Éditeur(s) : HAL CCSD
Résumé : Spermatogenesis in mammals is a complex biological function including cellular processes such as proliferation, meiosis and differentiation, aiming to the production of male gametes in the testis. If the seminiferous epithelium is well described in terms of organization and cellular morphology of cells that compose it, the processes by which undifferentiated diploid germ cells enter meiosis and give haploid cells that undergo many morphological transformations, are not fully decrypted. These processes rely on the coordinated and sequential expression of genes, including specific products for each stage of germ cell development These gene products are essential at each key stage of spermatogenesis. Transcriptomics since the 1990s, and proteomics since the 2000s have contributed to the improved. understanding of these mechanisms. A long term proteomic study aiming at characterizing the proteomes of Sertoli cells and germ cells, and a recent study that characterized and quantified the transcriptome of isolated rat testicular cells at high resolution using de novo sequencing of transcripts (RNA-Seq), have been the basis of my thesis work. The latter study showed the accumulation of long non-Coding RNAs (lncRNAs) and testicular unannotated transcripts (TUTs) at meiotic and post-Meiotic stages of spermatogenesis in the rat. In this context, my thesis work aimed at validating the coding potential of many genes expressed in germ cells using RNA-Seq combined with shotgun proteomics, a so-Called PIT (Proteomics Informed by transcriptomics) approach. In this approach, the protein sequences translated from the transcripts assembled by RNA-Seq in the different testicular cell types are integrated into a custom database of protein sequences used to query mass spectrometry data obtained from proteins of meiotic and post-Meiotic cells. The PIT approach showed that 69 TUTs or lncRNA (corresponding to 44 loci) code for proteins in meiotic cells and post meiotic cells, and we confirmed experimentally the meiotic and post-Meiotic expression for two new transcripts encoding for VAMP9, a protein of the SNARE family, and a new testicular enolase T-ENOL. The post-Meiotic expression of T-ENOL protein was confirmed by immunohistochemistry using a polyclonal antibody raised against the recombinant protein. This approach also allowed us to identify new isoforms of known proteins, specific to each stage of spermatogenesis. Germ cells and Sertoli cells maintain a dialogue which is necessary to the success of spermatogenesis and spermiogenesis. Another part of my work aimed at identifying membrane proteins, in germ cells and residual bodies, that may be involved in the dialogue between Sertoli cells and germ cells, using a ICPL relative quantification proteomic approach. The ICPL analysis enabled us to establish a list of 166 proteins whose expression is differential between pachytene spermatocytes, round spermatids and residual bodies. Their differential expression suggests that these proteins may play a role in spermiogenesis. Thanks to the Gene Ontology annotations, a list of 8 proteins with a putative role in signal transduction, cell recognition or differentiation, thus potentially involved in the dialogue between Sertoli and germ cells was drawn. In addition, I provided a first proteome of rat Sertoli cells, germ cells and residual bodies obtained by shotgun proteomics.
La spermatogenèse chez les mammifères est une fonction biologique complexe incluant des processus de prolifération cellulaire, de méiose et de différenciation uniques visant à la production des gamètes mâles au sein du testicule. Si l’épithélium séminifère est bien décrit sur le plan de son organisation et de la morphologie des cellules qui le composent, les processus par lesquels les cellules germinales diploïdes indifférenciées entrent en méiose pour donner ensuite des cellules haploïdes subissant par la suite de nombreuses transformations morphologiques, ne sont pas totalement décryptés. Ils reposent sur l’expression coordonnée et séquentielle de gènes dont les produits spécifiques de chaque stade de développement des cellules germinales sont essentiels aux étapes clés de la spermatogenèse. La transcriptomique depuis les années 1990 et la protéomique depuis les années 2000 ont contribué à l’amélioration de la connaissance de ces mécanismes. Une étude protéomique visant à caractériser par des approches systématiques et différentielles les protéomes des cellules de Sertoli et de la lignée germinale, et d’autre part une étude récente, réalisée dans notre unité, qui a permis de caractériser et de quantifier le transcriptome des cellules testiculaires isolées de rat en utilisant le séquençage de novo des transcrits (RNA-Seq), ont été à la base de mes travaux de thèse. Cette dernière étude a mis en évidence l’accumulation de longs ARNs non codants (lncRNAs) et de transcrits testiculaires non annotés (TUTs) aux stades méiotique et post- méiotique de la spermatogenèse chez le rat. Dans ce contexte, mon travail a consisté à valider le potentiel codant de nombreux gènes exprimés dans les cellules germinales par une approche dite PIT (Proteomics Informed by Transcriptomics) couplant protéomique Shotgun et RNA-Seq. Dans ce type d’approche, les séquences protéiques déduites des transcrits des différents types cellulaires, assemblés par RNA-Seq, sont intégrées dans une base personnalisée de séquences protéiques utilisée pour interroger les données de spectrométrie de masse obtenues à partir de protéines de cellules méiotiques et post-Méiotiques. L’approche PIT a permis de montrer que 69 TUTs ou lncRNA (correspondant à 44 loci) codent pour des protéines dans les cellules méiotiques et post méiotiques. L’expression post-Méiotique de deux nouveaux transcrits, l’un codant pour la protéine VAMP9, une protéine de la famille SNARE, et l’autre pour une nouvelle énolase T-ENOL a pu être confirmée. L’expression post-Méiotique de T-ENOL a été confirmée par immunohistochimie à l’aide d’un anticorps polyclonal produit contre la protéine recombinante. Cette approche nous a également permis d’identifier de nouvelles isoformes de protéines connues spécifiques de chaque stade de la spermatogenèse. Les cellules germinales et les cellules de Sertoli entretiennent le dialogue nécessaire au bon déroulement de la spermatogenèse. Une autre partie de mon travail a consisté à identifier des protéines membranaires des cellules germinales et des corps résiduels, susceptibles d’intervenir dans le dialogue entre les cellules de Sertoli et les cellules germinales, par une approche protéomique de quantification relative ICPL. Cette approche a permis d’établir une liste de 166 protéines différentiellement exprimées entre les spermatocytes pachytène, les spermatides rondes et les corps résiduels, qui sont susceptibles de jouer un rôle dans la spermiogénèse. Grâce aux annotations de le Gene Ontology, j’ai pu établir une liste de 8 protéines ayant un rôle supposé dans la transduction du signal, la reconnaissance cellulaire ou bien la différenciation. Par ailleurs, j’ai pu établir par protéomique Shotgun un premier protéome des cellules de Sertoli, des cellules germinales et des corps résiduels chez le rat.
https://tel.archives-ouvertes.fr/tel-01127389

