NRL^{-/-} gene edited human embryonic stem cells generate rod-deficient retinal organoids enriched in S-cone-like photoreceptors

Organoid cultures represent a unique tool to investigate the developmental complexity of tissues like the human retina. NRL is a transcription factor required for the specification and homeostasis of mammalian rod photoreceptors. In Nrl‐deficient mice, photoreceptor precursor cells do not differentiate into rods, and instead follow a default photoreceptor specification pathway to generate S‐cone‐like cells. To investigate whether this genetic switch mechanism is conserved in humans, we used CRISPR/Cas9 gene editing to engineer an NRL‐deficient embryonic stem cell (ESC) line (NRL^{−/−}), and differentiated it into retinal organoids. Retinal organoids self‐organize and resemble embryonic optic vesicles (OVs) that recapitulate the natural histogenesis of rods and cone photoreceptors. NRL−/− OVs develop comparably to controls, and exhibit a laminated, organized retinal structure with markers of photoreceptor synaptogenesis. Using immunohistochemistry and quantitative polymerase chain reaction (qPCR), we observed that NRL^{−/−} OVs do not express NRL, or other rod photoreceptor markers directly or indirectly regulated by NRL. On the contrary, they show an abnormal number of photoreceptors positive for S‐OPSIN, which define a primordial subtype of cone, and overexpress other cone genes indicating a conserved molecular switch in mammals. This study represents the first evidence in a human in vitro ESC‐derived organoid system that NRL is required to define rod identity, and that in its absence S‐cone‐like cells develop as the default photoreceptor cell type. It shows how gene edited retinal organoids provide a useful system to investigate human photoreceptor specification, relevant for efforts to generate cells for transplantation in retinal degenerative diseases

The present and future system for measuring the Atlantic meridional overturning circulation and heat transport

of the global combined atmosphere-ocean heat flux and so is important for the mean climate of the Atlantic sector of the Northern Hemisphere. This meridional heat flux is accomplished by both the Atlantic Meridional Overturning Circulation (AMOC) and by basin-wide horizontal gyre circulations. In the North Atlantic subtropical latitudes the AMOC dominates the meridional heat flux, while in subpolar latitudes and in the subtropical South Atlantic the gyre circulations are also important. Climate models suggest the AMOC will slow over the coming decades as the earth warms, causing widespread cooling in the Northern hemisphere and additional sea-level rise. Monitoring systems for selected components of the AMOC have been in place in some areas for decades, nevertheless the present observational network provides only a partial view of the AMOC, and does not unambiguously resolve the full variability of the circulation. Additional observations, building on existing measurements, are required to more completely quantify the Atlantic meridional heat transport. A basin-wide monitoring array along 26.5°N has been continuously measuring the strength and vertical structure of the AMOC and meridional heat transport since March 31, 2004. The array has demonstrated its ability to observe the AMOC variability at that latitude and also a variety of surprising variability that will require substantially longer time series to understand fully. Here we propose monitoring the Atlantic meridional heat transport throughout the Atlantic at selected critical latitudes that have already been identified as regions of interest for the study of deep water formation and the strength of the subpolar gyre, transport variability of the Deep Western Boundary Current (DWBC) as well as the upper limb of the AMOC, and inter-ocean and intrabasin exchanges with the ultimate goal of determining regional and global controls for the AMOC in the North and South Atlantic Oceans. These new arrays will continuously measure the full depth, basin-wide or choke-point circulation and heat transport at a number of latitudes, to establish the dynamics and variability at each latitude and then their meridional connectivity. Modeling studies indicate that adaptations of the 26.5°N type of array may provide successful AMOC monitoring at other latitudes. However, further analysis and the development of new technologies will be needed to optimize cost effective systems for providing long term monitoring and data recovery at climate time scales. These arrays will provide benchmark observations of the AMOC that are fundamental for assimilation, initialization, and the verification of coupled hindcast/forecast climate models

The North Atlantic subpolar gyre in four high resolution models

The authors present the first quantitative comparison between new velocity datasets and high-resolution models in the North Atlantic subpolar gyre [1/10° Parallel Ocean Program model (POPNA10), Miami Isopycnic Coordinate Ocean Model (MICOM), ° Atlantic model (ATL6), and Family of Linked Atlantic Ocean Model Experiments (FLAME)]. At the surface, the model velocities agree generally well with World Ocean Circulation Experiment (WOCE) drifter data. Two noticeable exceptions are the weakness of the East Greenland coastal current in models and the presence in the surface layers of a strong southwestward East Reykjanes Ridge Current. At depths, the most prominent feature of the circulation is the boundary current following the continental slope. In this narrow flow, it is found that gridded float datasets cannot be used for a quantitative comparison with models. The models have very different patterns of deep convection, and it is suggested that this could be related to the differences in their barotropic transport at Cape Farewell. Models show a large drift in watermass properties with a salinization of the Labrador Sea Water. The authors believe that the main cause is related to horizontal transports of salt because models with different forcing and vertical mixing share the same salinization problem. A remarkable feature of the model solutions is the large westward transport over Reykjanes Ridge [10 Sv (Sv ≡ 106 m3 s−1) or more

