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ChIP-on-chip with anti-RBPj antibody from embryonic AGM E11.5

ABSTRACT: Hematopoietic Stem Cells (HSC) are originated during embryonic development from endothelial-like cells located in the ventral side of the dorsal aorta around day E10-12 of murine development. This region is called AGM for Aorta/Gonad/Mesonephros refering to the tissues around the hemogenic aorta. Hematopoiesis depends on the Notch pathway and the identification of Notch-targets is important for the understanding of blood origin.

ORGANISM(S): Mus musculus

PROVIDER: GSE52094 | GEO | 2014/10/22

SECONDARY ACCESSION(S): PRJNA226668

REPOSITORIES: GEO

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ChIP-on-chip with anti-RBPj antibody from embryonic AGM E11.5

Project description:Hematopoietic Stem Cells (HSC) are originated during embryonic development from endothelial-like cells located in the ventral side of the dorsal aorta around day E10-12 of murine development. This region is called AGM for Aorta/Gonad/Mesonephros refering to the tissues around the hemogenic aorta. Hematopoiesis depends on the Notch pathway and the identification of Notch-targets is important for the understanding of blood origin. Hematopoietic Stem Cells (HSCs) specification occurs in the embryonic aorta and requires Notch activation, however which are the elements regulated by Notch that control this process are mainly unknown. Here, we took a genome-wide approach to identify putative direct Notch targets by precipitating the chromatin that binds to the Notch partner RBPj in the Aorta-Gonad-Mesonephros (AGM) tissue from E11.5 mouse embryos. This assay revealed 701 gene promoter regions as candidates to be regulated by Notch in the AGM. Chromatin was obtained from a pool of 40 dissected AGMs at E11.5. Chromatin immunoprecipitation (ChIP) was performed as previously described (Aguilera et al, PNAS 2004) with minor modifications. In brief, cross-linked chromatin was sonicated for 10 minutes, medium-power, 0.5-interval; with a Bioruptor (Diagenode) and precipitated with anti-RBPJ (Chu and Bresnick, 2004). After crosslinkage reversal, DNA was used as a template for PCR or for array hybridization. Mouse promoter chip on chip microarray SET (Agilent) was used to identify RBPj targets. It covers 70,000 best identified gene regions with a-5.5 kb to + 2.5 kb range, and has on average 25 probes per gene with an average probe to probe distance of 200 bp. The ChIP-on-chip was performed with dye swaps and one IgG control was brought along. Enrichment analysis was done by comparing the precipitation normalized dye swap signal with input control signal.

2014-10-22 | E-GEOD-52094 | biostudies-arrayexpress

Transcriptome imposed by Notch ligands (Jag1 or Dll4) on ckit+ and ckit- cells from E11.5 AGM

2015-08-20 | GSE59344 | GEO

Expression from early pre-hematopoietic progenitors from mouse embryo

2012-07-03 | GSE35395 | GEO

Expression from early pre-hematopoietic progenitors from mouse embryo

2012-07-02 | E-GEOD-35395 | biostudies-arrayexpress

Bulk RNA sequencing of CD44Neg, CD44LoKitNeg, CD44LoKitPos and CD44High populations from mouse Aorta-Gonad-Mesonephros (AGM) region

Project description:The aim of the experiment was to compare newly defined CD44Neg, CD44LoKitNeg, CD44LoKitPos and CD44High populations from mouse Aorta-Gonad-Mesonephros (AGM) region

2019-03-29 | GSE128971 | GEO

Expression data of endothelium and hematopoietic cluster cells in the wild type and Steel mutant E11.5 AGM and vitelline and umbilical arteries

Project description:Kit ligand (Kitl, aka SCF) is a pivotal cytokine in fetal liver and bone marrow hematopoiesis. Its role in pre-liver embryonic hematopoiesis, however, is less well understood. We investigated the hematopoietic phenotype of Steel (Sl/Sl) mutant embryos, which lack Kitl, and found that the normal development of the HSC lineage in the aorta-gonad-mesonephros (AGM) region is negatively affected. We isolated endothelium and hematopoietic cluster cells from wild type and Sl/Sl AGM and vitelline and umbilical arteries for expression analysis by RNA-seq to investigate molecular changes in the developing HSC lineage and endothelial cells in the aortic HSC niche.

