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Dual role of Ovol2 on the germ cell lineage segregation during gastrulation in mouse embryogenesis.

ABSTRACT: Dual role of Ovol2 on the germ cell lineage segregation during gastrulation in mouse embryogenesis.

PROVIDER: PRJNA765589 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16019700.fastq.gz	Fastqsanger.gz
	SRR16019701.fastq.gz	Fastqsanger.gz
	SRR16019702.fastq.gz	Fastqsanger.gz
	SRR16019703.fastq.gz	Fastqsanger.gz
	SRR16019704.fastq.gz	Fastqsanger.gz

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Dual role of Ovol2 on the germ cell lineage segregation during gastrulation in mouse embryogenesis. [RNA-Seq]

Project description:In mammals, primordial germ cells (PGCs), the origin of the germ line, are specified from the epiblast at the posterior region where gastrulation simultaneously occurs, yet the functional relationship between PGC specification and gastrulation remains unclear. Here, we show that Ovol2, a transcription factor conserved across the animal kingdom, balances these major developmental processes by repressing the epithelial-to-mesenchymal transition (EMT) driving gastrulation and the upregulation of genes associated with PGC specification. Ovol2a, a splice variant encoding a repressor domain, directly regulates EMT-related genes and consequently induces re-acquisition of potential pluripotency during PGC specification, whereas Ovol2b, another splice variant missing the repressor domain, directly upregulates genes associated with PGC specification. Taken together, these results elucidate the molecular mechanism underlying allocation of the germ line among epiblast cells differentiating into somatic cells through gastrulation.

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Dual role of Ovol2 on the germ cell lineage segregation during gastrulation in mouse embryogenesis. [ChIP-Seq]

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Dual role of Ovol2 on the germ cell lineage segregation during gastrulation in mouse embryogenesis. [ChIP-Seq]

Project description:Dual role of Ovol2 on the germ cell lineage segregation during gastrulation in mouse embryogenesis. [ChIP-Seq]

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Dual role of Ovol2 on the germ cell lineage segregation during gastrulation in mouse embryogenesis. [RNA-Seq]

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Dual role of Ovol2 on the germ cell lineage segregation during gastrulation in mouse embryogenesis.

Project description:This SuperSeries is composed of the SubSeries listed below.

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Ex utero Mouse Embryogenesis from Pre-Gastrulation to Late-Organogenesis

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Project description:Establishment of the mammalian body plan occurs shortly after the embryo implants into the maternal uterus, and our understanding of post-implantation developmental processes remains limited. While methods for in vitro culture of pre- and peri-implantation mouse embryos are routinely utilized, approaches for robust culture of post-implantation embryos from egg cylinder stages until advanced organogenesis remain to be established. We develop herein highly stable ex utero post-implantation mouse embryo culture platforms, that enable appropriate development of embryos before gastrulation (E5.5) until the hind limb formation stage (E11). Late gastrulating embryos (E7.5) are grown in 3D rotating bottles settings, while extended culture from pre-gastrulation stages (E5.5 or E6.5) requires a combination of novel static and rotating bottle culture protocols. Histological, molecular, and single cell RNA-seq analysis validate that the ex utero developed embryos recapitulate precisely in utero development. This culture system is amenable to introducing a variety of embryonic perturbations and micro-manipulations that can be followed ex utero for up to 6 days. Establishment of a system to robustly grow normal mouse embryos ex utero from pre-gastrulation to advanced organogenesis represents a valuable tool to investigate post-implantation embryogenesis, eliminating the uterine barrier to mechanistically interrogate morphogenesis and tissue specification in mammals.

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Project description:Gastrulation is a critical stage of embryonic development during which the three germ layers are established. Deciphering the molecular mechanisms underlying this process from a protein perspective remains a significant challenge. To address this, we employed a multilayered mass spectrometry-based proteomics approach to investigate the global dynamics of (phospho)protein expression during differentiation of ESCs towards gastruloids – an in vitro model of gastrulation-stage embryogenesis. Our findings revealed that many proteins exhibited temporal expression during gastrulation with unique expression profiles corresponding to the three germ layers, which we also validated using an ultra sensitive single cell proteomics approachtechnology. Additionally, we profiled enhancer interaction landscapes in ESCs and gastruloids using p300 proximity labeling, which revealed numerous gastruloid-specific transcription factors and chromatin remodelers. Subsequent degron based perturbations combined with scRNA-seq revealed a critical role for Zeb2 in regulating mouse and human somitogenesis. Overall, this study provides a rich resource for developmental and synthetic biology communities endeavoring to understand mammalian embryogenesis.

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Project description:Anterior mesoderm (AM) and definitive endoderm (DE) progenitors represent the earliest embryonic cell types that are specified during germ layer formation at the primitive streak (PS) of the mouse embryo. Genetic experiments indicate that both lineages segregate from Eomes expressing progenitors in response to different NODAL signaling levels. However, the precise spatiotemporal pattern of the emergence of these cell types and molecular details of lineage segregation remain unexplored. We combined genetic fate labeling and imaging approaches with single cell RNA sequencing (scRNA-seq) to follow the transcriptional identities and define lineage trajectories of Eomes dependent cell types. Accordingly, all cells moving through the PS during the first day of gastrulation express Eomes AM and DE specification occurs before cells leave the PS from Eomes positive progenitors in a distinct spatiotemporal pattern. ScRNA-seq analysis further suggest the immediate and complete separation of AM and DE lineages from Eomes expressing cells as last common bipotential progenitor.

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