Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
Chromatin modification enzymes are important regulators of gene expression and some are evolutionarily conserved from yeast to human. Saccharomyces cerevisiae is a major model organism for genome-wide studies that aim at the identification of target genes under the control of conserved epigenetic regulators. Ume6 interacts with the upstream repressor site 1 (URS1) and represses transcription by recruiting both the conserved histone deacetylase Rpd3 (through the co-repressor Sin3) and the chromatin-remodeling factor Isw2. Cells lacking Ume6 are defective in growth, stress response, and meiotic development. RNA profiling studies and in vivo protein-DNA binding assays identified mRNAs or transcript isoforms that are directly repressed by Ume6 in mitosis. However, a comprehensive understanding of the transcriptional alterations, which underlie the complex ume6Δ mutant phenotype during fermentation, respiration, or sporulation, is lacking. We report the protein-coding transcriptome of a diploid MAT a/α wild-type and ume6/ume6 mutant strains cultured in rich media with glucose or acetate as a carbon source, or sporulation-inducing medium. We distinguished direct from indirect effects on mRNA levels by combining GeneChip data with URS1 motif predictions and published high-throughput in vivo Ume6-DNA binding data. To gain insight into the molecular interactions between successive waves of Ume6-dependent meiotic genes, we integrated expression data with information on protein networks. Our work identifies novel Ume6 repressed genes during growth and development and reveals a strong effect of the carbon source on the derepression pattern of transcripts in growing and developmentally arrested ume6/ume6 mutant cells. Since yeast is a useful model organism for chromatin-mediated effects on gene expression, our results provide a rich source for further genetic and molecular biological work on the regulation of cell growth and cell differentiation in eukaryotes.
ISSN: 1617-4615

hal-01151619
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01151619
DOI : 10.1007/s00438-015-1051-5
PUBMED : 25957495

Éditeur(s) : HAL CCSD Oxford University Press (OUP)
Résumé : International audience
The nature of factors governing the tempo and mode of protein evolution is a fundamental issue in evolutionary biology. Specifically, whether or not interactions between different sites, or epistasis, are important in directing the course of evolution became one of the central questions. Several recent reports have scrutinized patterns of long-term protein evolution claiming them to be compatible only with an epistatic fitness landscape. However, these claims have not yet been substantiated with a formal model of protein evolution. Here, we formulate a simple covarion-like model of protein evolution focusing on the rate at which the fitness impact of amino acids at a site changes with time. We then apply the model to the data on convergent and divergent protein evolution to test whether or not the incorporation of epistatic interactions is necessary to explain the data. We find that convergent evolution cannot be explained without the incorporation of epistasis and the rate at which an amino acid state switches from being acceptable at a site to being deleterious is faster than the rate of amino acid substitution. Specifically, for proteins that have persisted in modern prokaryotic organisms since the last universal common ancestor for one amino acid substitution approximately ten amino acid states switch from being accessible to being deleterious, or vice versa. Thus, molecular evolution can only be perceived in the context of rapid turnover of which amino acids are available for evolution.
ISSN: 0737-4038

hal-01544755
https://hal.archives-ouvertes.fr/hal-01544755
DOI : 10.1093/molbev/msu318

Éditeur(s) : HAL CCSD American Society for Biochemistry and Molecular Biology
Résumé : International audience
Polycyclic aromatic hydrocarbons (PAHs) such as benzo(a)pyrene (B(a)P) are widely distributed environmental contaminants, known as potent ligands of the aryl hydrocarbon receptor (AhR). These chemicals trigger an early and transient increase of intracellular calcium concentration ([Ca(2+)](i)), required for AhR-related effects of PAHs. The mechanisms involved in this calcium mobilization were investigated in the present study. We demonstrated that B(a)P-mediated [Ca(2+)](i) induction was prevented in endothelial HMEC-1 cells by counteracting β2-adrenoreceptor (β2ADR) activity using pharmacological antagonists, anti-β2ADR antibodies, or siRNA-mediated knockdown of β2ADR expression; by contrast, it was strongly potentiated by β2ADR overexpression in human kidney HEK293 cells. B(a)P was shown, moreover, to directly bind to β2ADR, as assessed by in vitro binding assays and molecular modeling. Pharmacological inhibition and/or siRNA-mediated silencing of various signaling actors acting downstream of β2ADR in a sequential manner, such as G protein, adenylyl cyclase, Epac-1 protein, and inositol 1,4,5-trisphosphate (IP(3))/IP(3) receptor, were next demonstrated to prevent B(a)P-induced calcium signal. Inhibition or knockdown of these signaling elements, as well as the use of chemical β-blockers, were finally shown to counteract B(a)P-mediated induction of cytochrome P-450 1B1, a prototypical AhR target gene. Taken together, our results show that B(a)P binds directly to β2ADR and consequently utilizes β2ADR machinery to mobilize [Ca(2+)](i), through activation of a G protein/adenylyl cyclase/cAMP/Epac-1/IP(3) pathway. This β2ADR-dependent signaling pathway activated by PAHs may likely be crucial for PAH-mediated up-regulation of AhR target genes, thus suggesting a contribution of β2ADR to the health-threatening effects of these environmental pollutants.
ISSN: 0021-9258

