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Specific gene expression related to epiblast development

ABSTRACT: To explore genes involved in epiblast development, we have employed whole genome microarray expression profiling as a discovery platform to identify genes. Embryoid body (EB) formation induces embryonic stem (ES) cells to differentiate into germ layers via epiblasts and extra-embryonic tissues, whereas ES cells give rise only to extra-embryonic tissues in monolayer culture. We compared gene expression profiles of ES cells in each differentiation culture methods at day 1-3 or 4 of differentiation. The expression of organizer related genes (Lhx1, Cer1 and Foxa2) was specifically induced at day 3 of EBs, but not in monolayer culture. In epiblast model cells, the overexpression of Lhx1 induced gene expression of organizer and meso-endodermal markers, suggested that Lhx1 was related to epiblast differentiation. Gene expression related epiblast development was measured at undifferentiated state and day 1-3 or 4 of differentiation. Two independent experiments were performed at each time using different people for each experiment.

ORGANISM(S): Mus musculus

SUBMITTER: Akira Hasegawa

PROVIDER: E-GEOD-64715 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Specific gene expression related to epiblast development

Project description:To explore genes involved in epiblast development, we have employed whole genome microarray expression profiling as a discovery platform to identify genes. Embryoid body (EB) formation induces embryonic stem (ES) cells to differentiate into germ layers via epiblasts and extra-embryonic tissues, whereas ES cells give rise only to extra-embryonic tissues in monolayer culture. We compared gene expression profiles of ES cells in each differentiation culture methods at day 1-3 or 4 of differentiation. The expression of organizer related genes (Lhx1, Cer1 and Foxa2) was specifically induced at day 3 of EBs, but not in monolayer culture. In epiblast model cells, the overexpression of Lhx1 induced gene expression of organizer and meso-endodermal markers, suggested that Lhx1 was related to epiblast differentiation.

2015-04-01 | GSE64715 | GEO

Dynamic proteomic profiling of extra-embryonic endoderm differentiation in mouse embryonic stem cells

Project description:During mammalian pre-implantation development, the cells of the blastocyst’s inner cell mass differentiate into the epiblast and primitive endoderm lineages, which give rise to the fetus and extra-embryonic tissues, respectively. Extra-embryonic endoderm differentiation can be modeled in vitro by induced expression of GATA transcription factors in mouse embryonic stem cells. Here we use this GATA-inducible system to quantitatively monitor the dynamics of global proteomic changes during the early stages of this differentiation event and also investigate the fully differentiated phenotype, as represented by embryo-derived extra-embryonic endoderm (XEN) cells. Using mass spectrometry-based quantitative proteomic profiling with multivariate data analysis tools, we reproducibly quantified 2,336 proteins across three biological replicates and have identified clusters of proteins characterized by distinct, dynamic temporal abundance profiles. We first used this approach to highlight novel marker candidates of the pluripotent state and extra-embryonic endoderm differentiation. Through functional annotation enrichment analysis, we have shown that the downregulation of chromatin-modifying enzymes, the re-organization of membrane trafficking machinery and the breakdown of cell-cell adhesion are successive steps of the extra-embryonic differentiation process. Thus, applying a range of sophisticated clustering approaches to a time-resolved proteomic dataset has allowed the elucidation of complex biological processes which characterize stem cell differentiation and could establish a general paradigm for the investigation of these processes.

2015-06-13 | PXD001901 | Pride

Neural induction (N2B27 monolayer cluture)

Project description:Neural induction of ES by N2B27 monolayer culture Keywords: development or differentiation design,time series design

2006-01-23 | GSE4076 | GEO

Occupancies of tissue-specific cis-regulatory modules by Spemann-Mangold organizer-specific transcription factors for embryonic head specification

Project description:Spemann-Mangold organizer-specific transcription factors (TFs), Otx2, Lim1/Lhx1, and Gsc, are essential for embryonic head specification. However, the molecular mechanisms by which those TFs interact with the genome to direct head formation are largely unknown. Here we employed ChIP-seq and RNA-seq approaches in Xenopus tropicalis gastrulae and found that occupancy of the corepressor, TLE/Groucho, is a better indicator of tissue-specific cis-regulatory modules (CRMs) than the coactivator p300, during early embryonic stages. On the basis of TLE binding and comprehensive CRM profiling, we defined two distinct types of Otx2- and TLE-occupied CRMs. Using these devices, Otx2 and Lim1/Lhx1 (activator) or Goosecoid (repressor) are able to upregulate or downregulate a large battery of target genes in the head organizer. An underlying principle is that Otx marks target genes for head specification to be regulated positively or negatively by partner TFs through specific types of CRMs.