Droits : info:eu-repo/semantics/OpenAccess
NNT : 2014REN1S064
tel-01127389
https://tel.archives-ouvertes.fr/tel-01127389
https://tel.archives-ouvertes.fr/tel-01127389/document
https://tel.archives-ouvertes.fr/tel-01127389/file/2014REN1S064.pdf

Éditeur(s) : HAL CCSD
Résumé : Due to their phylogenetic position outside from Bilateria, cnidarians and ctenophores are key phyla to understanding the origin and early evolution of molecular mechanisms, cell types, or body plan features at the level of Metazoa. Unlike other non-bilaterian phyla (sponges and placozoans), they possess neurons and muscle cells. The first part of my work relates to the problem of the origin of the metazoan nervous system. Recent studies have suggested a dual origin for nerve cells, with ctenophore neurons having emerged independently from those of cnidarians and bilaterians. This hypothesis relies on phylogenomic analyses supporting ctenophores as sister-group to all other metazoans, and on the purported absence in ctenophores of almost all well-known neuro-transmission systems, including the cholinergic system. Refuting this claim, I present substantial evidence in favour of acetylcholine (Ach) being used in the ctenophore Pleurobrachia pileus as an extra-cellular messenger, including within the nervous system. This conclusion arises from the conjunction of multiple approaches at the molecular scale (orthology analyses, Ach immuno-localisation, gene in situ hybridisation), at the cellular scale (calcium imaging on muscle cells exposed to Ach) and at the organism scale (pharmacological treatments with an Ach analogue and Ach antagonists). In the second part of my thesis, I address the conservation of molecular mechanisms involved in the control of cellular proliferation. Notably, the Hippo pathway is known for its role in bilaterians in the negative control of cellular proliferation. I show that components of this pathway are conserved in Pleurobrachia pileus and in Clytia hemisphaerica and that the nuclear vs. cytoplasmic localisation of the transcriptional co-factor Yorkie is correlated to the extent of proliferation within C. hemisphaerica tissues. Finally, RNAseq differential gene expression analyses along a cellular conveyor belt of the C. hemisphaerica medusa suggests a tendency for conservation of function among a set of genes whose mammalian orthologues have documented roles in the arrest of cellular proliferation.
Les Cnidaires et les Cténaires sont deux embranchements extérieurs aux Bilatériens qui, de par leur position phylogénétique, constituent des modèles d'étude cruciaux pour comprendre l'origine et l'évolution de mécanismes moléculaires ou encore de types cellulaires à grande échelle chez les Métazoaires. Ils sont les deux seuls embranchements non-Bilatériens à posséder des cellules nerveuses ainsi que des cellules musculaires. La première partie de mon travail de thèse s'inscrit dans le contexte de la compréhension de l'origine du système nerveux des Métazoaires. Des études récentes suggèrent une origine double du système nerveux ; ainsi celui des Cténaires aurait été acquis de manière convergente par rapport à celui des Cnidaires et des Bilatériens. Cela repose sur des données récentes de phylogénomique proposant les Cténaires en tant que groupe-frère de tous les autres Métazoaires et sur l'absence supposée chez les Cténaires de quasiment tous les systèmes de neurotransmission connus y compris le système cholinergique. A l'encontre de ces hypothèses, j'ai pu mettre en évidence l'existence d'un système cholinergique chez le Cténaire Pleurobrachia pileus ainsi que son étroite association avec le système nerveux. Cette conclusion repose sur une multiplicité d'approches à l'échelle moléculaire (analyses d'orthologie, immunolocalisation, hybridations in situ), cellulaire (imagerie calcique) et de l'organisme (approche pharmacologique). Dans la seconde partie de ma thèse, j'aborde la question de la conservation des mécanismes moléculaires impliqués dans le contrôle de la prolifération cellulaire à l'échelle des Métazoaires. J'ai pu mettre en évidence la conservation des acteurs.
https://tel.archives-ouvertes.fr/tel-01633223

NNT : 2015PA066629
tel-01633223
https://tel.archives-ouvertes.fr/tel-01633223
https://tel.archives-ouvertes.fr/tel-01633223v2/document
https://tel.archives-ouvertes.fr/tel-01633223/file/2015PA066629.pdf

