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Embryonic stem cells derived from developmentally incompetent embryos deficient in maternal and zygotic Cdx2

ABSTRACT: Separation of cell lineages during early mammalian development is required to establish the pluripotent founder cell population that will give rise to the embryo proper and a functional trophoblast to support its development. We systemically assessed the role of the homeobox gene Cdx2 in vivo and in vitro development with an RNAi approach. Effective elimination of both maternal and zygotic Cdx2 resulted in typical phenotypes of Cdx2-mutant embryos, such as failure of hatching and implantation. However, the blastulation and expression of TE specific markers in these Cdx2-deficient embryos excluded the possibility of Cdx2 to act as a TE determinant, although compromised structure and functioning of TE was observed and the resulted embryos were not viable. Strikingly, the efficiency of stem cell derivation was significantly higher than control when embryos were put on MEF at the 8-cell stage and the derived stem cells were fully pluripotent as shown by chimera and tetraploid complementation experiments. Comparative genomic hybridization of wild type and Cdx2 mutant at 8-cell and blastocyst mouse embryos were performed. 8-cell biological duplicates and blastocyst stage biological triplicates embryos were used.The hybridization experiments were duplicated in a reciprocal labeling manner to reduce dye integration bias (dye-swaps).

ORGANISM(S): Mus musculus

SUBMITTER: Marcos Arauzo-Bravo

PROVIDER: E-GEOD-12330 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Embryonic stem cells derived from developmentally incompetent embryos deficient in maternal and zygotic Cdx2

Project description:Separation of cell lineages during early mammalian development is required to establish the pluripotent founder cell population that will give rise to the embryo proper and a functional trophoblast to support its development. We systemically assessed the role of the homeobox gene Cdx2 in vivo and in vitro development with an RNAi approach. Effective elimination of both maternal and zygotic Cdx2 resulted in typical phenotypes of Cdx2-mutant embryos, such as failure of hatching and implantation. However, the blastulation and expression of TE specific markers in these Cdx2-deficient embryos excluded the possibility of Cdx2 to act as a TE determinant, although compromised structure and functioning of TE was observed and the resulted embryos were not viable. Strikingly, the efficiency of stem cell derivation was significantly higher than control when embryos were put on MEF at the 8-cell stage and the derived stem cells were fully pluripotent as shown by chimera and tetraploid complementation experiments. Comparative genomic hybridization of wild type and Cdx2 mutant at 8-cell and blastocyst mouse embryos were performed.

2008-12-30 | GSE12330 | GEO

Functional role of GATA3 and CDX2 in lineage specification during bovine early embryonic development

Project description:Current understandings of the initiation of the trophectoderm (TE) program in mammalian embryonic development lacks evidence of how TE-associated factors such as CDX2 and GATA3 participate in bovine lineage specification. In this study, we describe the effects of TE-associated factors on lineage specification marker genes such as SOX2, OCT4, NANOG, GATA6 and SOX17, assisted by a cytosine base editor system. Interestingly, GATA3 downregulates the NANOG expression in bovine blastocysts. Further analysis of the blastocyst mosaic shows that GATA3 is required for NANOG in TE cells of bovine blastocysts. It is worthy to notice that, unlike mouse embryos where GATA3 and CDX2 did not reciprocally affect each other in bovine embryos.

2022-10-26 | GSE216123 | GEO

TEAD4 role in trophectoderm commitment and development is not conserved in non-rodent mammals

Project description:The first lineage differentiation in mammals gives rise to the inner cell mass (ICM) and the trophectoderm (TE). In mice, TEAD4 is a master regulator of TE commitment, as it regulates the expression of other TE-specific genes and its ablation prevents blastocyst formation, but its role in other mammals remains unclear. TEAD4 ablation in bovine embryos did not impede TE differentiation or blastocyst formation, but a transcriptomic analysis was performed to see if there were any transcriptomic anomalies in knockout embryos.