hal-00682497
https://hal.archives-ouvertes.fr/hal-00682497
DOI : 10.1074/jbc.M111.319970
PUBMED : 22167199

Éditeur(s) : HAL CCSD Nature Publishing Group
Résumé : International audience
Although TRAIL (tumor necrosis factor (TNF)-related apoptosis inducing ligand) is a well-known apoptosis inducer, we have previously demonstrated that acidic extracellular pH (pHe) switches TRAIL-induced apoptosis to regulated necrosis (or necroptosis) in human HT29 colon and HepG2 liver cancer cells. Here, we investigated the role of RIPK1 (receptor interacting protein kinase 1), RIPK3 and PARP-1 (poly (ADP-ribose) polymerase-1) in TRAIL-induced necroptosis in vitro and in concanavalin A (Con A)-induced murine hepatitis. Pretreatment of HT29 or HepG2 with pharmacological inhibitors of RIPK1 or PARP-1 (Nec-1 or PJ-34, respectively), or transient transfection with siRNAs against RIPK1 or RIPK3, inhibited both TRAIL-induced necroptosis and PARP-1-dependent intracellular ATP depletion demonstrating that RIPK1 and RIPK3 were involved upstream of PARP-1 activation and ATP depletion. In the mouse model of Con A-induced hepatitis, where death of mouse hepatocytes is dependent on TRAIL and NKT (Natural Killer T) cells, PARP-1 activity was positively correlated with liver injury and hepatitis was prevented both by Nec-1 or PJ-34. These data provide new insights into TRAIL-induced necroptosis with PARP-1 being active effector downstream of RIPK1/RIPK3 initiators and suggest that pharmacological inhibitors of RIPKs and PARP-1 could be new treatment options for immune-mediated hepatitis.
ISSN: 1350-9047

inserm-00871432
http://www.hal.inserm.fr/inserm-00871432
DOI : 10.1038/cdd.2012.90
PUBMED : 22814620

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
In this paper, we consider a direct protein transfer process between cells in co-culture. Assuming that cells continually encounter each other, and from some hypotheses on cell-to-cell rules of transfer, we derive discrete and continuous Boltzmann-like integrodifferential equations. The novelty of this model is to take into account multiple transfer rules. This new transfer model is used to fit the experimental data of cell-to-cell protein transfer in breast cancer.
ISSN: 0092-8240

Droits : info:eu-repo/semantics/OpenAccess
hal-01683780
https://hal.univ-antilles.fr/hal-01683780
https://hal.univ-antilles.fr/hal-01683780/document
https://hal.univ-antilles.fr/hal-01683780/file/MNPZF%20Bull%20Math%20Biol%202017.pdf
DOI : 10.1007/s11538-017-0319-9