Résumé : Bacteriological study of abyssal sediments: During the Demeraby mission of the N/O Jean Charcot in the western Atlantic Ocean, a study of the microflora of the digestive tracts of echinoderms (Holothuroidea of various genera, Asteroidea, Echinoidea) was compared with that of the surrounding sediment. The bacterial concentration observed in the digestive tracts of all the animals studied is higher than that in the surrounding sediments. The largest numbers were always found in the most anterior part of the digestive tracts of the holothurians. In some cases, the increase is considerable (7.10 versus 5.2. 10 cells per millilitre): It indicates a proliferation of bacterial cells in the anterior part of the digestive tract. As the sediment moves through the digestive tract, the bacterial level progressively decreases. In the most posterior part, the concentration is, however, always clearly greater than that in the surrounding sediment. In the case of the holothurian of the genus Deima, analysis according to the Adamsonian principle of comparison shows that the microflora isolated from the various parts of the digestive tract is composed of taxonomic groups showing little resemblance to those isolated from the sediment. In the case of the holothurian of the genus Pseudostichopus, no specialisation of the microflora from digestive tract contents appears with respect to that of the ingested sediment. Examination of metabolic activities suggests a juxtaposition of two types of communities: Free sediment - sediment in transit that complement one other in their enzymatic and nutritional behaviour Indeed, the strains of free sediment seem to be rather suitable for hydrolysing and using carbohydrates, while those of enteric communities prefer the hydrolysis of lipids and proteins and the assimilation of fatty acids, dicarboxylic acids, hydroxylic acids and certain amino acids. Moreover, the enteric microflorae possess a greater nutritional versatility that, linked with its total numbers, has a sizeable catabolic potential. .../... -2- Measurements of heterotrophic activity done by means of 14 C glucose tracers make it possible to show that the metabolic activities of the bacterial communities of superficial abyssal sediments vary according to zones. In the Amazon Cone, a clear glucose mineralization gradient over a distance of a few kilometres can be observed. In the abyssal plain of Demerara heterotrophic activities are slowed in relation to those of the superficial sediments of the Cone of the Amazon, even though the bacterial numbers and the depths are of the same order of magnitude in the two zones. The abundant microflorae associated with the various divisions of digestive tract contents of the Deima abyssal holothurian manifest little metabolic activity as regards the glucose. However, the bacterial communities of Pseudostichopus and Psychropotes manifest very strong heterotrophic activities, increasing from the oesophagus towards the cloaca. Current results are too fragmentary to allow us to observe possible physiological specialisation of enteric bacterial communities enabling the facilitation of digestion processes in the host. Conclusion: In the abyssal environment, characterised by poorly developed bacterial communities, microflorae associated with the digestive tract contents of echinoderms have considerably greater cellular concentrations. By means of their enzymatic equipment, these enteric communities, which complement those of the free microflorae of superficial sediments, can participate in the transformation of organic matter in the abyssal area. Study of the dynamics of microbial communities and their heterotrophic activities in shellfish farms: From experiments done on the water in the Arcachon basin, trials to quantify and describe the reactivity potential of bacterial communities were performed. Bacterial communities were defined by direct counts, counts after setting them up for cultivation, structure and diversity of these communities, dosage of AMP, ADP, ATP. Two opposite ecological conditions were considered: Autumn conditions (high temperature) and a winter conditions (low temperature). Tubs of sea water of 6 and 60 litres were enriched either with ammonium chloride or a mixture of amino acids, at about 500 uAtg N.L. The experiments lasted 24 hours, with sampling every 3 hours. The autumn community reacted rapidly (6 hours), the response to the enrichment in organic nitrogen being the highest. The structure of the community has unstable immature stages. On the contrary, during the winter experiment, no response could be perceived. This may mean that the latency phase is greater than 24 hours. The structure of this community has mature stages. In conclusion, bacterial communities react in the same way as macroscopic communities. Stable communities are diversified and correspond to mature stages of the ecosystem; the immature stages are unstable and less diversified. This type of experiment has made it possible to bring to light the rapidity of response of bacterial populations, and can be used to define the stability of a bacterial community in a precise biotope.
ETUDE BACTERIOLOGIQUE DES SEDIMENTS ABYSSAUX RESUME Au cours de la mission DEMERABY du n/o Jean CHARCOT en atlantique occidental , une étude de la microflore des tractus digestifs d'Echinodermes (Holothurides de différents genres, Astérides, Echinides) a été comparée à celle du sédiment environnant. La concentration bactérienne observée dans les tractus digestifs de tous les animaux étudiés est supérieure à celle des sédiments environnants. Les effectifs les plus importants sont toujours observés dans la partie la plus antérieure des tractus digestifs des holothuries. Dans certains cas, l'accroissement est considérable (7.10 contre 5,2. 10 cellules/ml) : il indique une prolifération des cellules bactériennes dans la partie antérieure du tractus digestif. Au.cours de la progression du sédiment dans le tractus digestif, la charge bactérienne décroit progressivement. Dans la partie la plus postérieure, la concentration est cependant toujours nettement supérieure à celle du sédiment environnant-. Dans le cas d'une holothurie du genre Deima, l'analyse selon le principe adansonien de comparaison montre que la microflore isolée des différentes parties du tractus digestif est constituée de groupes taxonomiques présentant peu de ressemblance avec ceux isolés du sédiment. Dans le cas d'une holothurie du genre Pseudostichopus il n'apparait pas de spécialisation de la microflore du contenu du tractus digestif par rapport à celle du sédiment ingéré. ¿ ¿ . L'étude des activités métaboliques suggère une juxtaposition de deux types de communautés : sédiment libre-sédiment en transit qui se complètent dans leur comportement enzymatique et nutritionnel. En effet les souches du sédiment libre semblent être plutôt aptes à hydrolyser et à utiliser les glucides, tandis que celles des communautés entériques portent leur préférence sur l'hydrolyse des lipides et protéines et 1'assimilation des acides gras, acides dicarboxyl iques, hydro'xy-acides et certains acides aminés. D'autre part les microflores entériques possèdent une versatilité nutritionnelle plus importante qui, liée à leurs effectifs, offre une potentialité catabolique non négligeable. .../... -2- Les mesures d'activité hétérotrophe effectuées au moyen de traceurs glucose 14 C permettent de montrer que les activités métaboliques des communautés bactériennes des sédiments abyssaux superficiels varient selon les zones. Dans le Cone de l'Amazone on observe un net gradient de minéralisation du glucose sur une distance de quelques kilomètres. Dans la plaine abyssale de Demerara les activités hétérotrophessont ralenties par rapport à celles des sédiments superficiels du Cone de l'Amazone, bien que les effectifs bactériens et les profondeurs soient du même ordre de grandeur dans ces deux zones. Les abondantes microflores associées aux différentes fractions du contenu du tractus digestif de l'holothurie abyssale Deima ne manifestent que peu d'activités métaboliques vis à vis du glucose. Par contre les communautés . bactériennes de Pseudostichopus et P syahropotes manifestent des activités hétérotrophes très fortes, croissant de 1'oesophage vers le cloaque. Les résultats actuels sont trop fragmentaires pour permettre d'observer une éventuelle spécialisation physiologique des communautés bactériennes entëriques permettant de faciliter les processus de digestion chez l'hôte. Conclusion: dans le milieu abyssal, caractérisé par des communautés bactériennes faiblement développées, les microflores associées aux contenus de tractus digestif des Echinodermes présentent des concentrations cellulaires considérablement plus importantes. Par leurs équipements enzymâtiques ces communautés entëriques complémentaires de ceux des microflores libres des sédiments superficiels, peuvent participer à la transformation des matières organiques du domaine abyssal. ETUDE DE LA DYNAMIQUE DES COMMUNAUTES MICROBIENNES ET DE LEURS ACTIVITES HETEROTROPHES DANS LES PARCS CONCHYLICOLES RESUME A partir d'expérimentations effectuées sur l'eau du bassin d'Arcachon, des essais de quantification et qualification du potentiel de réactivité des communautés bactériennes ont été effectuées. Les communautés bactériennes ont été définies par comptage directs, comptages après mise en culture, structure et diversité de ces communautés, dosage des AMP, ADP, ATP. Deux situations écologiques opposées ont étées considérées . Une situation automnale (température élevée) et une situation hivernale (température faible). Des bacs d'eau de mer de 6 et 60 litres ont été enrichis , soit avec du chlorure d'ammonium, soit avec un mélange d'acides aminés, à environ 500 uAtg N.l" . Les expériences ont duré 24 heures, avec des prélèvements toutes les 3 heures. La communauté automnale réagit rapidement (6 heures), la ' réponse à l'enrichissement en azote organique étant la plus importante. La structure de la communauté présente des stades immatures instables. Au contraire, au cours de l'expérimentation hivernale, aucune réponse n'a pu être perçue. Ceci peut signifier que la phase de latence est supérieure à 24 heures. La structure . de cette communauté présente des stades matures. En Conclusion, les communautés bactériennes réagissent de la même façon que les communautés macroscopiques. Les communautés stables sont diversifiées et correspondent à des stades matures de l'écosystème, les stades immatures sont instables et moins diversifiés. Ce type d'expérience a permi de mettre en évidence la rapidité de réponse des populations bactériennes, et peut être utilisé pour définir la stabilité d'une communauté bactérienne dans un biotope précis.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/1981/rapport-1926.pdf
http://archimer.ifremer.fr/doc/00000/1926/

