Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Zebrafish, Danio rerio, is increasingly used as an animal model to study the effects of pharmaceuticals and environmental estrogens. As most of these estrogens have only been tested on human estrogen receptors (ERs), it is necessary to measure their effects on zebrafish ERs. In humans there are two distinct nuclear ERs (hERα and hERβ), whereas the zebrafish genome encodes three ERs, zfERα and two zfERβs (zfERβ1 and zfERβ2). In this study, we established HeLa-based reporter cell lines stably expressing each of the three zfERs. We first reported that estrogens more efficiently activate the zfERs at 28°C as compared to 37°C, thus reflecting the physiological temperature of zebrafish in wildlife. We then showed significant differences in the ability of agonist and antagonist estrogens to modulate activation of the three zfER isotypes in comparison to hERs. Environmental compounds (bisphenol A, alkylphenols, mycoestrogens) which are hER panagonists and hERβ selective agonists displayed greater potency for zfERα as compared to zfERβs. Among hERα selective synthetic agonists, PPT did not activate zfERα while 16α-LE2 was the most zfERα selective compound. Altogether, these results confirm that all hER ligands control in a similar manner the transcriptional activity of zfERs although significant differences in selectivity were observed among subtypes. The zfER subtype selective ligands that we identified thus represent new valuable tools to dissect the physiological roles of the different zfERs. Finally, our work also points out that care has to be taken in transposing the results obtained using the zebrafish as a model for human physiopathology.
Toxicology and Applied Pharmacology

hal-01063914
https://hal.archives-ouvertes.fr/hal-01063914
DOI : 10.1016/j.taap.2014.07.020

Éditeur(s) : HAL CCSD
Résumé : National audience
Congrès de la SPTC - Société de Pharmaco-Toxicologie Cellulaire
Paris, France

hal-01122677
https://hal.archives-ouvertes.fr/hal-01122677

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
ISSN: 0166-445X

hal-01679436
https://hal.univ-antilles.fr/hal-01679436
DOI : 10.1016/j.aquatox.2016.04.006

Éditeur(s) : HAL CCSD Oxford University Press
Résumé : International audience
Brain Aromatase, Estrogens, and Behavior

hal-01122072
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01122072

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
The tg(cyp19a1b-GFP) transgenic zebrafish expresses GFP (green fluorescent protein) under the control of the cyp19a1b gene, encoding brain aromatase. This gene has two major characteristics: (i) it is only expressed in radial glial progenitors in the brain of fish and (ii) it is exquisitely sensitive to estrogens. Based on these properties, we demonstrate that natural or synthetic hormones (alone or in binary mixture), including androgens or progestagens, and industrial chemicals induce a concentration-dependent GFP expression in radial glial progenitors. As GFP expression can be quantified by in vivo imaging, this model presents a very powerful tool to screen and characterize compounds potentially acting as estrogen mimics either directly or after metabolization by the zebrafish embryo. This study also shows that radial glial cells that act as stem cells are direct targets for a large panel of endocrine disruptors, calling for more attention regarding the impact of environmental estrogens and/or certain pharmaceuticals on brain development. Altogether these data identify this in vivo bioassay as an interesting alternative to detect estrogen mimics in hazard and risk assessment perspective.
ISSN: 1932-6203

hal-00877371
https://hal.archives-ouvertes.fr/hal-00877371
https://hal.archives-ouvertes.fr/hal-00877371/document
https://hal.archives-ouvertes.fr/hal-00877371/file/Screening_Estrogenic_Activities-accepted.pdf
DOI : 10.1371/journal.pone.0036069