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Among persisting questions on bone calcification, a major one is the link between protein expression and mineral deposition. A cell culture system is here proposed opening new integrative studies on biomineralization, improving our knowledge on the role played by non-collagenous proteins in bone. This experimental in vitro model consisted in human primary osteoblasts cultured for 60 days at the surface of a 3D collagen scaffold mimicking an osteoid matrix. Various techniques were used to analyze the results at the cellular and molecular level (adhesion and viability tests, histology and electron microscopy, RT-and qPCR) and to characterize the mineral phase (histological staining, EDX, ATG, SAED and RMN). On long term cultures human bone cells seeded on the osteoid-like matrix displayed a clear osteoblast phenotype as revealed by the osteoblast-like morphology, expression of specific protein such as alkaline phosphatase and expression of eight genes classically considered as osteoblast markers, including BGLAP, COL1A1, and BMP2. Von Kossa and alizarine red allowed us to identify divalent calcium ions at the surface of the matrix, EDX revealed the correct Ca/P ratio, and SAED showed the apatite crystal diffraction pattern. In addition RMN led to the conclusion that contaminant phases were absent and that the hydration state of the mineral was similar to fresh bone. A temporal correlation was established between quantified gene expression of DMP1 and IBSP, and the presence of hydroxyapatite, confirming the contribution of these proteins to the mineralization process. In parallel a difference was observed in the expression pattern of SPP1 and BGLAP, which questioned their attributed role in the literature. The present model opens new experimental possibilities to study spatio-temporal relations between bone cells, dense collagen scaffolds, NCPs and hydroxyapatite mineral deposition. It also emphasizes the importance of high collagen density environment in bone cell physiology.
ISSN: 1932-6203

hal-01142641
http://hal.upmc.fr/hal-01142641
http://hal.upmc.fr/hal-01142641/document
http://hal.upmc.fr/hal-01142641/file/Silvent_2013_Collagen.pdf
DOI : 10.1371/journal.pone.0057344

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
The mycotoxin alternariol (AOH), a frequent contaminant in fruit and grain, is known to induce cellular stress responses such as reactive oxygen production, DNA damage and cell cycle arrest. Cellular stress is often connected to autophagy, and we employed the RAW264.7 macrophage model to test the hypothesis that AOH induces autophagy. Indeed, AOH treatment led to a massive increase in acidic vacuoles often observed upon autophagy induction. Moreover, expression of the autophagy marker LC3 was markedly increased and there was a strong accumulation of LC3-positive puncta. Increased autophagic activity was verified biochemically by measuring the degradation rate of long-lived proteins. Furthermore, AOH induced expression of Sestrin2 and phosphorylation of AMPK as well as reduced phosphorylation of mTOR and S6 kinase, common mediators of signaling pathways involved in autophagy. Transmission electron microscopy analyzes of AOH treated cells not only clearly displayed structures associated with autophagy such as autophagosomes and autolysosomes, but also the appearance of lamellar bodies. Prolonged AOH treatment resulted in changed cell morphology from round into more star-shaped as well as increased β-galactosidase activity. This suggests that the cells eventually entered senescence. In conclusion, our data identify here AOH as an inducer of both autophagy and senescence. These effects are suggested to be to be linked to AOH-induced DSB (via a reported effect on topoisomerase activity), resulting in an activation of p53 and the Sestrin2-AMPK-mTOR-S6K signaling pathway.
ISSN: 0300-483X

hal-01117627
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01117627
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01117627/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01117627/file/Autophagy%20and%20senescence_accepted.pdf
PUBMED : 25456271

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 Elsevier
Résumé : International audience
All-trans retinoic acid (atRA) is the active form of vitamin A, known to activate retinoid receptors, especially the heterodimer retinoid X receptor (RXR):retinoic acid receptor (RAR) that otherwise may play a role in regulation of some drug transporters. The present study was designed to characterize the nature of human hepatic transporters that may be targeted by atRA and the heterodimer RXR:RAR. Exposure of human hepatoma HepaRG cells and primary human hepatocytes to 5μM atRA down-regulated mRNA levels of various sinusoidal solute carrier (SLC) influx transporters, including organic anion transporting polypeptide (OATP) 2B1, OATP1B1, organic cation transporter (OCT) 1 and organic anion transporter (OAT) 2, and induced those of the canalicular breast cancer resistance protein (BCRP). The retinoid concomitantly reduced protein expression of OATP2B1 and OATP1B1 and activity of OATPs and OCT1 and induced BCRP protein expression in HepaRG cells. Some transporters such as OATP1B3 and the bile salt export pump (BSEP) were however down-regulated by atRA in primary human hepatocytes, but induced in HepaRG cells, thus pointing out discrepancies between these two liver cell models in terms of detoxifying protein regulation. atRA-mediated repressions of OATP2B1, OATP1B1, OAT2 and OCT1 mRNA expression were finally shown to be counteracted by knocking-down expression of RARα and RXRα through siRNA transfection in HepaRG cells. atRA thus differentially regulated human hepatic drug transporters, mainly in a RXR:RAR-dependent manner, therefore establishing retinoids and retinoid receptors as modulators of liver drug transporter expression.
ISSN: 0928-0987

