Éditeur(s) : HAL CCSD Society for the Study of Reproduction
Résumé : International audience
Matrix-assisted laser desorption/ionization (MALDI) tissue imaging mass spectrometry is particularly promising among the numerous applications of mass spectrometry. It is used for probing and analyzing the spatial arrangement of a wide range of molecules, including proteins, peptides, lipids, drugs, and metabolites, directly in thin slices of tissue. In the field of proteomics, the technology avoids tedious and time-consuming extraction and fractionation steps classically required for sample analysis. MALDI imaging mass spectrometry is increasingly recognized as a powerful method for clinical proteomics, particularly in cancer research. The technology has particular potential for the discovery of new tissue biomarker candidates, classification of tumors, early diagnosis or prognosis, elucidating pathogenesis pathways, and therapy monitoring. Over recent years, MALDI imaging mass spectrometry has been used for molecular profiling and imaging directly in male and female reproductive tissues. This review will consider some of the recent publications in the field, addressing a range of issues covering embryo development, gene expression product profiling during gametogenesis, and seeking and identifying biomarkers of reproductive cancers. The wealth of advances in mass spectrometry imaging will inevitably attract biologists and clinicians as the advantages and power of this technology become more widely known. This review will also discuss bottlenecks and the many technical issues that remain to be resolved before laboratories in the field can adopt the technology. We foresee that MALDI imaging mass spectrometry will have a major impact in reproductive research by opening new avenues to the understanding of various molecular mechanisms and the diagnosis of reproductive pathologies.
ISSN: 0006-3363

hal-00877851
https://hal.archives-ouvertes.fr/hal-00877851
DOI : 10.1095/biolreprod.111.094896
PUBMED : 22156474

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Accumulating evidence suggests that developmental exposure to environmental chemicals may modify the course of brain development, ultimately leading to neuropsychiatric / neurodegenerative disorders later in life. In the present study, we assessed the impact of one of the most frequently used pesticides in both residential and agricultural applications - the synthetic pyrethroid cypermethrin (CYP) - on developmental neurotoxicity (DNT). Female mice were perinatally exposed to low doses of CYP (5 and 20 mg/kg body weight) from gestation to postnatal day 15. Behavioral analyses were performed during the offspring's early life and during adulthood. Postnatal analyses revealed that perinatal exposure to CYP disturbed motor development without modifying sensory and communicative skills. We found that later in life, CYP-exposed offspring expressed maladaptive behaviors in response to highly challenging tasks and abnormal sociability. Transcriptomic analyses performed in the offspring's brain at the end of the exposure, highlighted mitochondrial dysfunction as a relevant pathomechanism underlying CYP-induced DNT. Interestingly, several genes involved in proteostasis maintenance were also shown to be dysregulated suggesting that alterations in biogenesis, folding, trafficking and degradation of proteins may significantly contribute to CYP-related DNT. From a regulatory perspective, this study highlights that behavioral and transcriptomic analyses are complementary tools providing useful direction for better DNT characterization, and as such, should be used together more systematically.
ISSN: 1932-6203

hal-01616489
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01616489
DOI : 10.1371/journal.pone.0184475
PUBMED : 29020013