2024-05-05 | GSE266304 | GEO

Genome-wide mapping of Eset-binding sites and H3K9me3 state in mouse embryonic stem cells

Project description:The histone H3 lysine 9 (H3K9) methyltransferase Eset is an epigenetic regulator critical for the development of the inner cell mass (ICM). Although ICM-derived embryonic stem (ES) cells are normally unable to contribute to the trophectoderm (TE) in blastocysts, we find that depletion of Eset by shRNAs leads to differentiation with the formation of trophoblast-like cells and induction of trophoblast-associated gene expression. Using ChIP-seq analyses, we identified Eset target genes with Eset-dependent H3K9 trimethylation. We confirmed that genes that are preferentially expressed in the TE (Tcfap2a and Cdx2) are bound and repressed by Eset. Single cell PCR analysis shows that the expression of Cdx2 and Tcfap2a is also induced in Eset-depleted morula cells. Importantly, Eset-depleted cells can incorporate into the TE of a blastocyst and subsequently placental tissues. Co-immunoprecipitation and ChIP assays further demonstrates that Eset interacts with Oct4, which in turn recruits Eset to silence these trophoblast-associated genes. Our result suggests that Eset restricts the extraembryonic trophoblast lineage potential of pluripotent cells and links an epigenetic regulator to key cell fate decision through a pluripotency factor. Keywords: Epigenetics Examine Eset-binding sites and compare the H3K9me3 state between Eset knockdown mouse ES cells and control knockdown mouse ES cells.

2009-11-04 | E-GEOD-17642 | biostudies-arrayexpress

Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency

Project description:Proper lineage specification is extremely important for the embryo's implantation and development. Numerous findings of lineage specification emerged in the mouse embryo but remain unclear in the bovine embryo due to the limited techniques. Here, our study is the first to demonstrate that the single base editor (ABE7.10 and BE3) can be applied to bovine embryos for base conversion, which contributes us to further exploring the lineage specification of the bovine embryo. We further show SOX2 is not necessary for blastocyst formation but is required for ICM maintenance in bovine early embryo development. The underlying mechanism may be that SOX2 promoted the differentiation of ICM by maintaining the expression levels of OCT4 and NANOG. Meanwhile, the disappearance of SOX2 in TE of bovine late blastocyst depended on the expression of CDX2.

2022-03-22 | GSE189318 | GEO

RNA-sequencing technology identifies specific genes expressed in the ICM of mouse embryos

Project description:RNA-seq data from whole mouse embryos at E3.75 (stage where the three cell types: TE, PrE and EPI are well resolved) and from dissected ICMs in order to identify genes expressed specifically in the ICM We used E3.75 whole mouse embryos at the blastocyst stage and immunosurgically dissected ICMs for RNA extraction and sequencing

2013-04-23 | E-GEOD-44553 | biostudies-arrayexpress

Preimplantation mouse embryo proteome during 8C to 16C division

Project description:Preimplantation stages of mouse embryo development involve temporal and spatial specification and segregation of three late blastocyst cell lineages; trophectoderm (TE), primitive endoderm (PrE) and epiblast (EPI). Spatial separation of the outer TE lineage from the two inner-cell-mass (ICM) lineages (PrE and EPI) starts with the 8- to 16-cell transition and concludes following transit through the 16- to 32-cell stages. This results in a nascent early blastocyst ICM derived from descendants of primary founding inner cells and a secondarily contributed population; of which subsequent relative EPI versus PrE potencies are subject to debate. We showed that generation of primary but not the secondary ICM populations is highly dependent on temporally discreet activation of the mammalian target of rapamycin (mTOR – specifically mTORC1) around 8-cell stage M-phase entry. Moreover, that this role is mediated via the regulated function of the 7-methylguanosine- (7mG) cap binding initiation complex (EIF4F) and potentiating the translation of a subset of key but otherwise intransigent mRNAs containing 5’ UTR terminal oligopyrimidine (TOP-) sequence motifs. To find out translation of which mRNAs is regulated by mTOR during 8- to 16-cell transition, we analysed proteomes of control and mTOR-inhibited embryos during this time window.