inserm-00804935
http://www.hal.inserm.fr/inserm-00804935
DOI : 10.1016/j.ejps.2013.01.005
PUBMED : 23352986

Éditeur(s) : HAL CCSD
Résumé : International audience
Despite their dynamic nature, certain chromatin marks must be maintained over the long term. This is particulary true for histone 3 lysine 9 (H3K9) trimethylation, that is involved in the maintenance of healthy differentiated cellular states by preventing inappropriate gene expression, and has been recently identified as the most efficient barrier to cellular reprogramming in nuclear transfer experiments. We propose that the capacity of the enzymes SUV39H1/2 to rebind to a minor fraction of their products, either directly or via HP1α/β, contributes to the solidity of this mark through (i) a positive feedback involved in its establishment by the mutual enforcement of H3K9me3 and SUV39H1/2 and then (ii) a negative feedback sufficient to strongly stabilize H3K9me3 heterochromatin in post-mitotic cells by generating local enzyme concentrations capable of counteracting transient bursts of demethylation. This model does not require direct molecular interactions with adjacent nucleosomes and is favoured by a series of additional mechanisms including (i) the protection of chromatin-bound SUV39H1/2 from the turnovers of soluble proteins, which can explain the uncoupling between the cellular contents in SUV39H1 mRNA and protein; (ii) the cooperative dependence on the local density of the H3K9me3 of HP1α/β-dependent heterochomatin condensation and, dispensably (iii) restricted enzyme exchanges with chromocenters confining the reactive bursts of SUV39H1/2 in heterochromatin. This mechanism illustrates how seemingly static epigenetic states can be firmly maintained by dynamic and reversible modifications. © 2016 Elsevier B.V.
Biochimica et Biophysica Acta - Gene Regulatory Mechanisms

hal-01467548
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01467548
DOI : 10.1016/j.bbagrm.2016.11.006
PUBMED : 27876670

Éditeur(s) : HAL CCSD Wiley-VCH Verlag
Résumé : International audience
Proteomic profiling plays a decisive role in the elucidation of molecular signatures representative of a specific clinical context. MuStem cell-based therapy represents a promising approach for clinical applications to cure Duchenne Muscular Dystrophy (DMD). To expand our previous studies collected in the clinically relevant DMD animal model, we decided to investigate the skeletal muscle proteome four months after systemic delivery of allogenic MuStem cells. Quantitative proteomics with isotope-coded protein labelling (ICPL) was used to compile quantitative changes in the protein expression profiles of muscle in transplanted Golden Retriever Muscular Dystrophy (GRMD) dogs as compared to GRMD dogs. A total of 492 proteins were quantified, including 25 that were overrepresented and 46 that were underrepresented after MuStem cell transplantation. Interestingly, this study demonstrates that somatic stem cell therapy impacts on the structural integrity of the muscle fascicle by acting on fibres and its connections with the extracellular matrix. We also show that cell infusion promotes protective mechanisms against oxidative stress and favours the initial phase of muscle repair. This study allows us to identify putative candidates for tissue markers that might be of great value in objectively exploring the clinical benefits resulting from our cell-based therapy for DMD.
ISSN: 1615-9853

Droits : info:eu-repo/semantics/OpenAccess
hal-01332030
https://hal.archives-ouvertes.fr/hal-01332030
https://hal.archives-ouvertes.fr/hal-01332030/document
https://hal.archives-ouvertes.fr/hal-01332030/file/Quantitative%20proteome%20profiling%20of%20dystrophic%20dog%20skeletal%20muscle_accepted.pdf
DOI : 10.1002/pmic.201600002