Éditeur(s) : Elsevier
Résumé : in this study we evaluated the effects of the probiotic Pediococcus acidilactici MA18/5M on shrimp, Litopenaeus stylirostris (also called Penaeus stylirostris), first on antioxidant defences and secondly on the oxidative stress status in the shrimps' haemolymph and digestive gland. We conducted two experiments with the same protocol in which shrimps were fed two diets for three weeks: a control diet and a probiotic diet containing 1 g of live P. acidilactici MA18/5M kg(-1). In the first experiment, the shrimps were found to be healthy over the trial period; no mortalities and non-detectable signs of infection were recorded. These resulted in high final survival rates (above 90% in both treatments). On the other hand, during the second trial, carried out at a period of increased risk for an outbreak of the summer syndrome, higher mortalities were recorded, associated with high V. nigripulchritudo prevalence and loads in the shrimp haemolymph. In healthy shrimps (trial 1), no detectable response of the antioxidant defence system and of oxidative stress bio-indicators were observed. However, feeding the probiotic significantly increased the total antioxidant status (TAS) and glutathione peroxidase activity, while all other parameters remained significantly unchanged. In the shrimps exposed to V nigripulchritudo (trial 2), the antioxidant response was characterized by higher antioxidant enzyme activities (superoxide dismutase and catalase) and higher oxidative stress level in the digestive gland (higher oxidized/reduced glutathione ratio, higher malonclialdehyde and carbonyl protein contents) compared to levels found in trial 1. However, shrimps fed the probiotic diet exhibited (i) significantly higher final survival rates (67 +/- 3%) compared to the control (47 +/- 4%) and (ii) a lower prevalence of V nigripulchritudo throughout the trial. Moreover, the antioxidant response and the oxidative stress level recorded in the digestive gland with shrimps submitted to the probiotic diet were lower. In the light of these results, we confirm that P acidilactici enrichment in shrimps' diet seems an effective way of reducing the susceptibility of shrimps to bacterial pathogens. (c) 2009 Elsevier B.V. All rights reserved.
Aquaculture (0044-8486) (Elsevier), 2009-09 , Vol. 294 , N. 3-4 , P. 306-313

Droits : 2009 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/2009/publication-6780.pdf
DOI:10.1016/j.aquaculture.2009.06.016
http://archimer.ifremer.fr/doc/00000/6780/

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Genetics and molecular biology have been instrumental for a better understanding of heritable defects causing human infertility over the past decades. More recently, the field of reproductive biology has harnessed genome biological approaches to gain insight into molecular processes underlying normal and pathological gametogenesis and gamete function. We are currently witnessing yet another quantum leap in our ability to monitor the flow of information from the genome via the transcriptome to the proteome: tiling arrays that cover both strands of a given target genome and RNA-Seq, a method based on ultra-high throughput DNA sequencing, enable us to study noncoding and protein-coding transcripts with unprecedented precision and depth at a reasonable cost. These technologies have spawned a thriving discipline within the bioinformatics field that employs information technology for managing and interpreting biological high-throughput data. This review outlines database projects and online analysis tools useful for life scientists in general and discusses in detail selected projects that have specifically been developed for researchers and clinicians in the field of reproductive biology. This article is part of a Special Issue entitled: Molecular Genetics of Human Reproductive Failure.
ISSN: 0006-3002

hal-00877444
https://hal.archives-ouvertes.fr/hal-00877444
DOI : 10.1016/j.bbadis.2012.05.018
PUBMED : 22687534

Éditeur(s) : HAL CCSD Springer New York
Résumé : International audience
Estrogen receptor α (ERα) exerts regulatory actions through genomic mechanisms. In the classical pathway, ligand-activated ERα binds directly to DNA through estrogen response elements (ERE) located in the promoter of target genes. ERα can also exert indirect regulation of transcription via protein-protein interaction with other transcription factors such as AP-1. Several ERα synonymous polymorphisms have been identified and efforts to understand their implications have been made. Nevertheless effects of synonymous polymorphisms are still neglected. This chapter focuses on the experimental procedure employed in order to characterize the transcriptional activity of a synonymous polymorphism of the ERα (rs746432) called Alanine 87 (Ala87). Activity of both WT and Ala87 ERα isoforms on transcriptional pathways can be analyzed in transiently transfected cells using different reporter constructs. ERα efficiency on the classical genomic pathway can be analyzed by determining its transactivation activity on an ERE-driven thymidine kinase (TK) promoter controlling the expression of the luciferase reporter gene. Transcriptional activity through the indirect genomic pathway can be analyzed by employing an AP-1 DNA response element-driven promoter also controlling the expression of luciferase reporter gene
Estrogen Receptors

ISBN : 978-1-4939-3126-2
hal-01237073
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01237073
DOI : 10.1007/978-1-4939-3127-9_22