2023-08-18 | PXD039423 | Pride

Gata3 acts alongside Cdx2 to promote trophoblast gene expression downstream of Tead4 during mouse development

Project description:The first lineage decisions during mouse development lead to establishment of embryonic and extraembryonic tissues. The transcription factor Cdx2 plays a central role by repressing pluripotency genes, such as Oct4 and promoting trophoblast fate at the blastocyst stage. Here we show that the transcription factor Gata3 is coexpressed with Cdx2 in the blastocyst and that overexpression of Gata3 in embryonic stem cells is sufficient to induce expression of trophoblast genes. Gata3 expression in the blastocyst does not depend on Cdx2, nor do Gata3 overexpressing cell lines require Cdx2 for expression of a subset of trophoblast genes. In the embryo, expression of Gata3, like Cdx2, depends on Tead4, and expression of both factors becomes restricted to nascent trophoblast by an Oct4-independent mechanism. These observations place Tead4 at the top of a trophoblast hierarchy, with Gata3 and Cdx2 acting downstream to induce expression of common and independent targets in this lineage. This SuperSeries is composed of the following subset Series: GSE12985: Differentiation time course of trophoblast stem cells GSE12986: Expression of Cdx2 or Gata3 in R1 mouse embryonic stem cells

2010-02-21 | E-GEOD-12999 | biostudies-arrayexpress

Single-cell analysis of gene networks in trophoectoderm and inner cell mass during human, monkey and mouse preimplantation development

Project description:Mammalian preimplantation development involves highly conserved genetic programs across species. However, temporal differences in gene network activation and membership differences in gene modules are also observed. In this study, we focus on understanding the activation of crucial gene networks in trophoectoderm (TE) and inner cell mass across species using human, monkey, and mouse models. By single cell RNA sequencing analysis in cynomolgus monkey early embryos, we find relatively late major embryonic genome activation occurring at early morula stages. Gene network preservation analysis reveals that genetic programs in monkey embryos more closely resemble human embryos compared to mouse, supporting the notion that preimplantation development in primate species are divergent from rodent species. For example, primate blastocysts show differential expression of ICM and TE genes such as X and Y, whereas mouse show differences in A and B expression. Interestingly, the overall conservation of TE gene networks is higher than ICM across three species, suggesting a more conserved developmental program in TE lineage differentiation. Additionally, TE-specific gene networks are exclusively activated at the blastocyst stage with low or no expression in morula or earlier embryos. In contrast, a majority of ICM-specific genes are strongly activated during the first major wave of EGA, which are further fine-tuned during late morula and blastocyst stages. Taken together, our study provides new insight into mammalian preimplantation development and sequential activation of lineage-specific genetic programs of TE versus ICM.

2022-06-30 | GSE75764 | GEO

Zona pellucida removal affects pre- and post-implantation development and gene expression in mouse embryos

Project description:The zona pellucida plays a crucial role in the process of fertilization to early embryonic development, including cellular arrangement and communication between blastomeres. However, little is known regarding the role of the zona pellucida in pre- and post-implantation embryonic development associated with gene expression. We investigated the effects of the zona pellucida on the development and gene expression of pre- and post-implantation embryos. After the zona pellucida was removed at the 2-cell stage, compaction occurred earlier. In addition, the expression of differentiation-related genes in the inner cell mass (ICM) (Oct4, Sox21) and trophectoderm (TE) (Cdx2, Eomes, and Tfap2c) was significantly altered in both zona free (ZF) morula and blastocyst compared to zona intact embryos. After embryo transfer, the implantation rate and number of live fetuses were lower in ZF embryos than in control embryos, whereas the fetal weight at birth was not different. However, placental weight was significantly increased in ZF embryos. RNA-seq analysis of the placenta showed that a total of 473 differentially expressed genes that significantly influenced the biological process. The present study suggests that zona pellucida removal at the 2-cell stage not only disturbs the expression pattern of ICM/TE-related genes, but affects the post-implantation development of mouse embryos. Overall, this study provides a deeper insight into the role of the zona pellucida during early mouse embryonic development and the viability of post-implantation development. It also provides a valuable basis for future research related to the production of ZF embryos in mammals and the developmental properties of blastomeres.

2022-03-31 | GSE199670 | GEO

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