Éditeur(s) : HAL CCSD American Society for Biochemistry and Molecular Biology
Résumé : International audience
Background: We used evolutionary analysis of alkaline phosphatase (tissue nonspecific alkaline phosphatase, TNSALP) to predict missense mutations leading to hypophosphatasia. Results: We found 469 sensitive positions and validated 99% of the 204 known mutations. Conclusion: It is a more powerful method than in silico models to validate missense mutations in TNSALP. Significance: Such an approach should be widely used to support genetic diagnostics. ALPL encodes the tissue nonspecific alkaline phosphatase (TNSALP), which removes phosphate groups from various substrates. Its function is essential for bone and tooth mineralization. In humans, ALPL mutations lead to hypophosphatasia, a genetic disorder characterized by defective bone and/or tooth mineralization. To date, 275 ALPL mutations have been reported to cause hypophosphatasia, of which 204 were simple missense mutations. Molecular evolutionary analysis has proved to be an efficient method to highlight residues important for the protein function and to predict or validate sensitive positions for genetic disease. Here we analyzed 58 mammalian TNSALP to identify amino acids unchanged, or only substituted by residues sharing similar properties, through 220 millions years of mammalian evolution. We found 469 sensitive positions of the 524 residues of human TNSALP, which indicates a highly constrained protein. Any substitution occurring at one of these positions is predicted to lead to hypophosphatasia. We tested the 204 missense mutations resulting in hypophosphatasia against our predictive chart, and validated 99% of them. Most sensitive positions were located in functionally important regions of TNSALP (active site, homodimeric interface, crown domain, calcium site, ...). However, some important positions are located in regions, the structure and/or biological function of which are still unknown. Our chart of sensitive positions in human TNSALP (i) enables to validate or invalidate at low cost any ALPL mutation, which would be suspected to be responsible for hypophosphatasia, by contrast with time consuming and expensive functional tests, and (ii) displays higher predictive power than in silico models of prediction.
ISSN: 0021-9258

hal-01545479
https://hal.archives-ouvertes.fr/hal-01545479
DOI : 10.1074/jbc.M114.576843

Éditeur(s) : HAL CCSD Oxford University Press (OUP)
Résumé : International audience
Background: The objective of the study was to determine the incidence of transaminase elevation during dengue, and its predictive factors. Methods: In 2013, a longitudinal study was performed using data from all cases of dengue seen in Cayenne Hospital. Cox proportional modeling was used. Signs of major transaminase elevation were defined as an increase in aspartate amino transferase (AST) or alanine amino transferase (ALT) concentration over 10 times the normal value (10N). Results: There were 1574 patients and 13 249 person-days of follow-up. The incidence rate for signs of trans-aminase elevation (10N) was 0.55 per 100 person-days. Six patients had major transaminase elevation with AST.1000 units (0.43 per 1000 patient-days), and 73 patients (4.6%) developed transaminase elevation with AST .10N. The variables independently associated with major transaminase elevation were hyponatremia, low platelets, dehydration, hematocrit increase, food intolerance, positive nonstructural protein 1 (NS1), age over 15 years and the notion of paracetamol intake. Conclusions: Although very frequent, the incidence of major transaminase elevation was lower than reported elsewhere perhaps because of good access to care, or of the particular serotype causing this epidemic. The patients with transaminase elevation tended to be older, more severe and taking paracetamol.
ISSN: 0035-9203

inserm-01475594
http://www.hal.inserm.fr/inserm-01475594
http://www.hal.inserm.fr/inserm-01475594/document
http://www.hal.inserm.fr/inserm-01475594/file/2016%2C%20Djossou%20-%20Incidence%20and%20predictive%20factors%20of%20transaminase.pdf
DOI : 10.1093/trstmh/trv117

