Krüppel-like transcription factors and control of pluripotency

Recent papers have demonstrated a role for Krüppel-like transcription factors 2, 4 and 5 in the control of mouse embryonic stem cell pluripotency. However, it is not clear whether each factor has a unique role or whether they are functionally redundant. A paper by Parisi and colleagues in BMC Biology now sheds light on the mechanism by which Klf5 regulates pluripotency

Etude dynamique et structurale de biomolécules par microscopie à force atomique HS-AFM (application à une petite protéine de choc thermique sHsp)

La microscopie à force atomique (AFM) permet de visualiser la topographie d échantillons organiqueset inorganiques à l échelle atomique. Les innovations les plus récentes offrent désormais la possibilitéd accéder aux propriétés nano-mécaniques des échantillons (élasticité, adhésion ). Son panel defonctionnalités permet de pallier aux besoins des nanotechnologies, tant dans les domaines de laphysique, de la chimie que de la biologie.Cependant, les besoins nécessaires à la compréhension des processus biologiques imposent aumicroscope à force atomique des vitesses d acquisitions rapides, inférieures à la seconde par image. Leséquipements classiques n offrent pas cette possibilité. C est pour s affranchir de ce verrou technologique,pour l étude dynamique, qu un prototype de microscope à force atomique à haute-vitesse a étédéveloppé (HS-AFM) en partenariat avec l équipe du Professeur T. Ando à l Université de Kanazawa(Japon). Il permet d atteindre des vitesses de balayage identiques aux vitesses vidéos : 25-50 images/s, enmilieu liquide. Le dispositif est en perpétuelle amélioration : nouvelle boucle d asservissement, domainesde balayage augmentés. La haute résolution est, quant à elle, assurée par des leviers miniaturisés munisde sur-pointes en carbone. Parallèlement à l innovation du microscope en lui-même, des modulescomplémentaires ont été développés : module pousse seringue et module chauffant.Le potentiel de ce prototype, développé dans le cadre d un programme ANR PNANO 2008 HSnanobio-Imaging, a été montré via l étude d une petite protéine de choc thermique : la protéine sHspLo18. Cette protéine, issue de la bactérie lactique Oenococcus oeni, offrait la possibilité d étudier deschangements de degrés d oligomérisation en fonction du pH, ainsi que le rôle chaperon et lipochaperonen cas de stress environnemental d autres complexes biologiques. L utilisation des techniques demicroscopie couplée à des études biochimiques sur ce modèle protéique a permis d appréhender l effetdes surfaces sur l adsorption et la dynamique des complexes biologiques. L interaction protéine surfacea pu être approchée et s avère utile au développement des capteurs à protéinesThe atomic force microscopy (AFM) gives access to the topography of organic and inorganic samplesat the atomic scale. The latest innovations offer the possiblity to understand the sample nano-mechanicalproperties (elasticity, adhesion...). Its feature set allows overcoming the demands of nanotechnology,both in the fields of physics, chemistry and biology.However, understanding biological processes require faster acquisitions for the atomic forcemicroscopy, less than a second per frame. As conventional equipment does not offer the possibility toovercome the constraint of time for dynamical studies, a prototype of high-speed atomic forcemicroscope (HS-AFM) was developed in partnership with Professor T. Ando group of Kanazawa University(Japan). It can reach scanning video speed: 25-50 frames/s in a liquid medium. The device is beingconstantly improved: new feedback control, larger scanning sizes. The resolution is provided byminiaturized cantilevers with carbon EBD-tips. In parallel to innovative modules on the microscope, addonshave been developed: syringe pump and heating modules.The potential of the prototype, developed within the framework of the program ANR PNANO 2008HS-nanobio-Imaging, has been shown through the study of a small heat shock protein: the protein sHspLo18. This protein, from the lactic acid bacterium Oenococcus oeni, offered the possibility of a variouschanges of oligomerization degrees according to the pH, and also the chaperone and lipochaperon activityof protein under the influence of an environmental stress. The use of these techniques of microscopiescoupled with biochemical studies on this proteic model allowed to dread the effect of surfaces on theadsorption and the dynamics of biological complexes. The interaction protein surface coulb be toapprehend and proves to be useful for the development of protein sensors developed in the laboratoryDIJON-BU Doc.électronique (212319901) / SudocSudocFranceF

Epigenetic status of H19/IGF2 and SNRPN imprinted genes in aborted and successfully derived embryonic stem cell lines in non-human primates

