Mus musculus
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA311733 | ENA |
REPOSITORIES: ENA
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Project description:Gap junctional intercellular communication (GJIC) has been suggested to be involved in early embryonic development but the actual functional role remained elusive. Connexin (Cx) 43 and Cx45 are co-expressed in embryonic stem (ES) cells, form gap junctions and are considered to exhibit adhesive function and/or to contribute to the establishment of defined communication compartments. Here we describe the generation of Cx43/Cx45-double deficient mouse ES cells to achieve almost complete breakdown of GJIC. Cre-loxP induced deletion of both, Cx43 and Cx45, results in a block of differentiation in embryoid bodies without affecting pluripotency marker expression and proliferation in ES cells. We demonstrate that GJIC-incompetent ES cells fail to form primitive endoderm in embryoid body cultures, representing the inductive key step of further differentiation events in this system. Lentiviral overexpression of either Cx43 or Cx45 in Cx43/45 mutants rescued the observed phenotype, confirming the specificity and indicating a partially redundant function of both connexins. Upon differentiation GJIC-incompetent ES cells exhibit a strikingly altered subcellular localization pattern of the transcription factor NFATc3. Control EBs exhibit significantly more activated NFATc3 in cellular nuclei than mutant EBs suggesting that Cx-mediated communication is needed for synchronized NFAT activation to induce orchestrated primitive endoderm formation. Moreover, pharmacological inhibition of NFATc3 activation by Cyclosporin A, a well-described inhibitor of calcineurin, phenocopies the loss of GJIC in control cells.
2016-11-22 | GSE77852 | GEO
Project description:Analysis of embryonic sten cell-derived embryoid bodies following endoglin knock out. Loss of endoglin leads to profound reduction of key hematopoietic regulators including SCL, LMO2, Gata2, and TGF-? signaling molecule ALK-1. Results provide insight into molecular mechanisms underlying hemangioblast and primitive hematopoietic development. Total RNA obtained from differentiated day 3 EBs of endoglin knock out ES cells were compared to wild type E14 control ES cells.
2011-05-16 | E-GEOD-29300 | biostudies-arrayexpress
Project description:Establishment of left-right (LR) asymmetry occurs shortly after gastrulation and utilizes a cascade of events. In the mouse, LR symmetry is broken at the node, involves signal relay to the lateral plate, and results in asymmetric organ morphogenesis. How information transmits from the node to the lateral plate remains unclear. Noting that embryos lacking Sox17 exhibit defects in both gut endoderm formation and LR patterning, we investigated a connection between these two processes. We noted an endoderm-specific absence of the critical gap junction component, Connexin43, in Sox17 mutants. Dye-coupling experiments revealed planar gap junction coupling across the gut endoderm in wild-type but not mutant embryos. The role for gap junction communication in LR patterning was confirmed by pharmacological inhibition. Collectively, our data demonstrate communication across gap junctions in gut endoderm as a mechanism for information relay between node and lateral plate critical for the establishment of LR asymmetry in mice. Total RNA isolated from embryonic regions of wild-type embryos at EHF stages, three samples per well, in triplicates
2011-11-04 | E-GEOD-33353 | biostudies-arrayexpress
Project description:These data include the genome wide location of different histone modifications by ChIP sequencing in mouse ES cells, and RNA Seq data generated from wild type and EED KO mouse ES cells and knocked down for unrelated protein and Setd2 protein. ChIP-Seq: Immuno-precipitation of formaldehyde cross-linked chromatin prepared from wild type mouse E14 ES cells, wild type E36 ES cells, EED KO E36 ES cells, wild type Embryoid bodies (Ebs), EED KO Embryoid bodies (Ebs EED KO) using specific antibody against different histone modifications. RNA-Seq: Total RNA extracted from wild type E36 ES cells, EED KO E36 ES cells, wild type E36 Embryoid bodies (Ebs), EED KO Embryoid bodies (Ebs EED KO), E14 Ctrl KD, E14 Setd2 KD.
2013-12-05 | E-GEOD-51006 | biostudies-arrayexpress
Project description:Embryonic stem (ES) cells, when grown in suspension culture without feeders, spontaneously form round structures known as embryoid bodies. Given the appropriate conditions, these cells can differentiate over time into precursors of all three germ layers. Embryoid bodies, in a disorganized way, mimic early embryonic development to a certain extent and can be used as a synchronously differentiating large scale source of tissue for the study of biological determinants of early differentiation. Embryoid bodies have been used as a source for most early protocols that derive specific differentiated cell types from undifferentiated ES cells, although some protocols, notably those that derive neurons from ES cells, have moved on from EBs as a result of varying replicability and yield. We have decided to look at the transcriptomic profiles of embryoid bodies during the initial stages of embryoid body formation and differentiation, in order to pinpoint novel determinants of key developmental stages. Keywords: time course
2007-08-15 | GSE8766 | GEO
Project description:Embryoid Bodies from Mouse R1 ES - 9 day Keywords: other
2005-07-28 | GSE3007 | GEO
Project description:Mst1/Mst2 are central components of Hippo pathway. We examined the role of Mst1/Mst2 in ES cell differentiation. In this data set, we include expression data from day 4 and day 8 Mst1/Mst2 knockout ES cell formed embryoid bodies and wild type embryoid body controls.
2014-01-02 | GSE50219 | GEO
Project description:To identify potential Elongin A targets during neuronal differentiation of ES cells, a cDNA microarray analysis comparing embryoid bodies (EBs) derived from Elongin A+/+ ES cells and Elongin A-/- ES cells was performed.
2011-07-26 | GSE30911 | GEO