On the nature and function of organizers.

Organizers, which comprise groups of cells with the ability to instruct adjacent cells into specific states, represent a key principle in developmental biology. The concept was first introduced by Spemann and Mangold, who showed that there is a cellular population in the newt embryo that elicits the development of a secondary axis from adjacent cells. Similar experiments in chicken and rabbit embryos subsequently revealed groups of cells with similar instructive potential. In birds and mammals, organizer activity is often associated with a structure known as the node, which has thus been considered a functional homologue of Spemann's organizer. Here, we take an in-depth look at the structure and function of organizers across species and note that, whereas the amphibian organizer is a contingent collection of elements, each performing a specific function, the elements of organizers in other species are dispersed in time and space. This observation urges us to reconsider the universality and meaning of the organizer concept

The structural and functional determinants of the Axin and Dishevelled DIX domains.

BACKGROUND: The dishevelled and axin genes encode multi-domain proteins that play key roles in WNT signalling. Dishevelled prevents beta-catenin degradation by interfering with the interaction of beta-catenin with the degradation-mediating Axin-APC-GSK3beta complex. This interference leads to an accumulation of cytoplasmic beta-catenin, which enters the nucleus and interacts with transcription factors that induce expression of Wnt-target genes. Axin, as a component of the degradation-mediating complex, is a potent negative regulator of Wnt signalling, whereas Dishevelled is a potent activator. Both Dishevelled and Axin possess a DIX (Dishevelled/Axin) domain, which mediates protein-protein interactions, specifically homodimerization. RESULTS: An evolutionary trace analysis of DIX domains identified conserved residues which, when mapped onto the crystal structure of the Axin DIX domain and a comparative model of the Dishevelled DIX domain, allow their categorization as residues of either structural or functional importance. We identify residues that are structural and functional determinants of the DIX domain fold, as well as those that are specific to homodimerization of Axin and Dishevelled. CONCLUSION: This report provides the first explanation of the mutant phenotypes caused by non-synonymous substitutions in the Dishevelled and Axin DIX domain by correlating their presumed functional significance with molecular structure.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

Patterned Cell Adhesion Associated with Tissue Deformations during Dorsal Closure in Drosophila

Cell shape changes within epithelia require the regulation of adhesive molecules that maintain tissue integrity. How remodelling of cell contacts is achieved while tissue integrity is maintained remains a fundamental question in morphogenesis. Dorsal Closure is a good system to study the dynamics of DE-Cadherin during morphogenesis. It relies on concerted cell shape changes of two epithelial sheets: amnioserosa cell contraction and epidermal cell elongation. To investigate the modulation of DE-Cadherin we performed antibody uptake experiments in live embryos during Dorsal Closure. We found that some antibodies access certain epitopes of the extracellular domain of native DE-Cadherin only in the amnioserosa and epidermal cells attached to the amnioserosa, which has never been observed in fixed DE-Cadherin in Drosophila embryos. These differences correlate with the different cell behaviour of these regions and therefore we suggest that DE-Cadherin exists in different forms that confer different adhesive strengths. We propose this to be a widespread mechanism for the differential modulation of adhesion during morphogenesis

Pluripotent stem cell models of early mammalian development.

Pluripotent stem cells derived from the early mammalian embryo offer a convenient model system for studying cell fate decisions in embryogenesis. The last 10 years have seen a boom in the popularity of two-dimensional micropatterns and three-dimensional stem cell culture systems as a way to recreate the architecture and interactions of particular cell populations during development. These methods enable the controlled exploration of cellular organization and patterning during development, using cell lines instead of embryos. They have established a new class of in vitro model system for pre-implantation and peri-implantation embryogenesis, ranging from models of the blastocyst stage, through gastrulation and toward early organogenesis. This review aims to set these systems in context and to highlight the strengths and suitability of each approach in modelling early mammalian development.This work was supported by the European Research Council (Advanced Grant to A.M.A., number 834580) and the Leverhulme Trust (RPG-2018-356). N.M. holds the Constance Work Junior Research Fellowship at Newnham College, Cambridge

Neuro-mesodermal progenitors (NMPs): a comparative study between pluripotent stem cells and embryo-derived populations.