Éditeur(s) : HAL CCSD
Résumé : International audience
The present study was conducted to assess the effects of Cd exposure on estrogen signaling in the zebrafish brain, as well as the potential protective role of Zn against Cd-induced toxicity. For this purpose, the effects on transcriptional activation of the estrogen receptors (ERs), aromatase B (Aro-B) protein expression and molecular expression of related genes were examined in vivo using wild-type and transgenic zebrafish embryos. For in vitro studies, an ER-negative glial cell line (U251MG) transfected with different zebrafish ER subtypes (ERα, ERβ1 and ERβ2) was also used. Embryos were exposed either to estradiol (E2), Cd, E2+Cd or E2+Cd+Zn for 72 h and cells were exposed to the same treatments for 30 h. Our results show that E2 treatment promoted the transcriptional activation of ERs and increased Aro-B expression, at both the protein and mRNA levels. Although exposure to Cd, does not affect the studied parameters when administered alone, it significantly abolished the E2-stimulated transcriptional response of the reporter gene for the three ER subtypes in U251-MG cells, and clearly inhibited the E2 induction of Aro-B in radial glial cells of zebrafish embryos. These inhibitory effects were accompanied by a significant downregulation of the expression of esr1, esr2a, esr2b and cyp19a1b genes compared to the E2-treated group used as a positive control. Zn administration during simultaneous exposure to E2 and Cd strongly stimulated zebrafish ERs transactivation and increased Aro-B protein expression, whereas mRNA levels of the three ERs as well as the cyp19a1b remained unchanged in comparison with Cd-treated embryos. In conclusion, our results clearly demonstrate that Cd acts as a potent anti-estrogen in vivo and in vitro, and that Cd-induced E2 antagonism can be reversed, at the protein level, by Zn supplement.
Journal of Applied Toxicology

Droits : info:eu-repo/semantics/OpenAccess
hal-01274217
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01274217
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01274217/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01274217/file/Inhibitory%20effect%20of%20Cadmium%20on%20estrogen_accepted.pdf
DOI : 10.1002/jat.3285

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
The release of polycyclic aromatic hydrocarbons (PAHs) into the environment has increased very substantially over the last decades. PAHs are hydrophobic molecules which can accumulate in high concentrations in sediments acting then as major secondary sources. Fish contamination can occur through contact or residence nearby sediments or though dietary exposure. In this study, we analyzed certain physiological traits in unexposed fish (F1) issued from parents (F0) exposed through diet to three PAH mixtures at similar and environmentally relevant concentrations but differing in their compositions. For each mixture, no morphological differences were observed between concentrations. An increase in locomotor activity was observed in larvae issued from fish exposed to the highest concentration of a pyrolytic (PY) mixture. On the contrary, a decrease in locomotor activity was observed in larvae issued from heavy oil mixture (HO). In the case of the third mixture, light oil (LO), a reduction of the diurnal activity was observed during the setup of larval activity. Behavioral disruptions persisted in F1-PY juveniles and in their offspring (F2). Endocrine disruption was analyzed using cyp19a1b:GFP transgenic line and revealed disruptions in PY and LO offspring. Since no PAH metabolites were dosed in larvae, these findings suggest possible underlying mechanisms such as altered parental signaling molecule and/or hormone transferred in the gametes, eventually leading to early imprinting. Taken together, these results indicate that physiological disruptions are observed in offspring of fish exposed to PAH mixtures through diet.
ISSN: 0944-1344

hal-01121865
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01121865
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01121865/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01121865/file/Exposures%20of%20zebrafish_accepted.pdf
DOI : 10.1007/s11356-015-4157-8

Éditeur(s) : HAL CCSD John Libbey Eurotext
Résumé : National audience
ISSN: 1635-0421

hal-01118597
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01118597
DOI : 10.1684/ers.2012.0541