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
Résumé : International audience
Estrogen receptor alpha (ERα) has been recognized now for several decades as playing a key role in reproduction and exerting functions in numerous nonreproductive tissues. In this review, we attempt to summarize the in vitro studies that are the basis of our current understanding of the mechanisms of action of ERα as a nuclear receptor and the key roles played by its two activation functions (AFs) in its transcriptional activities. We then depict the consequences of the selective inactivation of these AFs in mouse models, focusing on the prominent roles played by ERα in the reproductive tract and in the vascular system. Evidence has accumulated over the two last decades that ERα is also associated with the plasma membrane and activates non-nuclear signaling from this site. These rapid/nongenomic/membrane-initiated steroid signals (MISS) have been characterized in a variety of cell lines, and in particular in endothelial cells. The development of selective pharmacological tools that specifically activate MISS and the generation of mice expressing an ERα protein impeded for membrane localization have begun to unravel the physiological role of MISS in vivo. Finally, we discuss novel perspectives for the design of tissue-selective ER modulators based on the integration of the physiological and pathophysiological roles of MISS actions of estrogens.
Physiological Reviews

hal-01533249
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01533249
DOI : 10.1152/physrev.00024.2016
PUBMED : 28539435

Éditeur(s) : HAL CCSD American Society for Biochemistry and Molecular Biology
Résumé : International audience
alpha- and beta-spectrins are components of molecular scaffolds located under the lipid bilayer and named membrane skeletons. Disruption of these scaffolds through mutations in spectrins demonstrated that they are involved in the membrane localization or the maintenance of proteins associated with them. The ubiquitous alphaII-spectrin chain bears in its central region a unique domain that is sensitive to several proteases such as calpains or caspases. The conservation of this region in vertebrates suggests that the proteolysis of alphaII-spectrin by these enzymes could be involved in important functions. To assess the role of alphaII-spectrin cleavage in vivo, we generated a murine model in which the exons encoding the region defining this cleavage sensitivity were disrupted by gene targeting. Surprisingly, homozygous mice expressing this mutant alphaII-spectrin appeared healthy, bred normally, and had no histological anomaly. Remarkably, the mutant alphaII-spectrin assembles correctly into the membrane skeleton, thus challenging the notion that this region is required for the stable biogenesis of the membrane skeleton in nonerythroid cells. Our finding also argues against a critical role of this particular alphaII-spectrin cleavage in either major cellular functions or in normal development.
ISSN: 0021-9258

inserm-00284621
http://www.hal.inserm.fr/inserm-00284621
http://www.hal.inserm.fr/inserm-00284621/document
http://www.hal.inserm.fr/inserm-00284621/file/Meary_-_JBC_2007.pdf
PUBMED : 17374614

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Adult fish exhibit a strong neurogenic capacity due to the persistence of radial glial progenitor cells. In zebrafish, radial glial cells display well-established markers such as the estrogen-synthesizing enzyme (AroB) and the brain lipid binding protein (Blbp), which is known to strongly bind omega-3 polyunsaturated fatty acids such as docosahexaenoic acid (DHA). While Blpb is mainly described in the telencephalon of adult zebrafish, its expression in the remaining regions of the brain is poorly documented. The present study was designed to further investigate Blbp expression in the brain, its co-expression with AroB, and its link with radial glial cells proliferation in zebrafish. We generated a complete and detailed mapping of Blbp expression in the whole brain and show its complete co-expression with AroB, except in some tectal and hypothalamic regions. By performing PCNA and Blbp immunohistochemistry on cyp19a1b-GFP (AroB-GFP) fish, we also demonstrated preferential Blbp expression in proliferative radial glial cells in almost all regions studied. To our knowledge, this is the first complete and detailed mapping of Blbp-expressing cells showing strong association between Blbp and radial glial cell proliferation in the adult brain of fish. Given that zebrafish is now recognized models for studying neurogenesis and brain repair, our data provide detailed characterization of Blbp in the entire brain and open up a broad field of research investigating the role of omega-3 polyunsaturated fatty acids in neural stem cell activity in fish
ISSN: 1567-133X

hal-01237088
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01237088
DOI : 10.1016/j.gep.2015.11.003
PUBMED : 26611722