A diagnosis of isopycnal mixing by mesoscale eddies

Combining the buoyancy and tracer budget in the generalised Temporal Residual Mean (TRM-G) framework of [Eden, C., Greatbatch, R.J., Olbers, D. 2007a. Interpreting eddy fluxes. J. Phys. Oceanogr. 37, 1282–1296], we show that within the small slope approximation and weakly diabatic situation, the isopycnal diffusivity is related to the difference of the streamfunctions of the eddy-induced velocities of tracer and buoyancy divided by the angle between the (negative) slopes of isopycnals and the isolines of the tracer. Using this result tracer simulations of a realistic mesoscale-eddy-permitting model of the North Atlantic coupled to a biogeochemical model are diagnosed in terms of zonal (View the MathML sourceKI(x)) and meridional (View the MathML sourceKI(y)) isopycnal diffusivities relevant for non-eddy-permitting ocean models. We find for tracers having different interior sources and surface forcing and therefore different lateral and vertical mean gradients, values of View the MathML sourceKI(x) and View the MathML sourceKI(y) with similar magnitudes and lateral and vertical structure. In general, isopycnal diffusivities lie within the expected range between 0 and 5000 m2/s but we also find a strong anisotropy with View the MathML sourceKI(x) much larger than View the MathML sourceKI(y) over large regions of the North Atlantic. Both View the MathML sourceKI(x) and View the MathML sourceKI(y) are larger within and above the thermocline but decay almost to zero below. Our results also support the common practise of the use of identical isopycnal and thickness diffusivity for any tracer in ocean models

A diagnosis of thickness fluxes in an eddy-resolving model

Output from an eddy-resolving model of the North Atlantic Ocean is used to estimate values for the thickness diffusivity κ appropriate to the Gent and McWilliams parameterization. The effect of different choices of rotational eddy fluxes on the estimated κ is discussed. Using the raw fluxes (no rotational flux removed), large negative values (exceeding −5000 m2 s−1) of κ are diagnosed locally, particularly in the Gulf Stream region and in the equatorial Atlantic. Removing a rotational flux based either on the suggestion of Marshall and Shutts or the more general theory of Medvedev and Greatbatch leads to a reduction of the negative values, but they are still present. The regions where κ < 0 correspond to regions where eddies are acting to increase, rather than decrease (as in baroclinic instability) the mean available potential energy. In the subtropical gyre, κ ranges between 500 and 2000 m2 s−1, rapidly decreasing to zero below the thermocline in all cases. Rotational fluxes and κ are also estimated using an optimization technique. In this case, |κ| can be reduced or increased by construction, but the regions where κ < 0 are still present and the optimized rotational fluxes also remain similar to a priori values given by the theoretical considerations. A previously neglected component (ν) of the bolus velocity is associated with the horizontal flux of buoyancy along, rather than across, the mean buoyancy contours. The ν component of the bolus velocity is interpreted as a streamfunction for eddy-induced advection, rather than diffusion, of mean isopycnal layer thickness, showing up when the lateral eddy fluxes cannot be described by isotropic diffusion only. All estimates show a similar large-scale pattern for ν, implying westward advection of isopycnal thickness over much of the subtropical gyre. Comparing ν with a mean streamfunction shows that it is about 10% of the mean in midlatitudes and even larger than the mean in the Tropics

Wnt/beta-catenin promotes gastric fundus specification in mice and humans

Despite the global prevalence of gastric disease, there are few adequate models in which to study the fundus epithelium of the human stomach. We differentiated human pluripotent stem cells (hPSCs) into gastric organoids containing fundic epithelium by first identifying and then recapitulating key events in embryonic fundus development. We found that disruption of Wnt/β-catenin signalling in mouse embryos led to conversion of fundic to antral epithelium, and that β-catenin activation in hPSC-derived foregut progenitors promoted the development of human fundic-type gastric organoids (hFGOs). We then used hFGOs to identify temporally distinct roles for multiple signalling pathways in epithelial morphogenesis and differentiation of fundic cell types, including chief cells and functional parietal cells. hFGOs are a powerful model for studying the development of the human fundus and the molecular bases of human gastric physiology and pathophysiology, and also represent a new platform for drug discovery

Eddy-Permitting Ocean Circulation Hindcasts of Past Decades

International audienc

A model of multiple zonal jets in the oceans : dynamical and kinematical analysis

Author Posting. © American Meteorological Society, 2009. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 39 (2009): 2711-2734, doi:10.1175/2009JPO4093.1.Multiple alternating zonal jets observed in the ocean are studied with an idealized quasigeostrophic zonal-channel model, with the supercritical, zonal background flow imposed. Both eastward and westward background flows with vertical shear are considered. The underlying nonlinear dynamics is illuminated with analysis of the vertical-mode interactions and time-mean eddy fluxes. Interactions between the vertical modes are systematically studied. The barotropic component of the jets is maintained by both barotropic–barotropic and baroclinic–baroclinic time-mean interactions; thus, the barotropic component of the jets cannot be accurately simulated with a randomly forced barotropic model. The roles of the vertical-mode interactions in driving the baroclinic component of the jets are also characterized. Not only the first but also the second baroclinic mode is found to be important for maintaining the baroclinic component of the jets, whereas the barotropic component of the jets is maintained mostly by the barotropic and first baroclinic modes. The properties of the eddy forcing were systematically studied. It is shown that the baroclinic component of the jets is maintained by Reynolds stress forcing and resisted by form stress forcing only in the eastward background flow. In the westward background flow, the jets are maintained by form stress forcing and resisted by Reynolds stress forcing. The meridional scaling and kinematical properties of the jets are studied as well as the roles of meridional boundaries. The Rhines scaling for meridional spacing of the jets is not generally confirmed, and it is also shown that there are multiple stable equilibria with different numbers of the time-mean jets. It is also found that the jets are associated with alternating weak barriers to the meridional material transport, but the locations of these barriers are not unique and depend on the direction of the background flow and depth. Finally, if the channel is closed with meridional walls, then the jets become more latent but the eddy forcing properties do not change qualitatively.Funding for PB was provided by NSF Grants OCE 0344094 and OCE 0725796 and by the research grant from the Newton Trust of the University of Cambridge. Funding for IK was provided by NSF Grants OCE 0346178 and 0749722. Funding for JP was provided by NSF Grant OCE 0451086