Regional requirements for Dishevelled signaling during Xenopus gastrulation: separable effects on blastopore closure, mesendoderm internalization and archenteron formation

During amphibian gastrulation, the embryo is transformed by the combined actions of several different tissues. Paradoxically, many of these morphogenetic processes can occur autonomously in tissue explants, yet the tissues in intact embryos must interact and be coordinated with one another in order to accomplish the major goals of gastrulation: closure of the blastopore to bring the endoderm and mesoderm fully inside the ectoderm, and generation of the archenteron. Here, we present high-resolution 3D digital datasets of frog gastrulae, and morphometrics that allow simultaneous assessment of the progress of convergent extension, blastopore closure and archenteron formation in a single embryo. To examine how the diverse morphogenetic engines work together to accomplish gastrulation, we combined these tools with time-lapse analysis of gastrulation, and examined both wild-type embryos and embryos in which gastrulation was disrupted by the manipulation of Dishevelled (Xdsh) signaling. Remarkably, although inhibition of Xdsh signaling disrupted both convergent extension and blastopore closure, mesendoderm internalization proceeded very effectively in these embryos. In addition, much of archenteron elongation was found to be independent of Xdsh signaling, especially during the second half of gastrulation. Finally, even in normal embryos, we found a surprising degree of dissociability between the various morphogenetic processes that occur during gastrulation. Together, these data highlight the central role of PCP signaling in governing distinct events of Xenopus gastrulation, and suggest that the loose relationship between morphogenetic processes may have facilitated the evolution of the wide variety of gastrulation mechanisms seen in different amphibian species

Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development

Mitochondrial morphology is determined by a dynamic equilibrium between organelle fusion and fission, but the significance of these processes in vertebrates is unknown. The mitofusins, Mfn1 and Mfn2, have been shown to affect mitochondrial morphology when overexpressed. We find that mice deficient in either Mfn1 or Mfn2 die in midgestation. However, whereas Mfn2 mutant embryos have a specific and severe disruption of the placental trophoblast giant cell layer, Mfn1-deficient giant cells are normal. Embryonic fibroblasts lacking Mfn1 or Mfn2 display distinct types of fragmented mitochondria, a phenotype we determine to be due to a severe reduction in mitochondrial fusion. Moreover, we find that Mfn1 and Mfn2 form homotypic and heterotypic complexes and show, by rescue of mutant cells, that the homotypic complexes are functional for fusion. We conclude that Mfn1 and Mfn2 have both redundant and distinct functions and act in three separate molecular complexes to promote mitochondrial fusion. Strikingly, a subset of mitochondria in mutant cells lose membrane potential. Therefore, mitochondrial fusion is essential for embryonic development, and by enabling cooperation between mitochondria, has protective effects on the mitochondrial population

Cell-cell communication enhances the capacity of cell ensembles to sense shallow gradients during morphogenesis

Collective cell responses to exogenous cues depend on cell-cell interactions. In principle, these can result in enhanced sensitivity to weak and noisy stimuli. However, this has not yet been shown experimentally, and, little is known about how multicellular signal processing modulates single cell sensitivity to extracellular signaling inputs, including those guiding complex changes in the tissue form and function. Here we explored if cell-cell communication can enhance the ability of cell ensembles to sense and respond to weak gradients of chemotactic cues. Using a combination of experiments with mammary epithelial cells and mathematical modeling, we find that multicellular sensing enables detection of and response to shallow Epidermal Growth Factor (EGF) gradients that are undetectable by single cells. However, the advantage of this type of gradient sensing is limited by the noisiness of the signaling relay, necessary to integrate spatially distributed ligand concentration information. We calculate the fundamental sensory limits imposed by this communication noise and combine them with the experimental data to estimate the effective size of multicellular sensory groups involved in gradient sensing. Functional experiments strongly implicated intercellular communication through gap junctions and calcium release from intracellular stores as mediators of collective gradient sensing. The resulting integrative analysis provides a framework for understanding the advantages and limitations of sensory information processing by relays of chemically coupled cells.Comment: paper + supporting information, total 35 pages, 15 figure

GBM heterogeneity as a function of variable epidermal growth factor receptor variant III activity.