Résumé : Shrimp culture in New Caledonia has developed according to the "intensified" semi-intensive model which uses ponds that stretch over several hectares. Local shrimp farms are for the most part installed in "tannes", barren, highly saline areas of the back mangrove forest. These areas generally have impermeable silty clay soils that have biogeochemical features particularly appropriate for benthic life and are often immersed by tides. During the six months of the grow-out phase, the "pond ecosystem" undergoes an increase in potential, progressing from an initial state of "natural lagoon" to a "grow-out pond" that is highly enriched with degrading organic matter....
La crevetticulture calédonienne s'est développée sur le modèle semi intensif « intensifié » qui se pratique en bassins de plusieurs hectares. Les fermes aquacoles locales sont pour la majorité implantées sur des « tannes », zones salées et nues d'arrière mangrove. Ces étendues présentent des sols généralement limono-argileux imperméables dont les caractéristiques bio géochimiques sont le plus souvent appropriées à la vie benthique car fréquemment recouvertes par la marée. Au cours des six mois d'élevage, on assiste à une montée en puissance de l' « écosystème-bassin » qui passe d'un état initial de « lagune naturelle » à celui final d'un « bac d'élevage très enrichi » en produits de dégradation de la matière organique. Le premier « fauteur de troubles » dans un bassin est l'aliment, ou plutôt les conditions imparfaites de l'alimentation qui contribuent fortement à la détérioration de la qualité de la colonne d'eau et du sol et conduisent à des accumulations. Avec la succession de cycles « forcés » par souci de rentabilité, le bassin peut devenir extrêmement consommateur d'oxygène et des zones de plus en plus vastes se recouvrent de vases noires susceptibles de libérer des composés réduits toxiques limitant l'espace vie de la crevette. La dégradation des fonds de bassin n'est cependant pas une fatalité. Les boues bien gérées sont un atout pour le fonctionnement harmonieux d'un tel agro-système et en particulier pour le bien être du cheptel. Le maintien d'une qualité favorable de sédiment pour la production de L.stylirostris passe par la mise en assec d'une durée minimale d'au moins deux semaines. Cette période inter élevage de réhabilitation « à l'air » sera d'autant plus courte et efficace que la zootechnie « en eau » aura été soignée. La Demande en Oxygène du Sédiment (DOS), les Matières Aisément oxydables (MAO), le Redox, l'abondance de la méiofaune, le rapport Protéines/Glucides de la Matière Organique peuvent renseigner les aquaculteurs sur la qualité de leur « foncier ». Ces paramètres prometteurs sont cependant difficiles à mettre en oeuvre au sein des entreprises. Ils sont abordés ici dans le cadre d'un rappel didactique de certaines bases biologiques essentielles pour tirer le meilleur parti de l'assec.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2007/rapport-4399.pdf
http://archimer.ifremer.fr/doc/00000/4399/

Éditeur(s) : AgroParisTech, ABIES
Résumé : Aquaculture is one of the main supply of protein in feed for mankind, notably in Asia. It increases and had exceeded the level of production of world fisheries, which is decreasing. According to sites and rearing methods, aquaculture contributes to add value but also in some cases to impact negatively the aquatic ecosystems. In the coastal area, a rich and productive zone, but also a place of conflicts, this activity presents a high potential for development if it takes into account the rules of sustainability. The conditions of this sustainability may be cleared with appropriate methods of foresight analysis applied for each development project. The analysis of the professional experience of D. Lacroix for 33 years in aquaculture is structured in three steps. Firstly, the work is to check the scientific value of the research conducted under his responsibility in various contexts and for different responsibilities. This study refers to the requirements of a PhD. In the second part of the work, the results of few major projects (French West Indies, French Guyana….) are analysed with the support of the theory of translation (Callon et al) which allows to build a network of actors and to project it into the context. This way of study includes a special attention to the aspects of time planning, legitimacy and acceptability. Finally, the author examines how this knowledge gained from experience could help building a method for foresight analysis aiming at a shorter planning, a better timing and less means for future projects. To achieve this goal the author integrates the vision of the economist R. Passet which structures any project in an ecosystemic framework, including criteria of sustainability. The mix of these two approaches (Callon & Passet) entails in a new tool for foresight analysis, called Aster, which originality is to offer the possibility to test various scenarios and related consequences to help decision -makers. This new method of foresight analysis, is then tested on the well known case study of the development of prawn Macrobrachium rosenbergii in French Guyana. The test shows to be successful as it delivers precise and arguable recommendations about research and development for all stakeholders.
L'aquaculture est l'une des composantes principales de l'apport en protéines dans le monde, surtout en Asie. Elle croît, alors que la pêche, qu'elle a dépassée en tonnage comme en valeur, décline. Selon les écosystèmes et les méthodes employées, elle peut contribuer à valoriser, mais aussi à dégrader les milieux aquatiques. Dans la zone côtière, espace riche et productif au plan biologique mais convoité, l'aquaculture conserve un potentiel de développement si sa mise en œuvre intègre les principes de la durabilité. Celle-ci peut être éclairée par la prospective avec des méthodes d'analyse adaptées à chaque projet. L'analyse de l'expérience de 33 ans en aquaculture de D. Lacroix est faite en trois temps. D'abord, il s'agit de vérifier la qualité de ses travaux de recherche scientifique menés dans des contextes et avec des responsabilités variés en suivant la grille d'analyse de référence d'un doctorat. Ensuite, l'analyse consiste à prendre un recul critique sur les bilans de réalisations majeures (Antilles, Guyane…) en mobilisant l'approche de M. Callon dont la théorie de la traduction permet de construire le réseau des acteurs et de le projeter dans la problématique. Cette grille d'analyse est complétée par l'étude des aspects de temporalité, légitimité et d'acceptabilité pour chaque réalisation. Ce travail sur diverses situations d'intervention permet de préparer une réflexion sur la proposition méthodologique qui suit et son adaptation à des conditions variables. Enfin, les connaissances acquises sont synthétisées afin de construire une méthode de prospective qui permettrait de gagner en temps, en efficacité et en moyens sur des projets à venir. Pour cela, l'analyse s'appuie sur la vision de R. Passet, qui structure les éléments de tout projet dans la hiérarchie des composantes de la durabilité. Le système dynamique qui en résulte peut être alors croisé avec divers scénarios afin d'en simuler les conséquences et donc d‟éclairer les choix. Cette méthode, dite Aster, est testée sur le cas « modèle » et bien documenté de l'élevage de la crevette Macrobrachium rosenbergii en Guyane française. Elle permet d'aboutir à des recommandations précises et argumentées en matière de recherche comme de développement pour tout décideur impliqué.