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Neural induction (N2B27 monolayer cluture)

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2006-01-22 | E-GEOD-4076 | biostudies-arrayexpress

Lhx1 is required for specification of renal progenitor cell field

Project description:The goal of this study is to investigate the molecular mechanism of lhx1 on regulation of pronephros formation during the early embryonic development. In the vertebrate embryo the kidney is derived from the intermediate mesoderm. The LIM-class homeobox transcription factor lhx1 is expressed early in the intermediate mesoderm and is one of the first genes to be expressed in the nephric mesenchyme. The animal cap cells can be induced by treatment of activin and retinoic acid to differentiate into pronephros tissue. In this study we investigated the role of Lhx1 in differentiation of pronephros by depleting lhx1 in the organ culture system. We generated the gene expression profile of early pronephros tissue, and demonstrated that expression of genes from all the kidney domains is affected by the absence of lhx1. Taken together our results highlight an essential role for Lhx1 in pronephros formation.

2010-09-29 | GSE24392 | GEO

Differentiation to extra-embryonic primitive endoderm from naïve human ESCs on mouse embryonic fibroblasts (MEFs) or laminin-511 (LN511) substrates

Project description:Embryonic stem cells (ESCs) can exist in at least two states that transcriptionally resemble different stages of embryonic development. Naïve ESCs resemble peri-implantation stages and primed ESCs the pre-gastrulation epiblast. In mouse, primed ESCs give rise to definitive endoderm in response to pathways downstream of Nodal and Wnt signalling. However, when these cytokines are applied to naïve ESCs, they differentiate to a cell type that approximates early primitive endoderm (PrE), the blastocyst stage progenitor layer that gives rise to the extra-embryonic endoderm. Here, we apply this context dependency to human ESCs, showing that these cytokines drive the differentiation of naïve pluripotent cells to generate extra-embryonic PrE, or hypoblast, and, as in mouse, expanded as an in vitro model for naïve extra-embryonic endoderm (nEnd) in defined conditions. Consistent with observations made in mouse, human PrE differentiation is dependent on FGF signalling in vitro, and we show, that by inhibiting FGF receptor signalling, we could simplify naïve pluripotent culture such that inhibitor requirements closer resembled those used in mouse. These nEnd cultures represent stable extra-embryonic endoderm, or human hypoblast, cell lines.

2019-11-30 | GSE138013 | GEO

Naïve human pluripotency stem cells respond to Wnt, Nodal and LIF signalling to produce expandable naïve extra-embryonic endoderm

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Novel cell lines from mouse epiblast share defining features with human embryonic stem cells

Project description:The application of human embryonic stem (ES) cells has an inherent reliance on understanding the starting cell population. Human ES cells differ from mouse ES cells and the specific embryonic origin of both cell types is unclear. Previous work suggested that mouse ES cells could only be obtained from the embryo prior to implantation in the uterus. Here we show that cell lines can be derived from the epiblast, a tissue of the post-implantation embryo that generates the embryo proper. These cells, which we refer to as EpiSCs (post-implantation epiblast-derived stem cells), express transcription factors known to regulate pluripotency, maintain their genomic integrity, and robustly differentiate into the major somatic cell types as well as primordial germ cells (PGCs). The post-ES cell lines are distinct from mouse ES cells in their epigenetic state and the signals controlling their differentiation. Furthermore, post-ES and human ES cells share patterns of gene expression and signalling responses that normally function in the epiblast. These results show that epiblast cells can be maintained as stable cell lines and interrogated to understand how pluripotent cells generate distinct fates during early development. This SuperSeries is composed of the SubSeries listed below.

2007-06-27 | GSE7902 | GEO

RISC-mediated control of selected chromatin regulators stabilizes ground state pluripotency of mouse embryonic stem cells (RNA)

Project description:We performed an integrated analysis of RNA and proteins at the transition between naÃ¯ve ES cells and cells primed to differentiate. During this transition, mRNAs coding for chromatin regulators were specifically released from translational inhibition mediated by RNA-Induced Silencing Complex (RISC). This suggests that, prior to differentiation, the propensity of ES cells to change their epigenetic status is hampered by RNA interference. The expression of these chromatin regulators was reinstated following acute inactivation of RISC, and it correlated with loss of stemness markers and activation of early cell differentiation markers in treated ES cells. Comparison between EpiSC derived directly from mouse embryonic stem cells and from Epiblast-Like Aggregates

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