Englacial Pore Water Localizes Shear in Temperate Ice Stream Margins

The margins of fast‐moving ice streams are characterized by steep velocity gradients. Some of these gradients cannot be explained by a temperature‐dependent viscosity alone. Laboratory data suggest that water in the ice‐grain matrix decreases the ice viscosity; we propose that this causes the strong localization of shear in temperate ice stream margins. However, the magnitude of weakening and its consequences for ice stream dynamics are poorly understood. Here we investigate how the coupling between temperate ice properties, ice mechanics, and drainage of melt water from the ice stream margin alters the dynamics of ice streams. We consider the steady‐state ice flow, temperature, water content, and subglacial water drainage in an ice stream cross section. Temperate ice dynamics are modeled as a two‐phase flow, with gravity‐driven water transport in the pores of a viscously compacting and deforming ice matrix. We find that the dependence of ice viscosity on meltwater content focuses the temperate ice region and steepens the velocity gradients in the ice stream margin. It provides a possible explanation for the steep velocity gradients observed in some ice stream shear margins. This localizes heat dissipation there, which in turn increases the amount of meltwater delivered to the ice stream bed. This process is controlled by the permeability of the temperate ice and the sensitivity of ice viscosity to meltwater content, both of which are poorly constrained properties

Oscillatory subglacial drainage in the absence of surface melt

The presence of strong diurnal cycling in basal water pressure records obtained during the melt season is well established for many glaciers. The behaviour of the drainage system outside the melt season is less well understood. Here we present borehole observations from a surge-type valley glacier in the St Elias Mountains, Yukon Territory, Canada. Our data indicate the onset of strongly correlated multi-day oscillations in water pressure in multiple boreholes straddling a main drainage axis, starting several weeks after the disappearance of a dominant diurnal mode in August 2011 and persisting until at least January 2012, when multiple data loggers suffered power failure. Jökulhlaups provide a template for understanding spontaneous water pressure oscillations not driven by external supply variability. Using a subglacial drainage model, we show that water pressure oscillations can also be driven on a much smaller scale by the interaction between conduit growth and distributed water storage in smaller water pockets, basal crevasses and moulins, and that oscillations can be triggered when water supply drops below a critical value. We suggest this in combination with a steady background supply of water from ground water or englacial drainage as a possible explanation for the observed wintertime pressure oscillations

An indelible lineage marker for Xenopus using a mutated green fluorescent protein

We describe the use of a DNA construct (named GFP.RN3) encoding green fluorescent protein as a lineage marker for Xenopus embryos. This offers the following advantages over other lineage markers so far used in Xenopus. When injected as synthetic mRNA, its protein emits intense fluorescence in living embryos. It is non-toxic, and the fluorescence does not bleach when viewed under 480 nm light. It is surprisingly stable, being strongly visible up to the feeding tadpole stage (5 days), and in some tissues for several weeks after mRNA injection. We also describe a construct that encodes a blue fluorescent protein. We exemplify the use of this GFP.RN3 construct for marking the lineage of individual blastomeres at the 32- to 64-cell stage, and as a marker for single transplanted blastula cells. Both procedures have revealed that the descendants of one embryonic cell can contribute single muscle cells to nearly all segmental myotomes rather than predominantly to any one myotome. An independent aim of our work has been to follow the fate of cells in which an early regulatory gene has been temporarily overexpressed. For this purpose, we co-injected GFP.RN3 mRNA and mRNA for the early Xenopus gene Eomes, and found that a high concentration of Eomes results in ectopic muscle gene activation in only the injected cells. This marker may therefore be of general value in providing long term identification of those cells in which an early gene with ephemeral expression has been overexpressed

Following cell fate in the living mouse embryo

It has been difficult to follow many of the dramatic changes in cell fate and cell migration during mouse development. This is because there has been no enduring marker that would allow cells to be recognised in the living embryo. We believe that we have overcome this problem by developing a novel form of green fluorescent protein, named MmGFP, that proves to be easily visible and non toxic to mouse cells and does not perturb embryogenesis. We show that synthetic mRNA encoding MmGFP can be injected into blastomeres to follow the fate of their progeny during preimplantation development. We have made a stable embryonic stem cell line that expresses MmGFP and introduced these fluorescent cells into mouse embryos. For the first time, we have been able to follow the fate of embryonic stem cells in living embryos and to observe directly the contribution of these cells to distinct lineages of the postimplantation embryo. This approach should lead to a more complete description of the dynamics of cell fate in the mouse

GAL4 GFP enhancer trap lines for analysis of stomatal guard cell development and gene expression.