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Meconium is the first fecal excretion of newborns. This complex accumulative matrix allows assessing the exposure of the fetus to xenobiotics during the last 6 months of pregnancy. To determine the eventual effect of fetal exposure to micropollutants in this matrix, robust and sensitive analytical methods must be developed. This article describes the method development of liquid chromatography methods coupled to triple quadrupole mass spectrometry for relevant pollutants. The 28 selected target compounds had different physico-chemical properties from very polar (glyphosate) to non-polar molecules (pyrethroids). Tests were performed with three different types of columns: reversed phase, ion exchange and HILIC. As a unique method could not be determined for the simultaneous analysis of all compounds, three columns were selected and suitable chromatographic methods were optimized. Similar results were noticed for the separation of the target compounds dissolved in either meconium extract or solvent for reversed phase and ion exchange columns. However, for HILIC, the matrix had a significant influence on the peak shape and robustness of the method. Finally, the analytical methods were applied to “real” meconium samples.
ISSN: 0039-9140

hal-01141722
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01141722
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01141722/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01141722/file/Development%20of%20liquid%20chromatography%20%281%29.pdf
PUBMED : 25863396

Éditeur(s) : HAL CCSD
47. European Congress of Toxicology (EUROTOX 2011)
Paris, France

ineris-00970856
https://hal-ineris.archives-ouvertes.fr/ineris-00970856
INERIS : EN-2011-261

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Predictive toxicology aims at developing methodologies to relate the results obtained from in vitro experiments to in vivo exposure. In the case of polycyclic aromatic hydrocarbons (PAHs), a substantial amount of knowledge on effects and modes of action has been recently obtained from in vitro studies of gene expression. In the current study, we built a physiologically based toxicokinetic (PBTK) model to relate in vivo and in vitro gene expression in case of exposure to benzo(a)pyrene (BaP), a referent PAH. This model was calibrated with two toxicokinetic datasets obtained on rats exposed either through intratracheal instillation or through intravenous administration and on an in vitro degradation study. A good agreement was obtained between the model's predictions and the concentrations measured in target organs, such as liver and lungs. Our model was able to relate correctly the gene expression for two genes targeted by PAHs, measured in vitro on primary human macrophages and in vivo in rat macrophages after exposure to BaP. Combining in vitro studies and PBTK modeling is promising for PAH risk assessment, especially for mixtures which are more efficiently studied in vitro than in vivo.
Toxicology Letters

hal-00873317
https://hal.archives-ouvertes.fr/hal-00873317
https://hal.archives-ouvertes.fr/hal-00873317/document
DOI : 10.1016/j.toxlet.2010.11.017
PUBMED : 21134426

Éditeur(s) : HAL CCSD
Résumé : Benzophenone derivatives (BPs) are used as UV filters in many different products. These lipophilic compounds were recently described as ubiquitous contaminant of different compartiments of aquatic systems, including fish. In this study, we evaluated estrogenic activity of ten BPs in a model fish species, the zebrafish (Danio rerio), by combining newly developed in vitro and in vivo models. In vitro, three specific zebrafish reporter gene models expressing zebrafish estrogen receptors (ZfERs) in zebrafish hepatic cell line (ZFL) were used.Estrogenic activity of BPs was quantify in vivo by using a newly developed transgenic zebrafish line that expresses green fluorescent protein (GFP) under the control of the cyp19a1b promoter. The cyp19a1b gene codes for the brain aromatase B that is under the control of estrogens. Vitellogenin (Vtg), a marker of estrogenic compound, was used to assess the potential estrogenic effect of certain BPs in male fish. Five of 10 BPs induced an estrogenic activity on the three models: BP2, THB, BP1, 4BP, while BP and 234 BP were non active. We noted diffrence in BPs ranking depending on the zfER subtypes suggesting ?/? receptor selectivity by certain BPs. By quantifying GFP expression in live zebrafish, we confirm the estrogenic activity of some BP, i.e., BP1, THB, 4BP and 44'BP, as shown by their ability to induce GFP in a concentration-dependent manner. However, the ranking of BPs as a function of their estrogenic activity assay is different between models. For instance, 4BP was the most estrogenic compound in vivo but not in vitro. Differences were also noticed regarding the estrogenic activity of BP2. In vitro, BP2 has strong activity but had no effect on GFP in cyp19a1b-GFP embryo. However, BP2 induced strongly Vtg in male. The lack of estrogenic activity of BP2 in embryo may be related to difference of biodisponibility and/or metabolisation of this compound between embryo and adult fish. Our approach allows to characterize the estrogenic potency of emerging pollutants, using innovative in vitro and in vivo assays within a unique model fish species. The differences observed between assays deserve further studies to characterize the behavior of compounds in the different biological models in term of biodisponibility and metabolism.
1. European Conference on the Replacement, Reduction and Refinement of Animal Experiments in Ecotoxicology
Dübendorf, Switzerland