Abnormal activation of the epidermal growth factor receptor (EGFR) due to a deletion of exons 2-7 of EGFR (EGFRvIII) is a common alteration in glioblastoma (GBM). While this alteration can drive gliomagenesis, tumors harboring EGFRvIII are heterogeneous. To investigate the role for EGFRvIII activation in tumor phenotype we used a neural progenitor cell-based murine model of GBM driven by EGFR signaling and generated tumor progenitor cells with high and low EGFRvIII activation, pEGFRHi and pEGFRLo. In vivo, ex vivo, and in vitro studies suggested a direct association between EGFRvIII activity and increased tumor cell proliferation, decreased tumor cell adhesion to the extracellular matrix, and altered progenitor cell phenotype. Time-lapse confocal imaging of tumor cells in brain slice cultures demonstrated blood vessel co-option by tumor cells and highlighted differences in invasive pattern. Inhibition of EGFR signaling in pEGFRHi promoted cell differentiation and increased cell-matrix adhesion. Conversely, increased EGFRvIII activation in pEGFRLo reduced cell-matrix adhesion. Our study using a murine model for GBM driven by a single genetic driver, suggests differences in EGFR activation contribute to tumor heterogeneity and aggressiveness

Calcium signaling during convergent extension in Xenopus

Background: During Xenopus gastrulation, cell intercalation drives convergent extension of dorsal tissues. This process requires the coordination of motility throughout a large population of cells. The signaling mechanisms that regulate these movements in space and time remain poorly understood. Results: To investigate the potential contribution of calcium signaling to the control of morphogenetic movements, we visualized calcium dynamics during convergent extension using a calcium-sensitive fluorescent dye and a novel confocal microscopy system. We found that dramatic intercellular waves of calcium mobilization occurred in cells undergoing convergent extension in explants of gastrulating Xenopus embryos. These waves arose stochastically with respect to timing and position within the dorsal tissues. Waves propagated quickly and were often accompanied by a wave of contraction within the tissue. Calcium waves were not observed in explants of the ventral marginal zone or prospective epidermis. Pharmacological depletion of intracellular calcium stores abolished the calcium dynamics and also inhibited convergent extension without affecting cell fate. These data indicate that calcium signaling plays a direct role in the coordination of convergent extension cell movements. Conclusions: The data presented here indicate that intercellular calcium signaling plays an important role in vertebrate convergent extension. We suggest that calcium waves may represent a widely used mechanism by which large groups of cells can coordinate complex cell movements

Cassini ISS Observation of Saturn’s North Polar Vortex and Comparison to the South Polar Vortex

We present analyses of Saturn’s north pole using high-resolution images captured in late 2012 by the Cassini spacecraft’s Imaging Science Subsystem (ISS) camera. The images reveal the presence of an intense cyclonic vortex centered at the north pole. In the red and green visible continuum wavelengths, the north polar region exhibits a cyclonically spiraling cloud morphology extending from the pole to 85°N planetocentric latitude, with a 4700 km radius. Images captured in the methane bands, which sense upper tropospheric haze, show an approximately circular hole in the haze extending up to 1.5° latitude away from the pole. The spiraling morphology and the “eye”-like hole at the center are reminiscent of a terrestrial tropical cyclone. In the System III reference frame (rotation period of 10h39m22.4s, Seidelmann et al. 2007; Archinal et al. 2011), the eastward wind speed increases to about 140 m s^(−1) at 89°N planetocentric latitude. The vorticity is (6.5± 1.5)×10^(−4)s^(−1) at the pole, and decreases to (1.3± 1.2)×10^(−4)s^(−1) at 89°N. In addition, we present an analysis of Saturn’s south polar vortex using images captured in January 2007 to compare its cloud morphology to the north pole. The set of images captured in 2007 includes filters that have not been analyzed before. Images captured in the violet filter (400 nm) also reveal a bright polar cloud. The south polar morphology in 2007 was more smooth and lacked the small clouds apparent around the north pole in 2012. Saturn underwent equinox in August 2009. The 2007 observation captured the pre-equinox south pole, and the 2012 observation captured the post-equinox north pole. Thus, the observed differences between the poles are likely due to seasonal effects. If these differences indeed are caused by seasonal effects, continuing observations of the summer north pole by the Cassini mission should show a formation of a polar cloud that appears bright in short-wavelength filters