To facilitate the monitoring of guard cells during development and isolation, a population of 704 GAL4 GFP enhancer trap lines was screened and four single insert lines with guard cell GFP expression and one with developmentally-regulated guard cell GFP expression were identified. The location of the T-DNA inserts, the expression of the flanking genes, and the promoter activity of the genomic DNA upstream of the T-DNA were characterized. The results indicated that the GFP expression pattern in at least one of the lines was due to elements in the intergenic DNA immediately upstream of the T-DNA, rather than due to the activity of the promoters of genes flanking the insert, and provide evidence for the involvement of Dof elements in regulating guard cell gene expression. It is shown further that the GAL4 GFP lines can be used to track the contribution of guard cell material in vitro, and this method was used to assess the purity of guard cell samples obtained using two methods of guard cell isolation

Characterization of Intrinsic Properties of Promoters.

Accurate characterization of promoter behavior is essential for the rational design of functional synthetic transcription networks such as logic gates and oscillators. However, transcription rates observed from promoters can vary significantly depending on the growth rate of host cells and the experimental and genetic contexts of the measurement. Furthermore, in vivo measurement methods must accommodate variation in translation, protein folding, and maturation rates of reporter proteins, as well as metabolic load. The external factors affecting transcription activity may be considered to be extrinsic, and the goal of characterization should be to obtain quantitative measures of the intrinsic characteristics of promoters. We have developed a promoter characterization method that is based on a mathematical model for cell growth and reporter gene expression and exploits multiple in vivo measurements to compensate for variation due to extrinsic factors. First, we used optical density and fluorescent reporter gene measurements to account for the effect of differing cell growth rates. Second, we compared the output of reporter genes to that of a control promoter using concurrent dual-channel fluorescence measurements. This allowed us to derive a quantitative promoter characteristic (ρ) that provides a robust measure of the intrinsic properties of a promoter, relative to the control. We imposed different extrinsic factors on growing cells, altering carbon source and adding bacteriostatic agents, and demonstrated that the use of ρ values reduced the fraction of variance due to extrinsic factors from 78% to less than 4%. This is a simple and reliable method to quantitatively describe promoter properties.TJR was supported by a Microsoft Research studentship and EC FP7 Project No. 612146 (PLASWIRES) awarded to JH, JRB by a Microsoft Research studentship and internship, and FF by CONICYT-PAI/Concurso Nacional de Apoyo al Retorno de Investigadores/as desde el Extranjero Folio 8213002 7, and EPSRC grant EP/H019162/1 awarded to JH. JWA acknowledges the EPSRC and the Wellcome Trust for support.This is the author accepted manuscript. The final version is available from ACS via http://dx.doi.org/10.1021/acssynbio.5b0011

Droplet-based microfluidic analysis and screening of single plant cells.

Droplet-based microfluidics has been used to facilitate high-throughput analysis of individual prokaryote and mammalian cells. However, there is a scarcity of similar workflows applicable to rapid phenotyping of plant systems where phenotyping analyses typically are time-consuming and low-throughput. We report on-chip encapsulation and analysis of protoplasts isolated from the emergent plant model Marchantia polymorpha at processing rates of >100,000 cells per hour. We use our microfluidic system to quantify the stochastic properties of a heat-inducible promoter across a population of transgenic protoplasts to demonstrate its potential for assessing gene expression activity in response to environmental conditions. We further demonstrate on-chip sorting of droplets containing YFP-expressing protoplasts from wild type cells using dielectrophoresis force. This work opens the door to droplet-based microfluidic analysis of plant cells for applications ranging from high-throughput characterisation of DNA parts to single-cell genomics to selection of rare plant phenotypes.BBSRC and EPSRC OpenPlant BB/L014130/

A low-power data acquisition system for geomagnetic observatories and variometer stations

A modern geomagnetic observatory must provide data of high stability, continuity, and resolution. The INTERMAGNET network has therefore specified quantitative criteria to ensure a high quality standard of geomagnetic observatories. Here, we present a new data acquisition system which was designed to meet these criteria, in particular with respect to 1 Hz data. This system is based on a Raspberry Pi embedded PC and runs a C+ +  data acquisition software. As a result, the data acquisition system is modular, cheap, and flexible, and it can be operated in remote areas with limited power supply. In addition, the system is capable of near-real-time data transmission, using a reverse SSH tunnel to work with any network available. The system hardware was successfully tested at the Niemegk observatory for a period of 1 year and subsequently installed at the Tatuoca observatory in Brazil

New tools for self-organized pattern formation

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