Éditeur(s) : HAL CCSD Nature Publishing Group
Résumé : International audience
CYLD is a tumour suppressor gene mutated in familial cylindromatosis, a genetic disorder leading to the development of skin appendage tumours. It encodes a deubiquitinating enzyme that removes Lys63- or linear-linked ubiquitin chains. CYLD was shown to regulate cell proliferation, cell survival and inflammatory responses, through various signalling pathways. Here we show that CYLD localizes at centrosomes and basal bodies via interaction with the centrosomal protein CAP350 and demonstrate that CYLD must be both at the centrosome and catalytically active to promote ciliogenesis independently of NF-κB. In transgenic mice engineered to mimic the smallest truncation found in cylindromatosis patients, CYLD interaction with CAP350 is lost disrupting CYLD centrosome localization, which results in cilia formation defects due to impairment of basal body migration and docking. These results point to an undiscovered regulation of ciliogenesis by Lys63 ubiquitination and provide new perspectives regarding CYLD function that should be considered in the context of cylindromatosis.
ISSN: 2041-1723

hal-01063915
https://hal.archives-ouvertes.fr/hal-01063915
DOI : 10.1038/ncomms5585
PUBMED : 25134987

Résumé : Tahiti’s pearl farming industry plays a major socio-economic role in French Polynesia. In an increasingly competitive market where the production of high quality pearls becomes essential, research can help secure and ensure sustainable production. In that context, Ifremer, in close collaboration with the “direction des resources marines” (French Polynesian government agency) has developed research projects on the “sustainable development of pearl farming”. This program is organized along 3 axes: (1) understanding the animal physiology and initiating a genetically selective breeding program of donor oysters; (2) understanding pearl oyster larvae dispersal and recruitment; (3) understanding the molecular mechanisms underlying biomineralization processes during shell and pearl formation. It is in this frame that we have developed a highthroughput Expressed Sequence Tags pyrosequencing program on the calcifying mantle, combined with proteomic analyses of the shell and pearl. We analyzed 276738 EST sequences, leading to the constitution of a P. margaritifera mantle transcripts catalogue of 82 sequences potentially involved in biomineralization. Our results provided direct evidence that our ESTs data set covers a large number of the matrix proteins of P. margaritifera. In addition, our proteomic analysis enabled us to retrieve, in silico, all the sequences from P. margaritifera involved in the biomineralization process already published on databases. Integration of these two methods allowed, for the first time, the global composition of calcifying tissue and calcified structures to be examined in tandem.
Recent Advances in Pearl Research—Proceedings of the International Symposium on Pearl Research 2011.Edited by Shugo Watabe, Kaoru Maeyama and Hiromichi Nagasawa. 2013. ISBN : 978-4-88704-158-5. pp.184-194

Droits : 2013 TERRAPUB. All rights reserved.
http://archimer.ifremer.fr/doc/00167/27840/26049.pdf
http://archimer.ifremer.fr/doc/00167/27840/

Éditeur(s) : HAL CCSD BioMed Central
Résumé : International audience
Background: Microbial genetic diversity is often investigated via the comparison of relatively similar 16S molecules through multiple alignments between reference sequences and novel environmental samples using phylogenetic trees, direct BLAST matches, or phylotypes counts. However, are we missing novel lineages in the microbial dark universe by relying on standard phylogenetic and BLAST methods? If so, how can we probe that universe using alternative approaches? We performed a novel type of multi-marker analysis of genetic diversity exploiting the topology of inclusive sequence similarity networks. Results: Our protocol identified 86 ancient gene families, well distributed and rarely transferred across the 3 domains of life, and retrieved their environmental homologs among 10 million predicted ORFs from human gut samples and other metagenomic projects. Numerous highly divergent environmental homologs were observed in gut samples, although the most divergent genes were over-represented in non-gut environments. In our networks, most divergent environmental genes grouped exclusively with uncultured relatives, in maximal cliques. Sequences within these groups were under strong purifying selection and presented a range of genetic variation comparable to that of a prokaryotic domain. Conclusions: Many genes families included environmental homologs that were highly divergent from cultured homologs: in 79 gene families (including 18 ribosomal proteins), Bacteria and Archaea were less divergent than some groups of environmental sequences were to any cultured or viral homologs. Moreover, some groups of environmental homologs branched very deeply in phylogenetic trees of life, when they were not too divergent to be aligned. These results underline how limited our understanding of the most diverse elements of the microbial world remains, and encourage a deeper exploration of natural communities and their genetic resources, hinting at the possibility that still unknown yet major divisions of life have yet to be discovered.
ISSN: 1745-6150