ineris-00971019
https://hal-ineris.archives-ouvertes.fr/ineris-00971019
INERIS : EN-2012-243

Éditeur(s) : HAL CCSD Edimark Santé
Résumé : National audience
Correspondance en métabolisme, hormone, diabète et nutrition

hal-01122040
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01122040

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
Meconium is the earliest stool of newborns, and is a complex matrix that reflects the degree of exposure of the fetus to xenobiotics. To investigate fetal exposure to volatile organic compounds, an analytical method was developed to identify and quantify BTEX (benzene, toluene, ethylbenzene, and o,m,p-xylene) and two chlorinated solvents (trichloroethylene and tetrachloroethylene) in meconium. Headspace-solid-phase microextraction coupled with gas chromatography-mass spectrometry was selected because it is simple, sensitive, can be automated, and requires no extensive sample preparation. Several extraction variables were optimized (fiber type, incubation time, temperature of fiber, and use of salt). Because meconium is a complex matrix, quantification by SPME was considered carefully because of potential interference, for example competitive adsorption. Calibration in water was compared with calibration in meconium using external and internal methods (with isotope-labeled compounds). In meconium, limits of quantification were determined to be in the range 0.064-0.096 ng g(-1) for the investigated compounds. All target compounds were determined in "real-case" meconium samples.
ISSN: 1618-2642

hal-01064967
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01064967
DOI : 10.1007/s00216-014-7836-2
PUBMED : 24838489

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Recent research has demonstrated the importance of dermal exposure for some semivolatile organic compounds (SVOCs) present in the gas phase of indoor air. Though models for estimating dermal intake from gaseous SVOCs exist, their predictions can be subject to variations in input parameters, which can lead to large variation in exposure estimations. In this sensitivity analysis for a steady state model, we aimed to assess these variations and their determinants using probabilistic Monte Carlo sampling for 8 SVOCs from different chemical families: phthalates, bisphenols, polycyclic aromatic hydrocarbons (PAHs), organophosphorus (OPs), organochlorines (OCs), synthetic musks, polychlorinated biphenyls (PCBs) and polybromodiphenylethers (PBDEs). Indoor SVOC concentrations were found to be the most influential parameters. Both Henry's law constant (H) and octanol/water partition coefficient (Kow) uncertainty also had significant influence. While exposure media properties such as volume fraction of organic matter in the particle phase (fom-part), particle density (ρpart), concentration ([TSP]) and transport coefficient (ɣd) had a slight influence for some compounds, human parameters such as body weight (W), body surface area (A) and daily exposure (t) make a marginal or null contribution to the variance of dermal intake for a given age group. Inclusion of a parameter sensitivity analysis appears essential to reporting uncertainties in dermal exposure assessment.
ISSN: 0160-4120

hal-01635769
https://hal.archives-ouvertes.fr/hal-01635769
https://hal.archives-ouvertes.fr/hal-01635769/document
https://hal.archives-ouvertes.fr/hal-01635769/file/Pelletier%202017%20Dermal%20expo%20rev2.pdf
DOI : 10.1016/j.envint.2017.02.005