Éditeur(s) : HAL CCSD American Association for Cancer Research
Résumé : International audience
The aim of this study was to test in vitro the efficacy of TAC, an original G-quadruplex ligand, as a potential radiosensitizing agent for glioblastoma multiforme (GBM). Two human radioresistant telomerase-positive GBM cell lines (SF763 and SF767) were analyzed, with and without TAC treatment, for telomere length, cell proliferation, apoptosis, cell-cycle distribution, gene expression, cytogenetic aberrations, clonogenic survival assay, 53BP1 immunofluorescence staining, and γH2AX phosphorylation. We found that low concentrations of TAC (0.5 and 1 μmol/L) inhibited the proliferation of GBM cells in a concentration-dependent manner after only 1 week of treatment, with minimal effects on cell cycle and apoptosis. TAC treatment had no visible effect on average telomere length but modified expression levels of telomere-related genes (hTERT, TRF1, and TRF2) and induced concentration-dependent DNA damage response and dicentric chromosomes. Survival curves analysis showed that exposure to nontoxic, subapoptotic concentrations of TAC enhanced radiation-induced killing of GBM cells. Analysis of DNA repair after irradiation revealed delayed repair kinetics in GBM cells treated with TAC. Furthermore, the combined treatment (TAC and radiation) significantly increased the frequency of chromosomal aberrations as compared with radiation alone. These findings provide the first evidence that exposure to a G4 ligand radiosensitizes human glioblastoma cells and suggest the prospect of future therapeutic applications.
ISSN: 1535-7163

hal-00851184
https://hal.archives-ouvertes.fr/hal-00851184
DOI : 10.1158/1535-7163.MCT-10-0664
PUBMED : 21987532

Extrait de : 2e journée d'études scientifiques des jeunes doctorants, le 21 avril 2016. Université des Antilles
Description : Alejandra PAGGI POLLI (doctorante en science politique, Université des Antilles) ouvre la réflexion sur la diplomatie territoriale et s'interroge sur l'intégration des territoires insulaires dans les grands espaces de décision. Il apparaît nécessaire de repenser le contexte international pour expliquer la configuration du nouveau système des relations internationales, dont ses traditionnels protagonistes, les Etats, ont été dépassés dans un monde globalisé. Les États modernes du modèle westphalien ont subi une série des transformations, surtout après la deuxième Guerre Mondiale, avec la prolifération d'organismes multilatérales comme l'ONU et l'UE qui ont provoqué progressivement un éloignement de leur position central et une cession de leur souveraineté en faveur de structures supranationales.

Droits : CC-BY-NC-ND - Attribution - Pas d'utilisation commerciale - Pas de modification
Permalien : http://www.manioc.org/fichiers/V16070
V16070

Éditeur(s) : HAL CCSD
Résumé : Background and objectives.[br/] Dairy propionibacteria are isolated from various ecological niches, including soil, grass, silage, rumen, raw milk and dairy plants, showing great adaptability and robustness. Used as ripening starters in Swiss type cheeses, these food grade bacteria are described as nutraceutical producers and they release into the external medium short chain fatty acids (SCFA), folic acid, cobalamin and the bifidogenic 1,4- dihydroxy-2-naphthoic acid (DHNA). These compounds are known to play a pivotal role in the modulation of intestinal physiology through diet. Considering the pro-apoptotic potential of some of these metabolites, we investigated the potential of dairy products fermented by the main species, Propionibacterium freudenreichii, to prevent colon carcinogenesis. Methods.[br/] We developed a new fermented milk. Taking advantage of milk fractions, high and stable populations of P. freudenreichii were obtained in this product. It was tested in vitro on gastric HGT-1 and colic HT-29 cells with respect to the typical hallmarks of apoptosis. In vivo, the fermented milk was evaluated in piglets with respect to general health parameters and in human microbiota-associated rats, treated or not with dimethylhydrazine (DMH), with respect to intestinal proliferation and apoptosis. Results. Fermented milk induced cell death in both gastric and colon cancer cells. Cellular and molecular characterization of cells dying in response to SCFA treatment revealed cell cycle arrest, drop in intracellular ATP, depolarization of mitochondria, translocation of key apoptosis proteins, processing of caspases and fragmentation of the nucleus. Interestingly, the new fermented milk was shown to potentiate the cytotoxic effect of molecules used in cancer chemotherapy. In accordance, it modulated expression of key apoptotic proteins. In vivo, the fermented milk was evaluated in piglets. Daily oral gavage led to high colic P. freudenreichii populations, to modulation of the intestinal microbiota and modulation of intestinal cytokines. Promotion of piglets’ growth also revealed general probiotic effects of fermented milk. Considering that propionibacteria were metabolically active in the intestine, we further investigated their impact on colon carcinogenesis (induced by dimethylhydrazine, DMH) in human microbiota-associated rats. Consumption of selected strains of propionibacteria resulted in enhanced apoptotic depletion of DMH damaged epithelial cells, yet had no effect on neither proliferation nor apoptosis in healthy rats. Conclusions and perspectives.[br/] These results open new perspectives in the field of colon cancer cells prevention and/or treatment. The synergy with pro-apoptotic chemotherapy molecules suggests that such a fermented product may be proposed as a food supplement to enhance the effects of anticancer treatments.
colloque international « Nutrition and Cancer : Hot topics from biology to public health issues »
Paris, France