2018-08-30 | GSE105267 | GEO

Engineering a Niche Supporting Hematopoietic Stem Cell Development Using Integrated Single Cell Transcriptomics

Project description:Hematopoietic stem cells (HSC) develop from hemogenic endothelium (HE) within embryonic arterial vessels such as the aorta of the aorta-gonad-mesonephros region (AGM). To identify the signals responsible for HSC formation, we used single cell RNA-sequencing to simultaneously analyze the transcriptional profiles of AGM-derived cells transitioning from HE to HSC, and AGM-derived endothelial cells which provide signals sufficient to support HSC maturation and self-renewal. Pseudotemporal ordering revealed dynamics of gene expression during the HE to HSC transition, identifying surface receptors specifically expressed on developing HSC. Transcriptional profiles of niche endothelial cells enabled identification of corresponding ligands, including those signaling to Notch receptors, VLA-4 integrin, and CXCR4, which, when integrated in an engineered platform, were sufficient to support the generation of engrafting HSC. These studies provide a transcriptional map of the signaling interactions necessary for the development of HSC and advance the goal of engineering HSC for therapeutic applications

2020-03-01 | GSE145886 | GEO

De novo generation of macrophage from placenta-derived hemogenic endothelium

Project description:We profile the transcriptional landscape of mouse fetal placenta and identify four distinct types of haematopoietic stem/progenitor cells (HSPCs) during mid-gestation using single-cell RNA sequencing. We experimentally validate and uncover that a subpopulation of placental endothelial cells exhibits hemogenic endothelial (HE) potential and shares transcriptional features with HE cells in the aorta-gonad-mesonephros (AGM) region.

2021-06-30 | GSE152903 | GEO

A molecular roadmap of the aorta-gonad-mesonephros region reveals BMPER as a novel regulator of HSC maturation [AGM]

Project description:In the developing embryo, haematopoietic stem cells (HSCs) emerge from the aorta-gonad-mesonephros (AGM) region but the molecular regulation of this process is poorly understood. Recently, the progression from E9.5 to E10.5 and polarity along the dorso-ventral axis have been identified as clear demarcations of the supportive HSC niche. To identify novel secreted regulators of HSC maturation, we performed RNA-sequencing over these spatio-temporal transitions in the AGM region, and supportive OP9 cell line.

2017-10-30 | GSE102857 | GEO

Spatiotemporal and functional heterogeneity of the hematopoietic stem cell-competent hemogenic endothelial cells in mouse embryos

Project description:Hematopoietic stem cells (HSCs) are derived from hemogenic endothelial cells (HECs) during embryogenesis. The HSC-primed HECs are peak at embryonic day (E) 10 and have been efficiently captured by the marker combination CD41-CD43-CD45-CD31+CD201+Kit+CD44+ (PK44) in the aorta-gonad-mesonephros (AGM) region of mouse embryos most recently. In the present study, we investigated the spatiotemporal and functional heterogeneity of PK44 cells around the time of emergence of HSCs. First, PK44 cells in E10 AGM region could be further divided into three molecularly different populations showing endothelial- or hematopoietic-biased characteristics. Specifically, with the combination of Kit, the expression of CD93 or CD146 could divide PK44 cells into endothelial- and hematopoietic-feature biased populations, which was further functionally validated at single cell level. Next, PK44 population could also be detected in the yolk sac, showing a developmental dynamics and functional diversification similar to those in the AGM region. Importantly, PK44 cells in the yolk sac demonstrated an unambiguous multi-lineage reconstitution capacity after in vitro incubation. Regardless of the functional similarity, PK44 cells in the yolk sac displayed transcriptional features different to those in the AGM region. Taken together, our work delineated the spatiotemporal characteristics of HECs represented by PK44, and revealed a previously unknown HSC competence of HECs in the yolk sac. These findings provided a fundamental basis for in-depth studying the different origins and molecular programs of HSC generation in the future.

2021-08-20 | GSE173833 | GEO

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