Éditeur(s) : HAL CCSD John Libbey Eurotext
Résumé : International audience
Environmental health risk assessment is commonly used in France for risk management purposes. The Methodology section of the "Société française de santé environnement" (French Society for Environmental Health) seeks to improve the utility and implementation of risk assessment in France. An initial operational objective is to make a list of short-, intermediate- and long-term improvements. This inventory is based in part on the experience and expertise of society members in this area and in part on their analysis of "Science and Decisions: Advancing Risk Assessment" (National Research Council 2009). In this paper, the Society recommends modifications in the indications for and use of risk assessments, during each of the four stages of the process: hazard identification and dose-response assessment, exposure assessment, risk characterization, and results interpretation. Some may be implemented now, while others need consensus building or research because of gaps in knowledge. Environmental health risk assessment must be understood as a decision tool for risk management in situations of uncertainly. Thus, appropriate linkage is required between decision making and assessment, and between the methods and the expected results.
L'évaluation des risques sanitaires est fréquemment utilisée en France dans les processus de décision en santé environnementale. La section " Méthodologie d'évaluation des risques sanitaires " de la Société française de santé environnement (SFSE) s'est donnée pour objectif de proposer des éléments de réflexion et des outils méthodologiques en vue d'améliorer l'utilité et les pratiques de l'évaluation des risques sanitaires. Un premier objectif opérationnel est l'inventaire des améliorations possibles à court, moyen et long termes à apporter à la démarche et à son utilisation. Ce travail s'est fondé, d'une part, sur la pratique et l'expertise des membres de la section, et, d'autre part, sur une lecture critique du rapport " Science and Decisions : Advancing Risk Assessment " publié en 2009 par le National Research Council et portant également sur les évolutions de la démarche. Dans cet article, la section restitue les conclusions de cet inventaire critique et propose un certain nombre d'améliorations en termes d'indications et d'utilisations de l'évaluation des risques sanitaires pour chacune des quatre étapes : identification des dangers et relation dose-réponse ; évaluation de l'exposition ; caractérisation des risques ; et interprétation des résultats. Certaines recommandations peuvent être mises en oeuvre immédiatement ; d'autres, en préalable à leur mise en oeuvre, nécessitent un consensus ou encore de la recherche car les connaissances manquent. En conclusion, la section insiste sur le fait que l'évaluation des risques sanitaires doit être comprise comme une aide à la décision en situation d'incertitude ; l'articulation entre le contexte décisionnel et l'évaluation, sa méthodologie et les résultats attendus est donc primordiale.
ISSN: 1635-0421

ineris-00963325
https://hal-ineris.archives-ouvertes.fr/ineris-00963325
DOI : 10.1684/ers.2011.0435
INERIS : EN-2012-016

Éditeur(s) : HAL CCSD John Libbey Eurotext
Résumé : International audience
Chemical risk assessment is a tool used in decision-making processes concerning situations of scientific uncertainty. Approaches to risk assessment in occupational and environmental health share the same objective of risk prevention. Nevertheless, the methods for both differ depending on whether the goal is to enforce regulations or conduct expert assessments and studies. These differences are stronger in occupational health, for which regulations require risk assessments at the work station, to classify and prioritize exposure situations to implement occupational risk prevention strategies. Risk assessment thus has somewhat different meanings in occupational and environmental health. This article attempts to clarify the different terms used in the different contexts and to describe the similarities and differences of the two approaches.
ISSN: 1635-0421

hal-00936256
https://hal-univ-rennes1.archives-ouvertes.fr/hal-00936256
DOI : 10.1684/ers.2013.0646