hal-01190452
https://hal.archives-ouvertes.fr/hal-01190452
PRODINRA : 201853

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
Mechanical forces influence the growth and shape of virtually all tissues and organs. Recent studies show that increased cell contractibility, growth and differentiation might be normalized by modulating cell tensions. Particularly, the role of these tensions applied by the extracellular matrix during liver fibrosis could influence the hepatocarcinogenesis process. The objective of this study is to determine if 3D stiffness could influence growth and phenotype of normal and transformed hepatocytes and to integrate extracellular matrix (ECM) stiffness to tensional homeostasis. We have developed an appropriate 3D culture model: hepatic cells within three-dimensional collagen matrices with varying rigidity. Our results demonstrate that the rigidity influenced the cell phenotype and induced spheroid clusters development whereas in soft matrices, Huh7 transformed cells were less proliferative, well-spread and flattened. We confirmed that ERK1 played a predominant role over ERK2 in cisplatin-induced death, whereas ERK2 mainly controlled proliferation. As compared to 2D culture, 3D cultures are associated with epithelial markers expression. Interestingly, proliferation of normal hepatocytes was also induced in rigid gels. Furthermore, biotransformation activities are increased in 3D gels, where CYP1A2 enzyme can be highly induced/activated in primary culture of human hepatocytes embedded in the matrix. In conclusion, we demonstrated that increasing 3D rigidity could promote proliferation and spheroid developments of liver cells demonstrating that 3D collagen gels are an attractive tool for studying rigidity-dependent homeostasis of the liver cells embedded in the matrix and should be privileged for both chronic toxicological and pharmacological drug screening.
ISSN: 0730-2312

hal-01196600
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01196600
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01196600/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01196600/file/Increasing%203D%20Matrix%20Rigidity_accepted.pdf
DOI : 10.1002/jcb.25356

Éditeur(s) : Université des Antilles Études caribéennes
Résumé : La prolifération récente des résidences secondaires, observée récemment sur la côte Pacifique Sud du Nicaragua, s’inscrit dans une perspective de développement ayant trait à l’hébergement touristique. Or, le caractère immobilier prédominant de ce type d’hébergement, ainsi que les modes de consommation de ces résidents temporaires nous mettent en garde quant aux risques socioéconomiques et environnementaux encourus par les communautés autochtones. Parmi ces risques, notons les problèmes découlant de l’amplitude des variations saisonnières, de privatisation et de destruction des espaces naturels, ainsi que les perspectives de pérennisation des retombées économiques à moyen et long terme. Les auteurs de cet article remettent donc en question les prouesses attribuées à ce secteur d’activité et invitent à considérer des alternatives de développement durable du tourisme plus respectueuses des territoires d’accueil.
In recent years, there has been a proliferation of second home developments on attractive coastal regions of Central and South America. Based on the assumed benefits of second home tourism to regional economies, public authorities - accompanied by private investors – have capitalised on this sector as part of their tourism development strategies. This paper first examines the nature of recent second home tourism developments and then presents the socio-economic and environmental impacts of this phenomenon as experienced by local communities on the South Pacific Coast of Nicaragua. The main impacts include conflicts with local and indigenous communities over land use and ownership, seasonal and low-income employment generation, an increased burden on municipal budget to provide public infrastructure, and environmental degradation. It is argued that the second home tourism sector, strongly driven by private real estate investors, fails to generate tourism activities which are expected to sustain community development. Furthermore, this study indicates that municipalities concentrating on the second home tourism segment may deprive themselves of access to resources for more sustainable forms of tourism activities.
Amérique centrale
Nicaragua

Droits : info:eu-repo/semantics/openAccess
urn:doi:10.4000/etudescaribeennes.3869
http://journals.openedition.org/etudescaribeennes/3869

Éditeur(s) : HAL CCSD Paris : Muséum national d'histoire naturelle
Résumé : International audience
Coral reefs are one of the most diverse ecosystems on Earth and sustain millions of people worldwide.However, coral reefs are temporally dynamic and fragile systems subject to ever increasing disturbancesof anthoropogenic and natural origins. Understanding the patterns and trajectories of changes through time isurgently needed to estimate the resilience of coral reefs. To ascertain these patterns, we depend on the availabilityof long-term datasets, which are seldom available. In this study, we present the data set which allowed us to analysethe long-term trends of coral and fish assemblages for the island of Moorea (French Polynesia) from 1983to 2014, a dataset spanning 32 years, comprising two Acanthaster planci outbreaks (1987 and 2006) and severalhurricanes (1982, 1983, 1991 and 2010), on three permanent monitoring sites (quadrats of 50 × 2 m2) located onboth fringing and barrier reefs as well as on the outer slope. Coral cover oscillated significantly over time, particularlyon the outer slope where it suffered greatly from the effects of A. planci outbreaks and hurricanes. Totalfish and commercial fish abundances varied significantly across the three habitats. Prior to 2000, both total fishand herbivorous fish abundances increased on the barrier reef while after this time, numbers began to drop significantlyalong the outer slope. Species richness for total, commercialized and herbivorous fish increased over thesampling period for all three habitats. There were varied responses amongst species to disturbances on the outerslope over the sampling period: while some tended to disappear for a while before reappearing, others showed apeak in abundance in the middle of the sampling period, and still others increased or decreased continuously orwere stable over time. Our results support the view that coral reefs are highly dynamic systems and that differentfunctional groups show different temporal trajectories. While disturbance is probably the major driver for coralcover dynamics, its impact is less obvious for fish, possibly due to their mobility and population dynamics whichmay mask the effect of disturbance.
Les récifs coralliens sont des systèmes dynamiques et diversifiés régulièrement soumis à des perturbationsnaturelles et anthropiques, sources d’hétérogénéité spatiale et temporelle dans la structure et la dynamique deces communautés d’organismes que sont, entre autres, les poissons et les coraux. Récemment, il a été démontréque les activités humaines pouvaient modifier les régimes naturels de perturbations des récifs coralliens en transformantles événements ponctuels en perturbations persistantes ou en stress chronique, soit en introduisant denouveaux types de perturbations, soit en ajoutant ou altérant certaines perturbations naturelles nécessaires au bonmaintien de la dynamique inhérente aux récifs. Dans cette étude, nous présentons la base de données qui nouspermettra d’analyser les tendances à long-terme des assemblages de poissons et de coraux de l’île de Moorea(Polynésie française) entre 1983 et 2014 sur une période de 32 ans, comprenant deux proliférations d’Acanthasterplanci (1987 et 2006) et plusieurs cyclones (1982, 1983, 1991 et 2010), sur trois sites de suivi permanents(quadrats de 50 × 2 m2) sur les récifs barrière et frangeant ainsi que sur la pente externe. La couverture corallienneprésente de fortes oscillations, en particulier sur la pente externe où elle a répondu fortement à la proliférationd’A. planci et des cyclones. Les abondances des poissons totaux et des poissons commercialisés montrent destendances contrastées sur les trois habitats, avec, plus particulièrement, une augmentation sur le récif barrière etune diminution sur la pente externe après 2000. Ces tendances se retrouvent chez les poissons herbivores dansces deux habitats. La richesse spécifique des poissons totaux, commerciaux et herbivores, augmente de façoncontinue au cours de la période d’étude dans les trois habitats. Des groupes particuliers d’espèces répondentdifféremment aux perturbations de la pente externe au cours du suivi : alors que certaines espèces ont tendanceà disparaître pendant quelques années avant de réapparaître, d’autres présentent un pic en terme d’abondanceen milieu de suivi ; d’autres espèces ont des abondances qui augmentent ou diminuent de façon continue, etd’autres encore montrent des abondances stables. Ceci indique que les récifs coralliens sont des systèmes trèsdynamiques dans lesquels les perturbations naturelles peuvent affecter les espèces de façon différente. Alors quele régime de perturbations naturelles semble être important pour la dynamique de la couverture corallienne danscertains habitats, cela se retrouve beaucoup moins pour les poissons. Cette atténuation des fluctuations spatialeset temporelles peut être due aux aptitudes de dispersion de nombreuses espèces de poissons, capables de chercherun autre habitat favorable en cas de perturbation de l’habitat corallien d’origine.
ISSN: 0399-0974