Droits : http://creativecommons.org/licenses/by/
hal-01257771
http://hal.upmc.fr/hal-01257771
http://hal.upmc.fr/hal-01257771/document
http://hal.upmc.fr/hal-01257771/file/s13062-015-0092-3.pdf
DOI : 10.1186/s13062-015-0092-3

Éditeur(s) : HAL CCSD Society for the Study of Reproduction
Résumé : International audience
Transcription factor GATA4 is required for the development and function of the mammalian gonads. We first reported that the GATA4 gene in both human and rodents is expressed as two major alternative transcripts that differ solely in their first untranslated exon (exon 1a vs. exon 1b). We had also showed by quantitative PCR that in mouse tissues, both Gata4 exon 1a- and 1b-containing transcripts are present in all sites that are normally positive for GATA4 protein. In adult tissues, exon 1a-containing transcripts generally predominate. A notable exception, however, is the testis where the Gata4 exon 1a and 1b transcripts exhibit a similar level of expression. We now confirm by in situ hybridization analysis that each transcript is also strongly expressed during gonad differentiation in both sexes in the rat. To gain further insights into how Gata4 gene expression is controlled, we characterized the mouse Gata4 promoter sequence located upstream of exon 1b. In vitro studies revealed that the Gata4 1b promoter is less active than the 1a promoter in several gonadal cell lines tested. Whereas we have previously shown that endogenous Gata4 transcription driven by the 1a promoter is dependent on a proximally located Ebox motif, we now show using complementary in vitro and in vivo approaches that Gata4 promoter 1b-directed expression is regulated by GATA4 itself. Thus, Gata4 transcription in the gonads and other tissues is ensured by distinct promoters that are regulated differentially and independently.
ISSN: 0006-3363

hal-01010626
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01010626
DOI : 10.1095/biolreprod.113.113290
PUBMED : 24352556

Éditeur(s) : Elsevier Science Bv
Résumé : In this study, three microdiets were tested on weaning of Florida pompano larvae: Otohime, Gemma and a reference diet LR803. The experimental system was stocked with 11-day-old larvae, which were co-fed micro-diets and live food from 11 dph to 17 dph then micro-diets only until 28 dph. Survival from 11 dph to 28 dph was similar for all treatments, with an average of 33%. At the end of the trial, the Gemma larvae were significantly longer and heavier than larvae fed the other diets. Significant differences were observed in fatty acid composition of the diets and larvae between treatments. The Gemma larvae incorporated the lowest amount of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and arachidonic acid (ARA). However, they had the highest DHA/EPA and ARA/EPA ratios, which is in agreement with the concept that the proportions of polyunsaturated fatty acids could be of greater importance than their absolute amount. Results from the enzyme analysis suggest that fishmeal is suitable as the main protein source for Florida pompano larvae compared to krill meal. This study gives new insights on Florida pompano early nutritional requirements and demonstrated the full functionality of the pancreas at 16 days post hatch, opening possibilities of an earlier weaning time.
Aquaculture (0044-8486) (Elsevier Science Bv), 2014-02 , Vol. 422 , P. 268-276

Droits : 2013 Elsevier B.V. All rights reserved
http://archimer.ifremer.fr/doc/00169/28067/26282.pdf
DOI:10.1016/j.aquaculture.2013.12.006
http://archimer.ifremer.fr/doc/00169/28067/