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Endocrine disrupting chemicals and especially oestrogen receptor (ER) agonists have been extensively studied over the years due to their potential effects on sexual development and reproduction in vertebrates, notably fish. As ER agonists can exist as complex mixtures in the aquatic environment, evaluating the impact of combined exposure on oestrogenic effects has become increasingly important. Use of predictive models such as concentration addition (CA) and independent action (IA) has allowed assessment of combined estrogenic effects of complex multi-compound mixtures of ER agonists in various fish in vitro and in vivo experimental models. The present work makes use of a transgenic zebrafish strain, tg(cyp19a1b-GFP), which expresses the green fluorescent protein (GFP) under the control of the cyp19a1b (brain aromatase or aromatase B) gene to determine the oestrogenic potency of ER agonists alone or in mixtures. In these studies, tg(cyp19a1b-GFP) zebrafish embryos were exposed for four days (from one to five days post fertilization) to five different oestrogenic chemicals; 17α-ethinylestradiol (EE2), 17β-estradiol (E2), estrone (E1), bisphenol A (BPA) and 4-tert-octylphenol (OP), and three mixtures of up to four of these compounds. The mixture of BPA, OP and E2 was also tested with primary cultures of rainbow trout hepatocytes by analysing the ER-mediated induction of the oestrogenic biomarker vitellogenin in order to compare the performance of the two methods for assessing oestrogenic effects of complex mixtures. The three tested mixtures were predominantly acting in an additive manner on the expression of GFP. Additivity was indicated by the overlap of the 95% confidence interval of the concentration response curves for the observed data with the CA and IA prediction models, and model deviation ratios within a factor of two for a majority of the mixture concentrations. However, minor deviations determined as more than additive effects for the mixture of EE2, E1 and E2 and less than additive effects for the mixture of BPA, OP, EE2 and E1 were observed at the higher mixture concentrations tested. The successful prediction of additivity by CA and IA in tg(cyp19a1b-GFP) zebrafish embryos and deviations at high mixture concentrations seemed to correspond well to results obtained in the rainbow trout hepatocyte assay. The present results clearly show the usefulness of combining predictive modelling and use of in vitro bioassays for rapid screening of oestrogenic effects of complex mixtures and environmental samples.
ISSN: 0166-445X

hal-00877351
https://hal.archives-ouvertes.fr/hal-00877351
DOI : 10.1016/j.aquatox.2013.05.001
PUBMED : 23721851

Éditeur(s) : HAL CCSD John Libbey Eurotext
Résumé : International audience
French toxicological reference values (VTR) and French occupational exposure limit values (VLEP) are indicators that are widely used in risk assessment for, respectively, human and occupational health. Neither value can replace the other. However, the information obtained for one can be very useful for the other. We propose to clarify the fundamental differences and similarities between these two values. To achieve this, we review first their history and then the methods used to derive them; we conclude by proposing some improvements to harmonize them. A case study of formaldehyde illustrates our analysis.
ISSN: 1635-0421

hal-00936267
https://hal-univ-rennes1.archives-ouvertes.fr/hal-00936267
DOI : 10.1684/ers.2013.0647

Éditeur(s) : HAL CCSD
Résumé : We present an experimental study of relations between hydraulic properties of clayey rocks and their deformation at bulk and micro scale. The experimentation is based on triaxial deformation of kaolinitic core samples, on permeability measurements, and on several methods of analysis of the microstructure. To produce the samples, remoulded kaolinite material is consolidated by axial loading up to 20 MPa in an oedometric cell. Then, in a triaxial cell, an undrained stress path is applied in order to induce shearing damage in the sample. The permeability of the sample is measured by fluid flow across the sample. Before and after these hydro-mechanical tests, the evolution and the homogeneity of the pore space and moreover the texture of the material are analysed by X-ray tomography, environmental scanning electron microscopy and thermic neutron diffraction. Contrary to expectations, the permeability at the sample scale does not vary significantly, although quite large deformation is reached. The final deformation is heterogeneous, with well-developed shear bands. It appears that the development of compacting and dilating zones generates a heterogeneous permeability field, which results in a negligible variation of the bulk permeability at the sample scale.
International Journal of Rock Mechanics and Mining Science

hal-00496407
https://hal.archives-ouvertes.fr/hal-00496407
DOI : 10.1016/j.ijrmms.2010.04.007