The mammalian embryo's caudal lateral epiblast (CLE) harbours bipotent progenitors, called neural mesodermal progenitors (NMPs), that contribute to the spinal cord and the paraxial mesoderm throughout axial elongation. Here, we performed a single cell analysis of different in vitro NMP populations produced either from embryonic stem cells (ESCs) or epiblast stem cells (EpiSCs) and compared them with E8.25 CLE mouse embryos. In our analysis of this region, our findings challenge the notion that NMPs can be defined by the exclusive co-expression of Sox2 and T at mRNA level. We analyse the in vitro NMP-like populations using a purpose-built support vector machine (SVM) based on the embryo CLE and use it as a classification model to compare the in vivo and in vitro populations. Our results show that NMP differentiation from ESCs leads to heterogeneous progenitor populations with few NMP-like cells, as defined by the SVM algorithm, whereas starting with EpiSCs yields a high proportion of cells with the embryo NMP signature. We find that the population from which the Epi-NMPs are derived in culture contains a node-like population, which suggests that this population probably maintains the expression of T in vitro and thereby a source of NMPs. In conclusion, differentiation of EpiSCs into NMPs reproduces events in vivo and suggests a sequence of events for the emergence of the NMP population

Biomedical and societal impacts of in vitro embryo models of mammalian development.

In recent years, a diverse array of in vitro cell-derived models of mammalian development have been described that hold immense potential for exploring fundamental questions in developmental biology, particularly in the case of the human embryo where ethical and technical limitations restrict research. These models open up new avenues toward biomedical advances in in vitro fertilization, clinical research, and drug screening with potential to impact wider society across many diverse fields. These technologies raise challenging questions with profound ethical, regulatory, and social implications that deserve due consideration. Here, we discuss the potential impacts of embryo-like models, and their biomedical potential and current limitations

FGF/MAPK signaling sets the switching threshold of a bistable circuit controlling cell fate decisions in embryonic stem cells.

Intracellular transcriptional regulators and extracellular signaling pathways together regulate the allocation of cell fates during development, but how their molecular activities are integrated to establish the correct proportions of cells with particular fates is not known. Here we study this question in the context of the decision between the epiblast (Epi) and the primitive endoderm (PrE) fate that occurs in the mammalian preimplantation embryo. Using an embryonic stem cell (ESC) model, we discover two successive functions of FGF/MAPK signaling in this decision. First, the pathway needs to be inhibited to make the PrE-like gene expression program accessible for activation by GATA transcription factors in ESCs. In a second step, MAPK signaling levels determine the threshold concentration of GATA factors required for PrE-like differentiation, and thereby control the proportion of cells differentiating along this lineage. Our findings can be explained by a simple mutual repression circuit modulated by FGF/MAPK signaling. This might be a general network architecture to integrate the activity of signal transduction pathways and transcriptional regulators, and serve to balance proportions of cell fates in several contexts.Work in the Martinez Arias lab was funded by an ERC investigator grant. CS was the recipient of an EMBO long-term fellowship, and CS and PR were supported by a Marie Curie fellowship.This is the final version of the article. It first appeared from the Company of Biologists via http://dx.doi.org/10.1242/dev.12753

Autocrine and paracrine Wingless signalling in the Drosophila midgut by both continuous gradient and asynchronous bursts of wingless expression

Wingless (Wg)/ Wnt signalling is a major regulator of homeostasis in both the mammalian andDrosophilaintestine. InDrosophilathe organisation and function of Wingless signalling in the adult intestine remain poorly understood. Here we characterise the pattern of expression ofwg, the stabilisation of its effector Armadillo in the adultDrosophilamidgut, and correlate them with the response of the cells to Wg signalling activation. We show that in normal homeostasis there is a gradient of Wingless signalling in the intestinal stem cell (ISC) and the undifferentiated progenitor cell (enteroblast, EB) populations along the posterior midgut, with a high point at the midgut-hindgut boundary (pylorus). This gradient results from a combination of two sources of Wingless: a distant source outside the epithelium (the pylorus) and a local one from the ISCs and EBs themselves. Altogether, our studies show that Wingless expression and signalling in the epithelium is not continuous, but operates through bursts that occur randomly in space and time.</ns4:p

Three-dimensional stem cell models of mammalian gastrulation.

Gastrulation is a key milestone in the development of an organism. It is a period of cell proliferation and coordinated cellular rearrangement, that creates an outline of the body plan. Our current understanding of mammalian gastrulation has been improved by embryo culture, but there are still many open questions that are difficult to address because of the intrauterine development of the embryos and the low number of specimens. In the case of humans, there are additional difficulties associated with technical and ethical challenges. Over the last few years, pluripotent stem cell models are being developed that have the potential to become useful tools to understand the mammalian gastrulation. Here we review these models with a special emphasis on gastruloids and provide a survey of the methods to produce them robustly, their uses, relationship to embryos, and their prospects as well as their limitations