hal-01282378
https://hal-univ-perp.archives-ouvertes.fr/hal-01282378

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
Cytochrome P450 aromatase (P450arom; aromatase) is a microsomal enzyme involved in the production of endogeneous sex steroids by converting testosterone into oestradiol. Aromatase is the product of the cyp19a1 gene and plays a crucial role in the sexual differentiation of the brain and in the regulation of reproductive functions. In the brain of mammals and birds, expression of cyp19a1 has been demonstrated in neuronal populations of the telencephalon and diencephalon. By contrast, a wealth of evidence established that, in teleost fishes, aromatase expression in the brain is restricted to radial glial cells. The present study investigated the precise neuroanatomical distribution of cyp19a1 mRNA during brain development in Xenopus laevis (late embryonic to juvenile stages). For this purpose, we used in situ hybridisation alone or combined with the detection of a proliferative (proliferating cell nuclear antigen), glial (brain lipid binding protein, Vimentin) or neuronal (acetylated tubulin; HuC/D; NeuroβTubulin) markers. We provide evidence that cyp19a1 expression in the brain is initiated from the very early larval stage and remains strongly detected until the juvenile and adult stages. At all stages analysed, we found the highest expression of cyp19a1 in the preoptic area and the hypothalamus compared to the rest of the brain. In these two brain regions, cyp19a1-positive cells were never detected in the ventricular layers. Indeed, no co-labelling could be observed with radial glial (brain lipid binding protein, Vimentin) or dividing progenitors (proliferating cell nuclear antigen) markers. By contrast, cyp19a1-positive cells perfectly matched with the distribution of post-mitotic neurones as shown by the use of specific markers (HuC/D, acetylated tubulin and NeuroβTubulin). These data suggest that, similar to that found in other tetrapods, aromatase in the brain of amphibians is found in post-mitotic neurones and not in radial glia as reported in teleosts.
ISSN: 0953-8194

hal-00994513
https://hal-univ-rennes1.archives-ouvertes.fr/hal-00994513
DOI : 10.1111/jne.12142
PUBMED : 24612124

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
CXCL-test is a method that uses the estrogen-dependent secretion of the natural endogenous chemokine CXCL12 to evaluate the estrogenic activity of molecules. CXCL12 chemokine is involved in the estrogen dependent proliferation of breast cancer cells. Its measure is an indicator of cell proliferation and is used as an alternative test to classical proliferation test. Here we aimed to optimize this test, first to increase the number of tested molecules in a single assay and then to decrease the number of intermediate steps. The optimized CXCL-test was finally used for the evaluation of the estrogenic potency of emerging chemical pollutants: the UV filter benzophenones (BPs). The effect of BPs on CXCL12 secretion was also validated by real time quantitative RT-PCR. The optimized CXCL-test allowed a fast and direct assessment of estrogenic potency of molecules. The estrogenic activities of benzophenones were characterized and divided in two groups. The first one contains weak estrogenic compounds (BP, BP1, BP2, BP3, 234BP and 2344′BP). The second one contains medium estrogenic compounds (4BP, 44′BP, BP8, THB).
ISSN: 0045-6535

hal-01477205
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477205
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477205/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477205/file/Rapid%20assessment%20of%20estrogenic%20compounds_accepted.pdf
DOI : 10.1016/j.chemosphere.2017.01.037

Éditeur(s) : HAL CCSD MDPI
Résumé : International audience
In mammals, the effects of estrogen are mainly mediated by two different estrogen receptors, ERα and ERβ. These proteins are members of the nuclear receptor family, characterized by distinct structural and functional domains, and participate in the regulation of different biological processes, including cell growth, survival and differentiation. The two estrogen receptor (ER) subtypes are generated from two distinct genes and have partially distinct expression patterns. Their activities are modulated differently by a range of natural and synthetic ligands. Some of these ligands show agonistic or antagonistic effects depending on ER subtype and are described as selective ER modulators (SERMs). Accordingly, a few phytochemicals, called phytoestrogens, which are synthesized from plants and vegetables, show low estrogenic activity or anti-estrogenic activity with potentially anti-proliferative effects that offer nutraceutical or pharmacological advantages. These compounds may be used as hormonal substitutes or as complements in breast cancer treatments. In this review, we discuss and summarize the in vitro and in vivo effects of certain phytoestrogens and their potential roles in the interaction with estrogen receptors.
ISSN: 1422-0067

hal-01560283
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01560283
PUBMED : 28657580
DOI : 10.3390/ijms18071381

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Xeno-estrogens, a class of endocrine disrupting chemicals (EDCs), can disturb estrogen receptor-dependent pathways involved in differentiation, proliferation or protection. Multiple methods have been developed to characterize the disturbances induced by EDCs in different cells or organs. In this study we have developed a new tool for the assessment of estrogenic compounds on differentiation. For this purpose we used the global model of NGF-induced neurite outgrowth of a pseudoneuronal PC12 cell line stably transfected with estrogen receptor alpha (PC12 ER). This new test evidences a new selectivity in which estradiol, genistein and 4-hydroxytamoxifen increased the NGF-induced neurite outgrowth of PC12 ER cells in a dose-dependent manner. In contrast, the strong estrogen agonist 17α-ethynylestradiol, the strong antagonist raloxifene and the agonist bisphenol A were unable to modify the neuritogenesis of PC12 ER cells. Therefore, the analysis of neuritogenesis in PC12 ER cells constitutes a complementary tool for the characterization of xeno-estrogen activity and also serves as a basis for further studies focusing on the mechanisms of EDCs in a neuronal context. Moreover, this test constitutes an alternative to animal testing.
ISSN: 0045-6535

hal-01063907
https://hal.archives-ouvertes.fr/hal-01063907
https://hal.archives-ouvertes.fr/hal-01063907/document
https://hal.archives-ouvertes.fr/hal-01063907/file/Differentiationsur PC12_cells_accepted.pdf
DOI : 10.1016/j.chemosphere.2014.03.101