AbstractThe imprinted genes of primate embryonic stem cells (ESCs) often show altered DNA methylation. It is unknown whether these alterations emerge while deriving the ESCs. Here we studied the methylation patterns of two differentially methylated regions (DMRs), SNRPN and H19/IGF2 DMRs, during the derivation of monkey ESCs. We show that the SNRPN DMR is characteristically methylated at maternal alleles, whereas the H19/IGF2 DMR is globally highly methylated, with unusual methylation on the maternal alleles. These methylation patterns remain stable from the early stages of ESC derivation to late passages of monkey ESCs and following differentiation. Importantly, the methylation status of H19/IGF2 DMR and the expression levels of IGF2, H19, and DNMT3B mRNAs in early embryo-derived cells were correlated with their capacity to generate genuine ESC lines. Thus, we propose that these markers could be useful to predict the outcomes of establishing an ESC line in primates

High-resolution characterization of the diffusion of light chemical elements in metallic components by scanning microwave microscopy

International audienceAn original sub-surface, high spatial resolution tomographic technique based on scanning microwave microscopy (SMM) is used to visualize in-depth materials with different chemical compositions. A significant phase difference in SMM between aluminum and chromium buried patterns has been observed. Moreover this technique was used to characterize a solid solution of a light chemical element (oxygen) in a metal lattice (zirconium). The large solubility of the oxygen in zirconium leads to modifications of the properties of the solid solution that can be measured by the phase shift signal in the SMM technique. The signal obtained in cross-section of an oxidized Zr sample shows the excellent agreement between phase shift profiles measured at different depths. Such a profile can reveal the length of diffusion of the oxygen in zirconium under the surface. The comparison with the oxygen concentration measured by nuclear reaction analysis shows excellent agreement in terms of length of diffusion and spatial distribution of the oxygen. A rapid calibration shows a linear dependence between the phase shift and the oxygen concentration. The SMM method opens up new possibilities for indirect measurements of the oxygen concentration dissolved in the metal lattic

External controls on the distribution, fabrics and mineralization of modern microbial mats in a coastal hypersaline lagoon, Cayo Coco (Cuba).

45 pagesInternational audienceActive, carbonate-mineralizing microbial mats flourish in a tropical, highly evaporative, marine-fed lagoonal network to the south of Cayo Coco Island (Cuba). Hypersaline conditions support the development of a complex sedimentary microbial ecosystem with diverse morphologies, a variable intensity of mineralization and a potential for preservation. In this study, the role of intrinsic (i.e. microbial) and extrinsic (i.e. physicochemical) controls on microbial mat development, mineralization and preservation was investigated. The network consists of lagoons, forming in the interdune depressions of a Pleistocene aeolian substratum; they developed due to a progressive increase in sea-level since the Holocene. The hydrological budget in the Cayo Coco lagoonal network changes from west to east, increasing the salinity. This change progressively excludes grazers and increases the saturation index of carbonate minerals, favouring the development and mineralization of microbial mats in the easternmost lagoons. Detailed mapping of the easternmost lagoon shows four zones with different flooding regimes. The microbial activity in the mats was recorded using light–dark shifts in conjunction with microelectrode O2 and HS− profiles. High rates of O2 production and consumption, in addition to substantial amounts of exopolymeric substances, are indicative of a potentially strong intrinsic control on mineralization. Seasonal, climate-driven water fluctuations are key for mat development, mineralization, morphology and distribution. Microbial mats show no mineralization in the permanently submersed zone, and moderate mineralization in zones with alternating immersion and exposure. It is suggested that mineralization is also driven by water-level fluctuations and evaporation. Mineralized mats are laminated and consist of alternating trapping and binding of grains and microbially induced magnesium calcite and dolomite precipitation. The macrofabrics of the mats evolve from early colonizing Flat mats to complex Cerebroid or Terrace structures. The macrofabrics are influenced by the hydrodynamic regime: wind-driven waves inducing relief terraces in windward areas and flat morphologies on the leeward side of the lagoon. Other external drivers include: (i) storm events that either promote (for example, by bioclasts covering) or prevent (for example, by causing erosion) microbial mat preservation; and (ii) subsurface degassing, through mangrove roots and desiccation cracks covered by Flat mats (i.e. forming Hemispheroids and Cerebroidal structures). These findings provide in-depth insights into understanding fossil microbialite morphologies that formed in lagoonal settings

Advances in quantitative nanoscale subsurface imaging by mode-synthesizing atomic force microscopy

This paper reports on advances toward quantitative non-destructive nanoscale subsurface investigation of a nanofabricated sample based on mode synthesizing atomic force microscopy with heterodyne detection, addressing the need to correlate the role of actuation frequencies of the probe f(p) and the sample f(s) with depth resolution for 3D tomography reconstruction. Here, by developing a simple model and validating the approach experimentally through the study of the nanofabricated calibration depth samples consisting of buried metallic patterns, we demonstrate avenues for quantitative nanoscale subsurface imaging. Our findings enable the reconstruction of the sample depth profile and allow high fidelity resolution of the buried nanostructures. Non-destructive quantitative nanoscale subsurface imaging offers great promise in the study of the structures and properties of complex systems at the nanoscale

