Pan-embryo cell dynamics of germlayer formation in zebrafish

Cell movements are coordinated across spatio-temporal scales to achieve precise positioning of organs during vertebrate gastrulation. In zebrafish, mechanisms governing such morphogenetic movements have so far only been studied within a local region or a single germlayer. Here, we present pan-embryo analyses of fate specification and dynamics of all three germlayers simultaneously within a gastrulating embryo, showing that cell movement characteristics are predominantly determined by its position within the embryo, independent of its germlayer identity. The spatially confined fate specification establishes a distinct distribution of cells in each germlayer during early gastrulation. The differences in the initial distribution are subsequently amplified by a unique global movement, which organizes the organ precursors along the embryonic body axis, giving rise to the blueprint of organ formation

INTEGRAL discovery of non-thermal hard X-ray emission from the Ophiuchus cluster

We present the results of deep observations of the Ophiuchus cluster of galaxies with INTEGRAL in the 3-80 keV band. We analyse 3 Ms of INTEGRAL data on the Ophiuchus cluster with the IBIS/ISGRI hard X-ray imager and the JEM-X X-ray monitor. In the X-ray band using JEM-X, we show that the source is extended, and that the morphology is compatible with the results found by previous missions. Above 20 keV, we show that the size of the source is slightly larger than the PSF of the instrument, and is consistent with the soft X-ray morphology found with JEM-X and ASCA. Thanks to the constraints on the temperature provided by JEM-X, we show that the spectrum of the cluster is not well fitted by a single-temperature thermal Bremsstrahlung model, and that another spectral component is needed to explain the high energy data. We detect the high energy tail with a higher detection significance (6.4 sigma) than the BeppoSAX claim (2 sigma). Because of the imaging capabilities of JEM-X and ISGRI, we are able to exclude the possibility that the excess emission comes from very hot regions or absorbed AGN, which proves that the excess emission is indeed of non-thermal origin. Using the available radio data together with the non-thermal hard X-ray flux, we estimate a magnetic field B ~ 0.1-0.2 mu G.Comment: 8 pages, 9 figures, accepted by A&

GAA repeat expansion mutation mouse models of Friedreich ataxia exhibit oxidative stress leading to progressive neuronal and cardiac pathology

Friedreich ataxia (FRDA) is a neurodegenerative disorder caused by an unstable GAA repeat expansion mutation within intron 1 of the FXN gene. However, the origins of the GAA repeat expansion, its unstable dynamics within different cells and tissues, and its effects on frataxin expression are not yet completely understood. Therefore, we have chosen to generate representative FRDA mouse models by using the human FXN GAA repeat expansion itself as the genetically modified mutation. We have previously reported the establishment of two lines of human FXN YAC transgenic mice that contain unstable GAA repeat expansions within the appropriate genomic context. We now describe the generation of FRDA mouse models by crossbreeding of both lines of human FXN YAC transgenic mice with heterozygous Fxn knockout mice. The resultant FRDA mice that express only human-derived frataxin show comparatively reduced levels of frataxin mRNA and protein expression, decreased aconitase activity, and oxidative stress, leading to progressive neurodegenerative and cardiac pathological phenotypes. Coordination deficits are present, as measured by accelerating rotarod analysis, together with a progressive decrease in locomotor activity and increase in weight. Large vacuoles are detected within neurons of the dorsal root ganglia (DRG), predominantly within the lumbar regions in 6-month-old mice, but spreading to the cervical regions after 1 year of age. Secondary demyelination of large axons is also detected within the lumbar roots of older mice. Lipofuscin deposition is increased in both DRG neurons and cardiomyocytes, and iron deposition is detected in cardiomyocytes after 1 year of age. These mice represent the first GAA repeat expansion-based FRDA mouse models that exhibit progressive FRDA-like pathology and thus will be of use in testing potential therapeutic strategies, particularly GAA repeat-based strategies. © 2006 Elsevier Inc. All rights reserved

