (Rana catesbeiana)

Metabolomic insights into system-wide coordination of vertebrate metamorphosis

DIGE and iTRAQ as biomarker discovery tools in aquatic toxicology

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