Éditeur(s) : HAL CCSD
Résumé : International audience
Primary cultures of hepatocytes are powerful models in studying the sequence of events that are necessary for cell progression from a G0-like state to S phase. The models mimic the physiological process of hepatic regeneration after liver injury or partial hepatectomy. Many reports suggest that the mitogen-activated protein kinase (MAPK) ERK1/2 can support hepatocyte proliferation in vitro and in vivo and the MEK/ERK cascade acts as an essential element in hepatocyte responses induced by the EGF. Moreover, its disregulation has been associated with the promotion of tumor cell growth of a variety of tumors, including hepatocellular carcinoma. Whereas the strict specificity of action of ERK1 and ERK2 is still debated, the MAPKs may have specific biological functions under certain contexts and according to the differentiation status of the cells, notably hepatocytes. In this paper, we will focus on MEK1/2-ERK1/2 activations and roles in normal rodent hepatocytes in vitro and in vivo after partial hepatectomy and in human hepatocarcinoma cells. The possible specificity of ERK1 and ERK2 in normal and transformed hepatocyte will be discussed in regard to other differentiated and undifferentiated cellular models.
International Journal of Hepatology

Droits : info:eu-repo/semantics/OpenAccess
hal-00875664
https://hal.archives-ouvertes.fr/hal-00875664
https://hal.archives-ouvertes.fr/hal-00875664/document
https://hal.archives-ouvertes.fr/hal-00875664/file/TheMAPKMEK12-accepted.pdf
DOI : 10.1155/2012/328372

Éditeur(s) : HAL CCSD BioMed Central
Résumé : International audience
Background: Myosin II (or Myosin Heavy Chain II, MHCII) is a family of molecular motors involved in the contractile activity of animal muscle cells but also in various other cellular processes in non-muscle cells. Previous phylogenetic analyses of bilaterian MHCII genes identified two main clades associated respectively with smooth/non-muscle cells (MHCIIa) and striated muscle cells (MHCIIb). Muscle cells are generally thought to have originated only once in ancient animal history, and decisive insights about their early evolution are expected to come from expression studies of Myosin II genes in the two non-bilaterian phyla that possess muscles, the Cnidaria and Ctenophora. Results: We have uncovered three MHCII paralogues in the ctenophore species Pleurobrachia pileus. Phylogenetic analyses indicate that the MHCIIa /MHCIIb duplication is more ancient than the divergence between extant metazoan lineages. The ctenophore MHCIIa gene (PpiMHCIIa) has an expression pattern akin to that of ``stem cell markers'' (Piwi, Vasa...) and is expressed in proliferating cells. We identified two MHCIIb genes that originated from a ctenophore-specific duplication. PpiMHCIIb1 represents the exclusively muscular form of myosin II in ctenophore, while PpiMHCIIb2 is expressed in non-muscle cells of various types. In parallel, our phalloidin staining and TEM observations highlight the structural complexity of ctenophore musculature and emphasize the experimental interest of the ctenophore tentacle root, in which myogenesis is spatially ordered and strikingly similar to striated muscle formation in vertebrates. Conclusion: MHCIIa expression in putative stem cells/proliferating cells probably represents an ancestral trait, while specific involvement of some MHCIIa genes in smooth muscle fibres is a uniquely derived feature of the vertebrates. That one ctenophore MHCIIb paralogue (PpiMHCIIb2) has retained MHCIIa-like expression features furthermore suggests that muscular expression of the other paralogue, PpiMHCIIb1, was the result of neofunctionalisation within the ctenophore lineage, making independent origin of ctenophore muscle cells a likely option.
ISSN: 1471-2148

hal-01544486
https://hal.archives-ouvertes.fr/hal-01544486
DOI : 10.1186/1471-2148-12-107

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
The results from recent studies show that some benzophenones (BPs) and their hydroxylated metabolites can function as weak estrogens (E2) in the environment. However, little is known about the structure-activity relationship of these molecules. We have examined the effects of exposure to ten different BPs on the proliferation of estrogen receptor (ER)-positive breast cancer cells and on the transcriptional activity of E2-target genes. We analyzed two genes that are tightly linked with estrogen-mediated proliferation, the CXCL12 and amphiregulin genes and two classical estrogen-responsive genes, the pS2 and progesterone receptor. Significant differences in the BPs efficiency to induce cell proliferation and endogenous E2-target gene expressions were observed. Using ERE-, Sp1-, AP1- and C3-reporter genes that contain different ER-binding sites in their promoter, we also showed significant differences in the BPs efficiency in activation of the ER transactivation. Together, our analyzes showed that the most active molecule is 4-hydroxy-BP. Docking analysis of the interaction of BPs in the ligand-binding pocket of ERα suggests that the minimum structural requirement for the estrogenic activity of BPs is a hydroxyl (OH) group in the phenyl A-ring that allows interaction with Glu-353, Arg-394 or Phe-404, which enhances the stability between BPs and ERα. Our modeling also indicates a loss of interaction between the OH groups of the phenyl B-ring and His-524. In addition, the presence of some OH groups in the phenyl B-ring can create repulsion forces, which may constrain helix 12 in an unfavorable position, explaining the differential estrogenic effects of BPs. These results, together with our analysis of BPs for their potency in activation of cell proliferation and ER-mediated transcription, report an improved understanding of the mechanism and structure-activity relationship of BPs.
ISSN: 1932-6203

inserm-00838323
http://www.hal.inserm.fr/inserm-00838323
http://www.hal.inserm.fr/inserm-00838323/document
http://www.hal.inserm.fr/inserm-00838323/file/journal.pone.0060567.pdf
DOI : 10.1371/journal.pone.0060567

Résumé : Suite au Sommet de Rio en 1992, des réflexions ont abouti à la mise en place de nombreux programmes (nationaux et internationaux) concernant l'étude des milieux modifiés par les actions anthropiques. Parmi eux, le programme national "Dynamique de la Biodiversité et Environnement", auquel participe le laboratoire DEL/Ecologie du Centre IFREMER de Brest, se penche sur le problème des espèces proliférantes en milieu terrestre et marin. Dans ce deuxième cas, les exemples sont peu nombreux mais on citera tout de même les proliférations provoquées par un déséquilibre du milieu (marées vertes (ulves)) et celles dues à l'introduction volontaire ou accidentelle d'espèces (sargasses, caulerpes, crépidules). Crepidula fornicata fait ainsi l'objet d'études particulières de la part du laboratoire d'écologie benthique. Cette espèce présente en effet des potentialités impressionnantes de colonisation de toutes sortes de milieux (baies, estuaires, secteurs conchylicoles) provoquant des déséquilibres dans les écosystèmes envahis. Le programme a donc pour objectifs l'identification du processus de colonisation (incluant l'évaluation de la responsabilité de l'Homme), l'étude des effets sur les populations voisines, et la caractérisation des interactions entre la crépidule et les mollusques filtreurs exploités.

