Distinct mechanisms regulate Cdx2 expression in the blastocyst and in trophoblast stem cells

The first intercellular differences during mammalian embryogenesis arise in the blastocyst, producing the inner cell mass and the trophectoderm. The trophectoderm is the first extraembryonic tissue and does not contribute to the embryo proper, its differentiation instead forming tissues that sustain embryonic development. Crucial roles in extraembryonic differentiation have been identified for certain transcription factors, but a comprehensive picture of the regulation of this early specification is still lacking. Here, we investigated whether the regulatory mechanisms involved in Cdx2 expression in the blastocyst are also utilized in the postimplantation embryo. We analyzed an enhancer that is regulated through Hippo and Notch in the blastocyst trophectoderm, unexpectedly finding that it is inactive in the extraembryonic structures at postimplantation stages. Further analysis identified other Cdx2 regulatory elements including a stem-cell specific regulatory sequence and an element that drives reporter expression in the trophectoderm, a subset of cells in the extraembryonic region of the postimplantation embryo and in trophoblast stem cells. The cross-comparison in this study of cis-regulatory elements employed in the blastocyst, stem cell populations and the postimplantation embryo provides new insights into early mammalian development and suggests a two-step mechanism in Cdx2 regulation.We thank Barbara Pernaute for comments and suggestions; members of the Manzanares lab for comments, technical help and support; Ian Chambers and Austin Smith for the ZHBTc4 ES cell line; Tristan Rodriguez for the B1-TS cell line; Luis Miguel Criado and the CNIC Transgenesis Unit for TS cell morulae injections and support; Roisin Doohan for help with sections; and Simon Bartlett (CNIC) for English editing. This study was funded by grants from the Ministerio de Economia y Competitividad (grant BFU2011-23083 and BFU2014-54608-P to MM; FPU Doctoral Fellowship to TR; FPI-SO Doctoral Fellowship to SM; Severo Ochoa Center of Excellence award SEV-2015-0505 to CNIC), Comunidad Autonoma de Madrid (grant CELLDD-CM to MM), Canadian Institute of Health Research (JR), Imperial College (VA), and the MRC and Genesis Research Trust (AH). The CNIC is supported by the Spanish Ministerio de Economia y Competitividad and the Pro-CNIC Foundation.S

Nervous System Regionalization Entails Axial Allocation before Neural Differentiation

Neural induction in vertebrates generates a CNS that extends the rostral-caudal length of the body. The prevailing view is that neural cells are initially induced with anterior (forebrain) identity; caudalizing signals then convert a proportion to posterior fates (spinal cord). To test this model, we used chromatin accessibility to define how cells adopt region-specific neural fates. Together with genetic and biochemical perturbations, this identified a developmental time window in which genome-wide chromatin-remodeling events preconfigure epiblast cells for neural induction. Contrary to the established model, this revealed that cells commit to a regional identity before acquiring neural identity. This "primary regionalization" allocates cells to anterior or posterior regions of the nervous system, explaining how cranial and spinal neurons are generated at appropriate axial positions. These findings prompt a revision to models of neural induction and support the proposed dual evolutionary origin of the vertebrate CNS

Study of doubly strange systems using stored antiprotons

Bound nuclear systems with two units of strangeness are still poorly known despite their importance for many strong interaction phenomena. Stored antiprotons beams in the GeV range represent an unparalleled factory for various hyperon-antihyperon pairs. Their outstanding large production probability in antiproton collisions will open the floodgates for a series of new studies of systems which contain two or even more units of strangeness at the P‾ANDA experiment at FAIR. For the first time, high resolution γ-spectroscopy of doubly strange ΛΛ-hypernuclei will be performed, thus complementing measurements of ground state decays of ΛΛ-hypernuclei at J-PARC or possible decays of particle unstable hypernuclei in heavy ion reactions. High resolution spectroscopy of multistrange Ξ−-atoms will be feasible and even the production of Ω−-atoms will be within reach. The latter might open the door to the |S|=3 world in strangeness nuclear physics, by the study of the hadronic Ω−-nucleus interaction. For the first time it will be possible to study the behavior of Ξ‾+ in nuclear systems under well controlled conditions

Ab initio and experimental studies on the hetero-Diels-Alder and cheletropic additions of sulfur dioxide to (E)-1-methoxybutadiene: A mechanism involving three molecules of SO2