A short G1 phase is an intrinsic determinant of naïve embryonic stem cell pluripotency

AbstractA short G1 phase is a characteristic feature of mouse embryonic stem cells (ESCs). To determine if there is a causal relationship between G1 phase restriction and pluripotency, we made use of the Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) reporter system to FACS-sort ESCs in the different cell cycle phases. Hence, the G1 phase cells appeared to be more susceptible to differentiation, particularly when ESCs self-renewed in the naïve state of pluripotency. Transitions from ground to naïve, then from naïve to primed states of pluripotency were associated with increased durations of the G1 phase, and cyclin E-mediated alteration of the G1/S transition altered the balance between self-renewal and differentiation. LIF withdrawal resulted in a lengthening of the G1 phase in naïve ESCs, which occurred prior to the appearance of early lineage-specific markers, and could be reversed upon LIF supplementation. We concluded that the short G1 phase observed in murine ESCs was a determinant of naïve pluripotency and was partially under the control of LIF signaling

Late glacial to deglacial variation of coralgal assemblages in the Great Barrier Reef, Australia

Integrated Ocean Drilling Program (IODP) Expedition 325 cored submerged reefs along the shelf edge of the Great Barrier Reef (GBR) to study sea-level and environmental changes and their impacts on reef communities and reef growth since the Last Glacial Maximum (LGM). Previous work defined five reef sequences (Reef 1–5) that span the last 30,000 years. Here we examined the variation in coralgal assemblages and their paleoenvironmental settings in late glacial to deglacial sequences from 23 holes cored seaward of the modern GBR in water depths from 46 to 131 m along four transects at three localities: Hydrographers Passage (HYD-01C and HYD-02A), Noggin Pass (NOG–01B), and Ribbon Reef (RIB-02A). We identified three coralline algal assemblages and eight coral assemblages indicating a broad range of reef settings from the shallow reef crest (0–5 m) to the deep forereef slope (>20 m). We document in detail for the first time the distribution and composition of reef communities that grew in the GBR during the LGM from 22,000–19,000 years ago. They included coral taxa that are major reef builders today: Isopora, Acropora gr. humilis, Dipsastraea gr. pallida, Porites, and Montipora. Prior to the fall in sea level to the maximum extent of the LGM, late glacial reef communities developed more proximally (landward) to the modern GBR along the shelf edge. Their distribution and composition reflect influences of the older Pleistocene basement depth and possible terrigenous sediment inputs. Post-LGM deglacial reef growth was vigorous in proximal sites and characterized by the accretion of a very shallow high-energy coralgal assemblage composed of medium to robustly branching Acropora, including A. gr. humilis, and thick algal crusts of Porolithon gr. onkodes associated with vermetid gastropods. More distally, reef growth was variably impacted by terrigenous input following deglacial reflooding of antecedent reef terraces. The coralgal succession and sedimentary facies in Noggin Pass indicate that an early drowning trend was linked to increased turbidity that was likely controlled by shelf morphology (narrow shelf, steep slope) and/or proximity to a paleo-river mouth. The deglacial succession in Ribbon Reef lacks typical shallow-water indicators, which may reflect influences of the particularly steep slope of the northern GBR shelf edge on reef zonation. A major sea-level jump at the onset of the Younger Dryas displaced reef habitats further upslope, forming a barrier reef system mainly composed of robustly branching acroporids distinct from the more distal sites. Our results highlight the importance of sedimentation and shelf morphology in addition to relative sea-level changes in controlling variations in reef community over centennial to millennial timescales. © 2019 Elsevier B.V.Australian Research Council-DP109400

Microbial and diagenetic steps leading to the mineralisation of Great Salt Lake microbialites.

12 pagesInternational audienceMicrobialites are widespread in modern and fossil hypersaline environments, where they provide a unique sedimentary archive. Authigenic mineral precipitation in modern microbialites results from a complex interplay between microbial metabolisms, organic matrices and environmental parameters. Here, we combined mineralogical and microscopic analyses with measurements of metabolic activity in order to characterise the mineralisation of microbial mats forming microbialites in the Great Salt Lake (Utah, USA). Our results show that the mineralisation process takes place in three steps progressing along geochemical gradients produced through microbial activity. First, a poorly crystallized Mg-Si phase precipitates on alveolar extracellular organic matrix due to a rise of the pH in the zone of active oxygenic photosynthesis. Second, aragonite patches nucleate in close proximity to sulfate reduction hotspots, as a result of the degradation of cyanobacteria and extracellular organic matrix mediated by, among others, sulfate reducing bacteria. A final step consists of partial replacement of aragonite by dolomite, possibly in neutral to slightly acidic porewater. This might occur due to dissolution-precipitation reactions when the most recalcitrant part of the organic matrix is degraded. The mineralisation pathways proposed here provide pivotal insight for the interpretation of microbial processes in past hypersaline environments