Droits : 2010 The author, AgroParisTech, ABIES
http://archimer.ifremer.fr/doc/00032/14349/11633.pdf
http://archimer.ifremer.fr/doc/00032/14349/

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Background & Aims: The severity of liver diseases is exacerbated by the death of hepatocytes, which can be induced by the sensing of pathogen associated molecular patterns (PAMPs) derived from the gut microbiota. The molecular mechanisms regulating these cell death pathways are poorly documented. In this study, we investigated the role of the receptor interacting protein kinase 1 (RIPK1), a protein known to regulate cell fate decisions, in the death of hepatocytes using two in vivo models of PAMP-induced hepatitis. Methods: Hepatitis was induced in mice by independent injections of two different bacterial PAMPs: lipopolysaccharide (LPS) and unmethylated CpG oligodeoxynucleotide (CpG-DNA) motifs. The role of RIPK1 was evaluated by using mice specifically lacking RIPK1 in liver parenchymal cells (Ripk1(LPC-KO)). Administration of liposome-encapsulated clodronate served to investigate the role of Kupffer cells in the establishment of the disease. Etanercept, a tumor necrosis factor (TNF)-decoy receptor, was used to study the contribution of TNF-alpha during LPS-mediated liver injury. Results: Whereas RIPK1 deficiency in liver parenchymal cells did not trigger basal hepatolysis, it greatly sensitized hepatocytes to apoptosis and liver damage following a single injection of LPS or CpG-DNA. Importantly, hepatocyte death was prevented by previous macrophage depletion or by TNF inhibition. Conclusions: Our data highlight the pivotal function of RIPK1 in maintaining liver homeostasis in conditions of macrophage-induced TNF burst in response to PAMPs sensing. Lay summary: Excessive death of hepatocytes is a characteristic of liver injury. A new programmed cell death pathway has been described involving upstream death ligands such as TNF and downstream kinases such as RIPK1. Here, we show that in the presence of LPS liver induced hepatic injury was due to secretion of TNF by liver macrophages, and that RIPK1 acts as a powerful protector of hepatocyte death. This newly identified pathway in the liver may be helpful in the management of patients to predict their risk of developing acute liver failure. (C) 2017 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
ISSN: 0168-8278

hal-01532060
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01532060
DOI : 10.1016/j.jhep.2017.01.005
PUBMED : 28088582