Éditeur(s) : HAL CCSD American Chemical Society
Résumé : International audience
2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), a carcinogenic heterocyclic aromatic amine formed in cooked meats, is metabolically activated to electrophilic intermediates that form covalent adducts with DNA and protein. We previously identified an adduct of PhIP formed at the Cys34 residue of human serum albumin following reaction of albumin with the genotoxic metabolite 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine (HONH-PhIP). The major adducted peptide recovered from a tryptic/chymotryptic digest was identified as the missed-cleavage peptide LQQC*[SO2PhIP]PFEDHVK, a [cysteine-S-yl-PhIP]-S-dioxide linked adduct. In this investigation, we have characterized the albumin adduction products of N-sulfooxy-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (N-sulfooxy-PhIP), which is thought to be a major genotoxic metabolite of PhIP formed in vivo. Targeted and data-dependent scanning methods showed that N-sulfooxy-PhIP adducted to the Cys34 of albumin in human plasma to form LQQC*[SO2PhIP]PFEDHVK at levels that were 8-10-fold greater than the adduct levels formed with N-(acetyloxy)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (N-acetoxy-PhIP) or HONH-PhIP. We also discovered that N-sulfooxy-PhIP forms an adduct at the sole tryptophan (Trp214) residue of albumin in the sequence AW*[PhIP]AVAR. However, stable adducts of PhIP with albumin were not detected in human hepatocytes. Instead, PhIP and 2-amino-1-methyl-6-(5-hydroxy)phenylimidazo[4,5-b]pyridine (5-HO-PhIP), a solvolysis product of the proposed nitrenium ion of PhIP, were recovered during the proteolysis, suggesting a labile sulfenamide linkage had formed between an N-oxidized intermediate of PhIP and Cys34 of albumin. A stable adduct was formed at the Tyr411 residue of albumin in hepatocytes and identified as a deaminated product of PhIP, Y*[desaminoPhIP]TK, where the 4-HO-tyrosine group bound to the C-2 imidazole atom of PhIP.
ISSN: 0893-228X

hal-01146777
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01146777
DOI : 10.1021/acs.chemrestox.5b00075
PUBMED : 25815793

Éditeur(s) : HAL CCSD Elsevier
Résumé : Recent evolutionary studies clearly indicate that evolution is mainly driven by changes in the complex mechanisms of gene regulation and not solely by polymorphism in protein-encoding genes themselves. After a short description of the cis-regulatory mechanism, we intend in this review to argue that by applying newly available technologies and by merging research areas such as evolutionary and developmental biology, population genetics, ecology and molecular cell biology it is now possible to study evolution in an integrative way. We contend that, by analysing the effects of promoter sequence variation on phenotypic diversity in natural populations, we will soon be able to break the barrier between the study of extant genetic variability and the study of major developmental changes. This will lead to an integrative view of evolution at different scales. Because of their sessile nature and their continuous development, plants must permanently regulate their gene expression to react to their environment, and can, therefore, be considered as a remarkable model for these types of studies. (C) 2010 Elsevier Ireland Ltd. All rights reserved.
ISSN: 0168-9452

hal-01032399
https://hal.archives-ouvertes.fr/hal-01032399
DOI : 10.1016/j.plantsci.2010.09.009

Éditeur(s) : HAL CCSD Wiley Open Access
Résumé : 11 pages
International audience
The loss of regulating agents such as parasites is among the most important changes in biotic interactions experienced by populations established in newly colonized areas. Under a relaxed parasite pressure, individuals investing less in costly immune mechanisms might experience a selective advantage and become successful colonizers as they re-allocate resources to other fitness-related traits. Accordingly, a refinement of the evolution of increased competitive ability (EICA) hypothesis proposed that immunity of invasive populations has evolved toward a reduced investment in innate immunity, the most costly component of immunity, and an increased humoral immunity that is less costly. Biogeographical approaches comparing populations between native and expansion ranges are particularly relevant in exploring this issue, but remain very scarce. We conducted a biogeographical comparison between populations of Spectacled Thrush (Turdus nudigenis) from the native area (South America) and from the expansion range (Caribbean islands). First, we compared haemosporidian prevalence and circulating haptoglobin (an acute-phase protein produced during inflammation). Second, we challenged captive birds from both ranges with Escherichia coli lipopolysaccharides (LPS) and measured postchallenge haptoglobin production and body mass change. Birds from the expansion range showed lower haemosporidian prevalence and lower levels of haptoglobin than birds from the native range. In addition, the inflammation elicited by LPS injection and its associated cost in terms of body mass loss were lower in birds from the expansion range than in birds from the native range. In accordance with the enemy release hypothesis, our results suggest that range expansion is associated with a reduced infection risk. Our study also supports the hypothesis that individuals from newly established populations have evolved mechanisms to dampen the inflammatory response and are in accordance with one prediction of the refined EICA hypothesis, proposed to understand biological invasions.
ISSN: 2045-7758

hal-01403291
https://hal.archives-ouvertes.fr/hal-01403291
DOI : 10.1002/ece3.2486

