Mesoderm specification under hypoxia
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA721233 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14209432_1.fastq.gz | Fastqsanger.gz | |||
| SRR14209432_2.fastq.gz | Fastqsanger.gz | |||
| SRR14209433_1.fastq.gz | Fastqsanger.gz | |||
| SRR14209433_2.fastq.gz | Fastqsanger.gz | |||
| SRR14209434_1.fastq.gz | Fastqsanger.gz |
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Project description:AB2.2 cells were subjected to hanging-drop differentiation under normoxia and hypoxia, respectively. RNA-seq were performed at differentiation day 4, when mesoderm markers peaked. Three biological replicates were set for both groups.The expression matrix was obtained by Hisat2 followed by Stringtie.
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Project description:Patterning and growth are fundamental features of embryonic development that must be tightly coordinated during morphogenesis. While metabolism is known to control cell growth, how it impacts patterning and links to morphogenesis is poorly understood. To understand how metabolism impacts early mesoderm specification during gastrulation, we used in vitro mouse embryonic stem (ES) cell-derived gastruloids, due to ease of metabolic manipulations and high-throughput nature. Gastruloids showed mosaic expression of glucose transporters co-expressing with the mesodermal marker T/Bra. To understand the significance of cellular glucose uptake in development, we used the glucose metabolism inhibitor 2-deoxy-D-glucose (2-DG). 2-DG blocked the expression of T/Bra and abolishes axial elongation in gastruloids. Surprisingly, removing glucose completely from the medium did not phenocopy 2-DG treatment despite a significant decline in glycolytic intermediates occurring under both conditions. As 2-DG can also act as a competitive inhibitor of mannose in protein glycosylation, we added mannose together with 2-DG and found that it could rescue the mesoderm specification. We corroborated these results in vivomouse embryos where supplementing mannose rescued the 2-DG mediated phenotype of mesoderm specification and proximo-distal elongation of the primitive streak. We further showed that blocking production and intracellular recycling of mannose abrogated mesoderm specification. At molecular level, proteomics analysis revealed that mannose reversed glycosylation of the Wnt pathway regulator, Secreted Frizzled Receptor, Frzb, expressed in the primitive streak of the mouse embryo. Our study showed how mannose linked metabolism to glycosylation of a developmental pathway component, crucial in patterning of mesoderm and morphogenesis of gastruloids.
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Project description:Mbodj2016 - Mesoderm specification during
Drosophila development
This model is described in the article:
Qualitative Dynamical
Modelling Can Formally Explain Mesoderm Specification and
Predict Novel Developmental Phenotypes.
Mbodj A, Gustafson EH, Ciglar L,
Junion G, Gonzalez A, Girardot C, Perrin L, Furlong EE, Thieffry
D.
PLoS Comput. Biol. 2016 Sep; 12(9):
e1005073
Abstract:
Given the complexity of developmental networks, it is often
difficult to predict the effect of genetic perturbations, even
within coding genes. Regulatory factors generally have
pleiotropic effects, exhibit partially redundant roles, and
regulate highly interconnected pathways with ample cross-talk.
Here, we delineate a logical model encompassing 48 components
and 82 regulatory interactions involved in mesoderm
specification during Drosophila development, thereby providing
a formal integration of all available genetic information from
the literature. The four main tissues derived from mesoderm
correspond to alternative stable states. We demonstrate that
the model can predict known mutant phenotypes and use it to
systematically predict the effects of over 300 new, often
non-intuitive, loss- and gain-of-function mutations, and
combinations thereof. We further validated several novel
predictions experimentally, thereby demonstrating the
robustness of model. Logical modelling can thus contribute to
formally explain and predict regulatory outcomes underlying
cell fate decisions.
This model is hosted on
BioModels Database
and identified by:
MODEL1607310000.
To cite BioModels Database, please use:
BioModels Database:
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To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
2016-10-25 | MODEL1607310000 | BioModels
Project description:Specification of embryonic mesoderm by Pinhead signaling
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Project description:Patterning and growth are fundamental features of embryonic development that must be tightly coordinated during morphogenesis. As metabolism can control cell growth while also providing mechanistic links to developmental signalling pathways, it is ideally placed to enable this coordination. To understand how metabolism impacts early mesoderm specification, we used mouse embryonic stem (ES) cell-derived gastruloids, as these enable temporal control over metabolic manipulations and can be generated in large quantities. Gastruloids show mosaic expression of two glucose transporters,Slc2a1andSlc2a3both of which co-express with the expression of both the mesodermal markerT/Braand the neural markerSox2. To understand the significance of cellular glucose uptake in development, we used the glucose metabolism inhibitor 2-deoxy-D-glucose (2-DG). 2-DG specifically blocks the expression ofT/Brawithout affecting the expression ofSox2and abolishes axial elongation in gastruloids. Surprisingly, removing glucose completely from the medium did not phenocopy 2-DG treatment despite a significant decline in glycolytic intermediates occurring under both conditions. As 2-DG can also act as a competitive inhibitor of mannose, we added mannose together with 2-DG and found that it could rescue the mesoderm specification. Together, our results show that while mannose is crucial for mesoderm specification, the glycolytic pathway is dispensable at early stages ofT/Braexpression in gastruloids.
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Project description:Cardiopharyngeal mesoderm contributes to the formation of the heart and head muscles. However, the mechanisms governing cardiopharyngeal mesoderm specification remain unclear. Indeed, there is a lack of an in vitro model replicating the differentiation of both heart and head muscles to study these mechanisms. Such models are required to allow live-imaging and high throughput genetic and drug screening. Here, we show that the formation of self-organizing or pseudo-embryos from mouse embryonic stem cells (mESCs), also called gastruloids, reproduces cardiopharyngeal mesoderm specification towards cardiac and skeletal muscle lineages. By conducting a comprehensive temporal analysis of cardiopharyngeal mesoderm establishment and differentiation in gastruloids and comparing it to mouse embryos, we present the first evidence for skeletal myogenesis in gastruloids. By inferring lineage trajectories from the gastruloids single-cell transcriptomic data, we further suggest that heart and head muscles formed in gastruloids derive from cardiopharyngeal mesoderm progenitors. We identify different subpopulations of cardiomyocytes and skeletal muscles, which most likely correspond to different states of myogenesis with “head-like” and “trunk-like” skeletal myoblasts. These findings unveil the potential of mESC-derived gastruloids to undergo specification into both cardiac and skeletal muscle lineages, allowing the investigation of the mechanisms of cardiopharyngeal mesoderm differentiation in development and how this could be affected in congenital diseases.
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Project description:Among the three embryonic germ layers, mesoderm plays a central role in the establishment of the vertebrate body plan. Mesoderm is specified by secreted signaling proteins from the FGF, Nodal, BMP and Wnt families. No new classes of extracellular mesoderm-inducing factors have been identified in more than two decades. Here we show that the Xenopus pinhead (pnhd) gene encodes a novel secreted protein that activates specific mesodermal markers in presumptive ectoderm. We demonstrate that Pnhd is required for the specification of a subset of embryonic mesoderm in vivo. RNA sequencing revealed that many transcriptional targets of Pnhd are shared with those of the FGF pathway. Pnhd signaling involves active FGF but not Wnt receptors and is accompanied by Erk1 phosphorylation. We propose that Pnhd operates in the marginal zone to specify mesodermal progenitors via an FGF receptor-dependent mechanism.
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