Architecture of the fungal nuclear pore inner ring complex

The nuclear pore complex (NPC) constitutes the sole gateway for bidirectional nucleocytoplasmic transport. We present the reconstitution and interdisciplinary analyses of the ~425-kDa inner ring complex (IRC), which forms the central transport channel and diffusion barrier of the NPC, revealing its interaction network and equimolar stoichiometry. The Nsp1•Nup49•Nup57 channel nucleoporin hetero-trimer (CNT) attaches to the IRC solely through the adaptor nucleoporin Nic96. The CNT•Nic96 structure reveals that Nic96 functions as an assembly sensor that recognizes the three dimensional architecture of the CNT, thereby mediating the incorporation of a defined CNT state into the NPC. We propose that the IRC adopts a relatively rigid scaffold that recruits the CNT to primarily form the diffusion barrier of the NPC, rather than enabling channel dilation

Radio Halos From Simulations And Hadronic Models II: The Scaling Relations of Radio Halos

We use results from a constrained, cosmological MHD simulation of the Local Universe to predict radio halos and their evolution for a volume limited set of galaxy clusters and compare to current observations. The simulated magnetic field inside the clusters is a result of turbulent amplification within them, with the magnetic seed originating from star-burst driven, galactic outflows. We evaluate three models, where we choose different normalizations for the Cosmic Ray proton population within clusters. Similar to our previous analysis of the Coma cluster (Donnert et al. 2010), the radial profile and the morphological properties of observed radio halos can not be reproduced, even with a radially increasing energy fraction within the cosmic ray proton population. Scaling relations between X-ray luminosity and radio power can be reproduced by all models, however all models fail in the prediction of clusters with no radio emission. Also the evolutionary tracks of our largest clusters in all models fail to reproduce the observed bi-modality in radio luminosity. This provides additional evidence that the framework of hadronic, secondary models is disfavored to reproduce the large scale diffuse radio emission of galaxy clusters. We also provide predictions for the unavoidable emission of $\gamma$-rays from the hadronic models for the full cluster set. None of such secondary models is yet excluded by the observed limits in $\gamma$-ray emission, emphasizing that large scale diffuse radio emission is a powerful tool to constrain the amount of cosmic ray protons in galaxy clusters

Variable Expression of Cre Recombinase Transgenes Precludes Reliable Prediction of Tissue-Specific Gene Disruption by Tail-Biopsy Genotyping

The Cre/loxP-system has become the system of choice for the generation of conditional so-called knockout mouse strains, i.e. the tissue-specific disruption of expression of a certain target gene. We here report the loss of expression of Cre recombinase in a transgenic mouse strain with increasing number of generations. This eventually led to the complete abrogation of gene expression of the inserted Cre cDNA while still being detectable at the genomic level. Conversely, loss of Cre expression caused an incomplete or even complete lack of disruption for the protein under investigation. As Cre expression in the tissue of interest in most cases cannot be addressed in vivo during the course of a study, our findings implicate the possibility that individual tail-biopsy genotypes may not necessarily indicate the presence or absence of gene disruption. This indicates that sustained post hoc analyses in regards to efficacy of disruption for every single study group member may be required

Radio Halos From Simulations And Hadronic Models I: The Coma cluster

We use the results from a constrained, cosmological MHD simulation of the Local Universe to predict the radio halo and the gamma-ray flux from the Coma cluster and compare it to current observations. The simulated magnetic field within the Coma cluster is the result of turbulent amplification of the magnetic field during build-up of the cluster. The magnetic seed field originates from star-burst driven, galactic outflows. The synchrotron emission is calculated assuming a hadronic model. We follow four approaches with different distributions for the cosmic-ray proton (CRp) population within galaxy clusters. The radial profile the radio halo can only be reproduced with a radially increasing energy fraction within the cosmic ray proton population, reaching $>$100% of the thermal energy content at $\approx$ 1Mpc, e.g. the edge of the radio emitting region. Additionally the spectral steepening of the observed radio halo in Coma cannot be reproduced, even when accounting for the negative flux from the thermal SZ effect at high frequencies. Therefore the hadronic models are disfavored from present analysis. The emission of $\gamma$-rays expected from our simulated coma is still below the current observational limits (by a factor of $\sim$6) but would be detectable in the near future.Comment: Submitted to MNRAS, 5pages, 3 figures, 1 tabl