Éditeur(s) : HAL CCSD American Physiological Society
Résumé : International audience
Excessive or persistent inflammation and hepatocyte death are the key triggers of liver diseases. The poly(ADP-ribose) polymerase (PARP) proteins induce cell death and inflammation. Chemical inhibition of PARP activity protects against liver injury during concanavalin A (ConA)-induced hepatitis. In this mice model, ConA activates immune cells, which promote inflammation and induce hepatocyte death, mediated by the activated invariant natural killer T (iNKT) lymphocyte population. We analyzed immune cell populations in the liver and several lymphoid organs, such as the spleen, thymus, and bone marrow in Parp2-deficient mice to better define the role of PARP proteins in liver immunity and inflammation at steady state and during ConA-induced hepatitis. We show that 1) the genetic inactivation of Parp2, but not Parp1, protected mice from ConA hepatitis without deregulating cytokine expression and leucocyte recruitment; 2) cellularity was lower in the thymus, but not in spleen, liver, or bone marrow of Parp2-/- mice; 3) spleen and liver iNKT lymphocytes, as well as thymic T and NKT lymphocytes were reduced in Parp2 knockout mice. In conclusion, our results suggest that the defect of T-lymphocyte maturation in Parp2 knockout mice leads to a systemic reduction of iNKT cells, reducing hepatocyte death during ConA-mediated liver damage, thus protecting the mice from hepatitis.NEW & NOTEWORTHY The genetic inactivation of Parp2, but not Parp1, protects mice from concanavalin A hepatitis. Immune cell populations are lower in the thymus, but not in the spleen, liver, or bone marrow of Parp2-deficient mice compared with wild-type mice. Spleen and liver invariant natural killer T (NKT) lymphocytes, as well as thymic T and NKT lymphocytes, are reduced in Parp2-deficient mice
ISSN: 0193-1857

hal-01630440
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01630440
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01630440/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01630440/file/102513_1_merged_1499199857-1.pdf
DOI : 10.1152/ajpgi.00436.2016

Éditeur(s) : HAL CCSD Wiley-Blackwell
Résumé : International audience
Based on their small size and genomic properties, ultrasmall prokaryotic groups like the Candidate Phyla Radiation have been proposed as possible symbionts dependent on other bacteria or archaea. In this study, we use a bipartite graph analysis to examine patterns of sequence similarity between draft and complete genomes from ultrasmall bacteria and other complete prokaryotic genomes, assessing whether the former group might engage in significant gene transfer (or even endosymbioses) with other community members. Our results provide preliminary evidence for many lateral gene transfers with other prokaryotes, including members of the archaea, and report the presence of divergent, membrane-associated proteins among these ultrasmall taxa. In particular, these divergent genes were found in TM6 relatives of the intracellular parasite Babela massiliensis.
ISSN: 1462-2912

Droits : http://creativecommons.org/licenses/by/
hal-01480348
http://hal.upmc.fr/hal-01480348
http://hal.upmc.fr/hal-01480348/document
http://hal.upmc.fr/hal-01480348/file/emi13477.pdf
DOI : 10.1111/1462-2920.13477

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

Éditeur(s) : HAL CCSD Endocrine Society
Résumé : International audience
Sertoli cells (SCs), the only somatic cells within seminiferous tubules, associate intimately with developing germ cells. They not only provide physical and nutritional support but also secrete factors essential to the complex developmental processes of germ cell proliferation and differentiation. The SC transcriptome must therefore adapt rapidly during the different stages of spermatogenesis. We report comprehensive genome-wide expression profiles of pure populations of SCs isolated at 5 distinct stages of the first wave of mouse spermatogenesis, using RNA sequencing technology. We were able to reconstruct about 13 901 high-confidence, nonredundant coding and noncoding transcripts, characterized by complex alternative splicing patterns with more than 45% comprising novel isoforms of known genes. Interestingly, roughly one-fifth (2939) of these genes exhibited a dynamic expression profile reflecting the evolving role of SCs during the progression of spermatogenesis, with stage-specific expression of genes involved in biological processes such as cell cycle regulation, metabolism and energy production, retinoic acid synthesis, and blood-testis barrier biogenesis. Finally, regulatory network analysis identified the transcription factors endothelial PAS domain-containing protein 1 (EPAS1/Hif2α), aryl hydrocarbon receptor nuclear translocator (ARNT/Hif1β), and signal transducer and activator of transcription 1 (STAT1) as potential master regulators driving the SC transcriptional program. Our results highlight the plastic transcriptional landscape of SCs during the progression of spermatogenesis and provide valuable resources to better understand SC function and spermatogenesis and its related disorders, such as male infertility.
ISSN: 0888-8809

hal-01146846
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01146846
DOI : 10.1210/me.2014-1356
PUBMED : 25710594