The global energy balance of Titan

The global energy budget of planets and their moons is a critical factor to influence the climate change on these objects. Here we report the first measurement of the global emitted power of Titan. Long-term (2004–2010) observations conducted by the Composite Infrared Spectrometer (CIRS) onboard Cassini reveal that the total emitted power by Titan is (2.84 ± 0.01) × 10^(14) watts. Together with previous measurements of the global absorbed solar power of Titan, the CIRS measurements indicate that the global energy budget of Titan is in equilibrium within measurement error. The uncertainty in the absorbed solar energy places an upper limit on the energy imbalance of 6.0%

Gene expression profiling to study racial differences after heart transplantation.

BackgroundThe basis for increased mortality after heart transplantation in African Americans and other non-Caucasian racial groups is poorly defined. We hypothesized that increased risk of adverse events is driven by biologic factors. To test this hypothesis in the Invasive Monitoring Attenuation through Gene Expression (IMAGE) study, we determined whether the event rate of the primary outcome of acute rejection, graft dysfunction, death, or retransplantation varied by race as a function of calcineurin inhibitor (CNI) levels and gene expression profile (GEP) scores.MethodsWe determined the event rate of the primary outcome, comparing racial groups, stratified by time after transplant. Logistic regression was used to compute the relative risk across racial groups, and linear modeling was used to measure the dependence of CNI levels and GEP score on race.ResultsIn 580 patients monitored for a median of 19 months, the incidence of the primary end point was 18.3% in African Americans, 22.2% in other non-Caucasians, and 8.5% in Caucasians (p < 0.001). There were small but significant correlations of race and tacrolimus trough levels to the GEP score. Tacrolimus levels were similar among the races. Of patients receiving tacrolimus, other non-Caucasians had higher GEP scores than the other racial groups. African American recipients demonstrated a unique decrease in expression of the FLT3 gene in response to higher tacrolimus levels.ConclusionsAfrican Americans and other non-Caucasian heart transplant recipients were 2.5-times to 3-times more likely than Caucasians to experience outcome events in the Invasive Monitoring Attenuation through Gene Expression study. The increased risk of adverse outcomes may be partly due to the biology of the alloimmune response, which is less effectively inhibited at similar tacrolimus levels in minority racial groups

Measuring the three-dimensional shear from simulation data, with applications to weak gravitational lensing

We have developed a new three-dimensional algorithm, based on the standard P$^3$M method, for computing deflections due to weak gravitational lensing. We compare the results of this method with those of the two-dimensional planar approach, and rigorously outline the conditions under which the two approaches are equivalent. Our new algorithm uses a Fast Fourier Transform convolution method for speed, and has a variable softening feature to provide a realistic interpretation of the large-scale structure in a simulation. The output values of the code are compared with those from the Ewald summation method, which we describe and develop in detail. With an optimal choice of the high frequency filtering in the Fourier convolution, the maximum errors, when using only a single particle, are about 7 per cent, with an rms error less than 2 per cent. For ensembles of particles, used in typical $N$-body simulations, the rms errors are typically 0.3 per cent. We describe how the output from the algorithm can be used to generate distributions of magnification, source ellipticity, shear and convergence for large-scale structure.Comment: 22 pages, latex, 11 figure

Saturn's Atmosphere at 1–10 Kilometer Resolution

We present images of Saturn from the final phases of the Cassini mission, including images with 0.5 km per pixel resolution, as high as any Saturn images ever taken. Notable features are puffy clouds resembling terrestrial cumulus, shadows indicating cloud height, dome and bowl shaped cloud structures indicating upwelling and downwelling in anticyclones and cyclones respectively, and filaments, which are thread‐like clouds that remain coherent over distances of 20,000 km. From the coherence of the filaments, we give upper bounds on the diffusivity and kinetic energy dissipation. A radiative transfer analysis by Sanz‐Requena et al. (2018) indicates that methane‐band imagery is most useful in determining cloud and haze properties in the 60–250 mbar pressure range. Our methane‐band imagery finds haze in this pressure range covering 64°‐74°planetocentric latitude. Filaments lie within the haze, and cumulus clouds lie below it, but pressure levels are uncertain below the 250 mbar level