Droits : Ifremer
http://archimer.ifremer.fr/doc/00014/12553/9419.pdf
http://archimer.ifremer.fr/doc/00014/12553/

Éditeur(s) : HAL CCSD BioMed Central
Résumé : International audience
BACKGROUND: Hepatocellular carcinoma (HCC) is a particularly severe disease characterized by a high rate of recurrence and death even after surgical resection. Molecular characterization of HCC helps refine prognosis and may facilitate the development of improved therapy. Phosphatidylinositol 4-kinases have recently been identified as cellular factors associated with cancer. Also, phosphatidylinositol 4-kinase type IIIα (PI4KA) is necessary for the propagation of the hepatitis C virus, a major etiological factor for HCC. METHODS: Reverse transcription, quantitative real-time PCR was used to assay PI4KA mRNA. The expression levels were investigated both in a collection of molecularly and clinically characterized hepatic tissues from 344 patients with diverse liver diseases and in human hepatocyte cell lines whose proliferative and differentiation status was controlled by specific culture conditions. Analytical microarray data for 60 HCC and six normal liver tissue samples were exploited to study correlations between PI4KA mRNA levels and cell proliferation markers in vivo. Postoperative disease-specific survival and time to recurrence in a set of 214 patients with HCC were studied by univariate and multivariate analyses. RESULTS: PI4KA mRNA was more abundant in HCC than normal healthy tissues. This upregulation correlated significantly with both poor differentiation and the active proliferation rate in HCC. These associations were confirmed with in vitro models. Moreover, patients with HCC who had been treated by surgical resection and had higher PI4KA mRNA concentrations in their tumor tissue exhibited a higher risk of tumor recurrence (median time: 20 months versus 49 months, P = 0.0012) and shorter disease-specific survival (first quartile time: 16 months versus 48 months, P = 0.0004). Finally, the abundance of PI4KA mRNA proved to be an independent prognostic marker of survival for cases of HCC (hazard ratio = 2.36, P = 0.0064). CONCLUSIONS: PI4KA mRNA could be used as a new molecular marker to improve established prognostic models for HCC. These findings also indicate possible new lines of research for the development of innovative therapeutic approaches targeting PI4KA.
ISSN: 1471-2407

inserm-00934294
http://www.hal.inserm.fr/inserm-00934294
http://www.hal.inserm.fr/inserm-00934294/document
http://www.hal.inserm.fr/inserm-00934294/file/1471-2407-14-7.pdf
DOI : 10.1186/1471-2407-14-7

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
The cyp19a1 gene that encodes aromatase, the only enzyme permitting conversion of C19 aromatizable androgens into estrogens, is present as a single copy in the genome of most vertebrate species, except in teleosts in which it has been duplicated. This study aimed at investigating the brain expression of a cyp19a1 gene expressed in both gonad and brain of Japanese eel, a basal teleost. By means of immunohistochemistry and in situ hybridization, we show that cyp19a1 is expressed only in radial glial cells of the brain and in pituitary cells. Treatments with salmon pituitary homogenates (female) or human chorionic gonadotrophin (male), known to turn on steroid production in immature eels, strongly stimulated cyp19a1 messenger and protein expression in radial glial cells and pituitary cells. Using double staining studies, we also showed that aromatase-expressing radial glial cells exhibit proliferative activity in both the brain and the pituitary. Altogether, these data indicate that brain and pituitary expression of Japanese eel cyp19a1 exhibits characteristics similar to those reported for the brain specific cyp19a1b gene in teleosts having duplicated cyp19a1 genes. This supports the hypothesis that, despite the fact that eels also underwent the teleost specific genome duplication, they have a single cyp19a1 expressed in both brain and gonad. Such data also suggest that the intriguing features of brain aromatase expression in teleost fishes were not gained after the whole genome duplication and may reflect properties of the cyp19a1 gene of ancestral Actinopterygians.
ISSN: 1932-6203

hal-00877375
https://hal.archives-ouvertes.fr/hal-00877375
https://hal.archives-ouvertes.fr/hal-00877375/document
https://hal.archives-ouvertes.fr/hal-00877375/file/Expressionsur Aromatase-accepted.pdf
DOI : 10.1371/journal.pone.0044750

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
Recent reports suggest that extracellular signal-regulated kinase (ERK1) and ERK2 mitogen-activated protein kinases (MAPK) may direct specific biological functions under certain contexts. In this study, we investigated the role of early and sustained epidermal growth factor (EGF) stimulation on long-term hepatocyte differentiation and the possible role of ERK1 and ERK2 in this process. We demonstrate a long-term survival and an elevated level of differentiation up to 3 weeks. The differentiation state of hepatocytes is supported by sustained expression of aldolase B, albumin, and the detoxifying enzymes CYP1A2, 2B2, and 3A23. Similarly to freshly isolated cells, cultured hepatocytes also retain the ability to respond to 3-methylcholanthrene (3MC) and phenobarbital (PB), two known CYP inducers. In addition, we show evidence that continuous MAPK/ERK kinase (MEK) inhibition enhances the level of differentiation. Using RNA interference approaches against ERK1 and ERK2, we demonstrate that this effect requires both ERK1 and ERK2 activity, whereas the specific ERK1 knockdown promotes cell survival and the specific ERK2 knockdown regulates cell proliferation. In conclusion, we demonstrate that early and sustained EGF stimulation greatly extends long-term hepatocyte survival and differentiation, and that inhibition of the ERK1/2 MAPK pathway potentiates these pro-survival/pro-differentiation phenotypes. We clearly attest that specific ERK1 and ERK2 MAPKs determine hepatocyte survival and proliferation, respectively, whereas dual inhibition is required to stabilize a highly differentiated state.
ISSN: 0021-9541

hal-00681056
https://hal.archives-ouvertes.fr/hal-00681056
DOI : 10.1002/jcp.22742
PUBMED : 21437905