Éditeur(s) : Co-tutelle Al Centro de Investigaciones Biológicas del Noroeste - Université de Bretagne occidentale
Résumé : The interest in probiotics for aquaculture is increasing rapidly. In the present work we evaluated the potential of some yeast to induce the larvae gut maturation in sea bass larvae Dicentrarchus labrax. The production of polyamines putrescine, spermidine and spermine was quantified in ten yeast strains using high performance liquid chromatography (HPLC). Afterwards, fluorescently labelled yeast adhesion to the intestine of sea bass larvae and the spotted sand bass (Paralabrax maculatofasciatus) was evaluated by fluorescent microscopy. Two of them, CBS 8339 strain (Debaryomyces hansenii) and X2180 strain (Saccharomyces cerevisiae) produce the polyamines of our interest. The same strains showed the best adhering capabilities to the sea bass and spotted sand bass intestine. Both strains were selected and introduced by pulverization into the diet of larvae. The larvae fed with CBS 8339 strain, increase survival in 8.3% but not in their weight. The secretion of trypsin and amylase as well as those from the brush border membrane: alkaline phosphatase, maltase and leucine aminopeptidase, are incresed at 27 days after hatching (dah). At 42 dah difference in the enzyme activity of the different groups does not exist, suggesting a maturation of the digestive system in early stages of larval development. A second experiment was carried out to know the effect of different levels of incorporation into the diet of CBS 8339 strain: 1.1% and 5.7%. The minimum level, 1.1% of yeast produce best growth and survival rates, as well as lower deformed larvae than those obtained by 5.7% and the control one. Both level of yeast inclusion favored the trypsin secretion and lipase activity at 26 dah. Whereas mRNA levels for trypsin and amylase were influenced by both yeast concentrations at 26 and 36 dah, which suggest a development of digestive capacity of larvae as a result of the presence of polyamines secreted by the yeast.
Les probiotiques ont été définis comme des préparations microbiennes vivantes utilisées comme additif alimentaire, et qui ont une action bénéfique sur l'animal hôte en améliorant la digestion et l'hygiène intestinale. L'application de tels traitements aux productions aquacoles suscite un intérêt qui s'accroît rapidement. Dans ce travail, nous avons étudié l'effet de souches de levures sur la maturation de l'intestin des larves du bar Dicentrarchus labrax. Ces souches produisent des polyamines qui sont impliquées dans la réplication et la différentiation cellulaire, ainsi que dans la synthèse des protéines et des acides nucléiques. Dix souches de levures ont été testées pour leur production de polyamines (putrescine, spermidine et spermine), et pour leur adhérence à la paroi intestinale du bar et la cabrilla (Paralabrax maculatofasciatus). La production des polyamines a été analysée en utilisant la chromatographie liquide à haute performance (HPLC). Les levures selectionées ont été marquées avec un composé fluorescent pour tester leur adhérence a l'intestin de larves de bar et de juvéniles de cabrilla. L'adhérence a pu ainsi être evaluée par observation microscopique en épifluorescence. Les souches CBS 8339 (Debaryomyces hansenii) et X2180 (Saccharomyces cerevisiae) ont été choisies pour leur forte production de spermidine et de spermine, respectivement, et pour leur capacité d'adhérence à l'intestin du bar et de la cabrilla. Les deux souches ont été ajoutées par pulvérisation au régime des larves. Nous avons observé une augmentation de 8.3% du taux de survie avec les larves alimentées avec la souche CBS 8339, mais le poids moyen de ces larves était inférieur à celui obtenu sans ajout de levure. Une augmentation de l'activité de la trypsine et de l'amylase, aussi bien que des activités enzymatiques dans la bordure en brosse, aminopeptidase, phosphatase alkaline et maltase, ont été observées dans les larves âgées de 27 jours alimentées avec la souche CBS 8339. Au jour 42, il n'existait plus de différence d'activité enzymatique entre les larves traitées ou non avec la levure. Ces résultats suggèrent que la maturation du système digestif a été accélérée chez les larves nourries avec la levure, la maturation étant achevée chez les larves de 42 jours, qu'elles aient reçu l'un ou l'autre régime. Une deuxième expérience a été effectuée pour étudier l'effet de différents niveaux d'incorporation dans le régime contenant 1,1% et 5,7% de levure CBS 8339. L'incorporation de 1,1% a induit une augmentation du poids moyen et du taux de survie des larves, ainsi qu'une réduction du taux de malformation de la colonne vertébrale, par rapport aux résultats obtenus avec 0 ou 5,7% de levure. Les deux niveaux d'incorporation de la levure ont favorisé l'activité de la trypsine et de la lipase au jour 26. Les niveaux d'ARNm codant pour la synthèse de la trypsine et de l'amylase ont été influencés par les deux doses de levure aux jours 26 et 36. Ces résultats suggèrent qu'une faible dose de levure améliore le développement et la capacité digestive des larves, et nous formulons l'hypothèse que cette efficacité est due à la production de polyamines in situ. Le rôle des polyamines est particulièrement important pendant les phases de forte croissance comme les stades larvaires, mais elles peuvent être toxiques à dose trop élevée. L'incorporation de levure dans l'aliment destiné aux larves semble donc une excellente méthode pour délivrer les polyamines chez l'animal hôte.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2002/these-103.pdf
http://archimer.ifremer.fr/doc/00000/103/