Kinetics on the cheletropic addition of sulfur dioxide to (E)-1-methoxybutadiene (1) to give the corresponding sulfolene 2 (2-methoxy-2,5-dihydrothiophene-1,1-dioxide) gave the rate law d[2]/dt = k[1] [SO2](x) with x = 2.6 +/- 0.2 at 198 K. Under these conditions, no sultine 3 [(2RS,6RS)-6-methoxy-3,6-dihydro-1,2-oxathiin-2-oxide] resulting from a hetero-Diels-Alder addition was observed, and the cheletropic elimination 2 --> 1 + SO2 did not occur. Ab initio and DFT quantum calculations confirmed that the cheletropic addition 1 + SO2 --> 2 follows two parallel mechanisms, one involving two molecules of SO2 and the transition structure with DeltaG(double dagger) = 18.2 +/- 0.2 kcal/mol at 198 K (exptl); 22.5-22.7 kcal/mol [B3LYP/6-31G(d,p)], the other one involving three molecules of SO2 with DeltaG(double dagger) = 18.9 +/- 0.1 kcal/mol at 198 K (exptl); 19.7 kcal/mol [B3LYP/6-31G(d,p)]. The mechanism involving only one molecule of SO2 in the transition structure requires a higher activation energy, DeltaG(double dagger) = 25.2 kcal/mol [B3LYP/6-31G(d,p)]. Comparison of the geometries and energetics of the structures involved into the 1 + SO2 --> 2, 3 and 1 + 2SO(2) --> 2, 3 + SO2 reactions obtained by ab initio and DFT methods suggest that the latter calculation techniques can be used to study the cycloadditions of sulfur dioxide. The calculations predict that the hetero-Diels-Alder addition 1 + SO2 --> 3 also prefers a mechanism in which three molecules of SO2 are involved in the cycloaddition transition structure. At 198 K and in SO2 solutions, the entropy cost (TDeltaS(double dagger)) is overcompensated by the specific solvation by SO2 in the transition structures of both the cheletropic and hetero-Diels-Alder reactions of (E)-1-methoxybutadiene with SO2

Ab initio and experimental studies on the hetero-Diels-Alder and cheletropic additions of sulfur dioxide to (E)-1-methoxybutadiene: A mechanism involving three molecules of SO2

Kinetics on the cheletropic addition of sulfur dioxide to (E)-1-methoxybutadiene (1) to give the corresponding sulfolene 2 (2-methoxy-2,5-dihydrothiophene-1,1-dioxide) gave the rate law d[2]/dt = k[1] [SO2](x) with x = 2.6 +/- 0.2 at 198 K. Under these conditions, no sultine 3 [(2RS,6RS)-6-methoxy-3,6-dihydro-1,2-oxathiin-2-oxide] resulting from a hetero-Diels-Alder addition was observed, and the cheletropic elimination 2 --> 1 + SO2 did not occur. Ab initio and DFT quantum calculations confirmed that the cheletropic addition 1 + SO2 --> 2 follows two parallel mechanisms, one involving two molecules of SO2 and the transition structure with DeltaG(double dagger) = 18.2 +/- 0.2 kcal/mol at 198 K (exptl); 22.5-22.7 kcal/mol [B3LYP/6-31G(d,p)], the other one involving three molecules of SO2 with DeltaG(double dagger) = 18.9 +/- 0.1 kcal/mol at 198 K (exptl); 19.7 kcal/mol [B3LYP/6-31G(d,p)]. The mechanism involving only one molecule of SO2 in the transition structure requires a higher activation energy, DeltaG(double dagger) = 25.2 kcal/mol [B3LYP/6-31G(d,p)]. Comparison of the geometries and energetics of the structures involved into the 1 + SO2 --> 2, 3 and 1 + 2SO(2) --> 2, 3 + SO2 reactions obtained by ab initio and DFT methods suggest that the latter calculation techniques can be used to study the cycloadditions of sulfur dioxide. The calculations predict that the hetero-Diels-Alder addition 1 + SO2 --> 3 also prefers a mechanism in which three molecules of SO2 are involved in the cycloaddition transition structure. At 198 K and in SO2 solutions, the entropy cost (TDeltaS(double dagger)) is overcompensated by the specific solvation by SO2 in the transition structures of both the cheletropic and hetero-Diels-Alder reactions of (E)-1-methoxybutadiene with SO2

Regionalization of the nervous system requires axial allocation prior to neural lineage commitment

Neural induction in vertebrates generates a central nervous system that extends the rostral-caudal length of the body. The prevailing view is that neural cells are initially induced with anterior (forebrain) identity, with caudalising signals then converting a proportion to posterior fates (spinal cord). To test this model, we used chromatin accessibility assays to define how cells adopt region-specific neural fates. Together with genetic and biochemical perturbations this identified a developmental time window in which genome-wide chromatin remodeling events preconfigure epiblast cells for neural induction. Contrary to the established model, this revealed that cells commit to a regional identity before acquiring neural identity. This “primary regionalization” allocates cells to anterior or posterior regions of the nervous system, explaining how cranial and spinal neurons are generated at appropriate axial positions. These findings prompt a revision to models of neural induction and support the proposed dual evolutionary origin of the vertebrate central nervous system

On the Molecular Structure of Uranium Dicarbide: T-Shape versus Linear Isomers

A theoretical study of the molecular structure of uranium dicarbide has been carried out employing DFT, coupled cluster, and multiconfigurational methods. A triangular species, corresponding to a 5A 2 electronic state, has been found to be the most stable UC 2 species. A triplet linear CUC species, which has been observed in recent infrared spectroscopy experiments, lies much higher in energy. A topological analysis of the electronic density has also been carried out. The triangular species is shown to be in fact a T-shape structure with a U-C interaction which can be considered to be a closed-shell interaction.Fil: Zalazar, Maria Fernanda. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura. Departamento de Química. Laboratorio de Estructura Molecular y Propiedades; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Rayon, Victor M.. Universidad de Valladolid. Facultad de Ciencias; EspañaFil: Largo, Antonio. Universidad de Valladolid. Facultad de Ciencias; Españ