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Mannose is crucial for mesoderm specification and symmetry breaking in gastruloids

ABSTRACT: Mannose is crucial for mesoderm specification and symmetry breaking in gastruloids

PROVIDER: PRJNA992567 | ENA |

REPOSITORIES: ENA

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Mannose is crucial for mesoderm specification and symmetry breaking in gastruloids

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.

2024-03-11 | GSE236887 | GEO

Mannose is crucial for mesoderm specification and symmetry breaking in gastruloids

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.

2024-01-12 | MSV000093849 | MassIVE

Symmetry breaking and germ layer emergence through optogenetic Wnt activation in gastruloids

Project description:Symmetry breaking and germ layer emergence through optogenetic Wnt activation in gastruloids

| PRJNA981139 | ENA

Single Cell RNA-seq time course of mouse gastruloids at 0h (mESCs), 24h, 48h and 72h post-aggregation

Project description:Single cell transcriptomic study (using 10x Genomics v3 kits) of gastruloids, aggregates of mESCs, at different developmental timepoints (24h, 48h and 72h post-aggregation). mESCs, corresponding to the 0h timepoint, were also sequenced. The aim of this study was to delineate the cell state transition dynamics underlying anteroposterior symmetry breaking and germ layer formation in gastruloids. Gastruloids used in this study were generated from Bra::GFP reporter mESCs (Fehling et al., 2003).

2023-12-04 | E-MTAB-12749 | biostudies-arrayexpress

Symmetry breaking in gastruloid development

Project description:Symmetry breaking in gastruloid development

| PRJNA759809 | ENA

Symmetry breaking in gastruloid development

Project description:Symmetry breaking in gastruloid development

| PRJNA954812 | ENA

Stabilization of cell-cell adhesions prevents symmetry breaking and locks in pluripotency in 3D gastruloids

Project description:Stabilization of cell-cell adhesions prevents symmetry breaking and locks in pluripotency in 3D gastruloids

| PRJNA821461 | ENA

Stabilization of cell-cell adhesions prevents symmetry breaking and locks in pluripotency in 3D gastruloids [mRNASeq3prime]

Project description:Stabilization of cell-cell adhesions prevents symmetry breaking and locks in pluripotency in 3D gastruloids [mRNASeq3prime]

| PRJNA821473 | ENA

Stabilization of cell-cell adhesions prevents symmetry breaking and locks in pluripotency in 3D gastruloids [mRNASeq]

Project description:Stabilization of cell-cell adhesions prevents symmetry breaking and locks in pluripotency in 3D gastruloids [mRNASeq]

| PRJNA821474 | ENA

Multi-axial self-organisation properties of mouse embryonic stem cells into gastruloids [bulk]

Project description:Gastruloids are small aggregates of murine Embryonic Stem (ES) cells stimulated to undergo gastrulation-like symmetry breaking events in vitro (van den Brink, S. C. et al. 2014, Simunovic, M. & Brivanlou, A. H 2017). We analyzed the transcriptomic profiles of gastruloids at different time-points after aggregation (AA), showing that they are capable of organizing a post-occipital pattern of neural, mesodermal and endodermal derivatives that mimics that of the embryo in terms of gene expression.

2018-06-01 | GSE106225 | GEO

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