Éditeur(s) : HAL CCSD
Résumé : International audience
Many different proteins self-aggregate into insoluble fibrils growing apically by reversible addition of elementary building blocks. But beyond this common principle, the modalities of fibril formation are very disparate, with various intermediate forms which can be reshuffled by minor modifications of physico-chemical conditions or amino-acid sequences. To bypass this complexity, the multifaceted phenomenon of fibril formation is reduced here to its most elementary principles defined for a linear prototype of fibril. Selected generic features, including nucleation, elongation, and conformational recruitment, are modeled using minimalist hypotheses and tools, by separating equilibrium from kinetic aspects and in vitro from in vivo conditions. These reductionist approaches allow to bring out known and new rudiments, including the kinetic and equilibrium effects of nucleation, the dual influence of elongation on nucleation, the kinetic limitations on nucleation and fibril numbers, and the accumulation of complexes in vivo by rescue from degradation. Overlooked aspects of these processes are also pointed: the exponential distribution of fibril lengths can be recovered using various models because it is attributable to randomness only. It is also suggested that the same term "critical concentration" is used for different things, involved in either nucleation or elongation
Journal of Chemical Physics

hal-01263103
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01263103
DOI : 10.1063/1.4940149
PUBMED : 26801045

Éditeur(s) : HAL CCSD Cold Spring Harbor Laboratory Press
Résumé : International audience
Meiotic recombination is required for the segregation of homologous chromosomes and is essential for fertility. In most mammals, the DNA double-strand breaks (DSBs) that initiate meiotic recombination are directed to a subset of genomic loci (hot spots) by sequence-specific binding of the PRDM9 protein. Rapid evolution of the DNA-binding specificity of PRDM9 and gradual erosion of PRDM9-binding sites by gene conversion will alter the recombination landscape over time. To better understand the evolutionary turnover of recombination hot spots and its consequences, we mapped DSB hot spots in four major subspecies of Mus musculus with different Prdm9 alleles and in their F1 hybrids. We found that hot spot erosion governs the preferential usage of some Prdm9 alleles over others in hybrid mice and increases sequence diversity specifically at hot spots that become active in the hybrids. As crossovers are disfavored at such hot spots, we propose that sequence divergence generated by hot spot turnover may create an impediment for recombination in hybrids, potentially leading to reduced fertility and, eventually, speciation
ISSN: 0890-9369

hal-01281480
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01281480
DOI : 10.1101/gad.270009.115
PUBMED : 26833728