Clusters of galaxies : observational properties of the diffuse radio emission

Clusters of galaxies, as the largest virialized systems in the Universe, are ideal laboratories to study the formation and evolution of cosmic structures...(abridged)... Most of the detailed knowledge of galaxy clusters has been obtained in recent years from the study of ICM through X-ray Astronomy. At the same time, radio observations have proved that the ICM is mixed with non-thermal components, i.e. highly relativistic particles and large-scale magnetic fields, detected through their synchrotron emission. The knowledge of the properties of these non-thermal ICM components has increased significantly, owing to sensitive radio images and to the development of theoretical models. Diffuse synchrotron radio emission in the central and peripheral cluster regions has been found in many clusters. Moreover large-scale magnetic fields appear to be present in all galaxy clusters, as derived from Rotation Measure (RM) studies. Non-thermal components are linked to the cluster X-ray properties, and to the cluster evolutionary stage, and are crucial for a comprehensive physical description of the intracluster medium. They play an important role in the cluster formation and evolution. We review here the observational properties of diffuse non-thermal sources detected in galaxy clusters: halos, relics and mini-halos. We discuss their classification and properties. We report published results up to date and obtain and discuss statistical properties. We present the properties of large-scale magnetic fields in clusters and in even larger structures: filaments connecting galaxy clusters. We summarize the current models of the origin of these cluster components, and outline the improvements that are expected in this area from future developments thanks to the new generation of radio telescopes.Comment: Accepted for the publication in The Astronomy and Astrophysics Review. 58 pages, 26 figure

Architecture of the symmetric core of the nuclear pore

INTRODUCTION: The nuclear pore complex (NPC) is the primary gateway for the transport of macromolecules between the nucleus and cytoplasm, serving as both a critical mediator and regulator of gene expression. NPCs are very large (~120 MDa) macromolecular machines embedded in the nuclear envelope, each containing ~1000 protein subunits, termed nucleoporins. Despite substantial progress in visualizing the overall shape of the NPC by means of cryoelectron tomography (cryo-ET) and in determining atomic-resolution crystal structures of nucleoporins, the molecular architecture of the assembled NPC has thus far remained poorly understood, hindering the design of mechanistic studies that could investigate its many roles in cell biology. RATIONALE: Existing cryo-ET reconstructions of the NPC are too low in resolution to allow for de novo structure determination of the NPC or unbiased docking of nucleoporin fragment crystal structures. We sought to bridge this resolution gap by first defining the interaction network of the NPC, focusing on the evolutionarily conserved symmetric core. We developed protocols to reconstitute NPC protomers from purified recombinant proteins, which enabled the generation of a high-resolution biochemical interaction map of the NPC symmetric core. We next determined high-resolution crystal structures of key nucleoporin interactions, providing spatial restraints for their relative orientation. By superposing crystal structures that overlapped in sequence, we generated accurate full-length structures of the large scaffold nucleoporins. Lastly, we used sequential unbiased searches, supported by the biochemical data, to place the nucleoporin crystal structures into a previously determined cryo-ET reconstruction of the intact human NPC, thus generating a composite structure of the entire NPC symmetric core. RESULTS: Our analysis revealed that the inner and outer rings of the NPC use disparate mechanisms of interaction. Whereas the structured coat nucleoporins of the outer ring form extensive surface contacts, the scaffold proteins of the inner ring are bridged by flexible sequences in linker nucleoporins. Our composite structure revealed a defined spoke architecture in which each of the eight spokes spans the nuclear envelope, with limited cross-spoke interactions. Most nucleoporins are present in 32 copies, with the exceptions of Nup170 and Nup188, which are present in 48 and 16 copies, respectively. Lastly, we observed the arrangement of the channel nucleoporins, which orient their N termini into two 16-membered rings, thus ensuring that their N-terminal FG repeats project evenly into the central transport channel. CONCLUSION: Our composite structure of the NPC symmetric core can be used as a platform for the rational design of experiments to investigate NPC structure and function. Each nucleoporin occupies multiple distinct biochemical environments, explaining how such a large macromolecular complex can be assembled from a relatively small number of genes. Our integrated, bottom-up approach provides a paradigm for the biochemical and structural characterization of similarly large biological mega-assemblies