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Single-cell transcriptomics reveals chemotaxis mediated intra-organ crosstalk during cardiogenesis [ChIP-seq]

ABSTRACT: We used single-cell RNA-seq to reconstruct differentiation paths of cardiac progenitors in two sequential waves during early heart development. Further analysis identified six major cell types and multiple differentiation trajectories of cardiac progenitors derived from distinct heart fields. We also constructed TF regulatory networks controlling SHF CPC differentiation. Interlineage crosstalk through signaling pathways and chemotactic guidance played a potent role in SHF CPC deployment. The mechanisms regulating SHF CPC migration and differentiation was further confirmed by Nkx2-5 enhancer knock out. Our work provides a cardiac lineage hierarchy and new insights into SHF CPC development. Mechanistically, NKX2-5 directly bound the Cxcr2 genomic locus and activated its transcription.

ORGANISM(S): Mus musculus

PROVIDER: GSE108961 | GEO | 2019/06/22

REPOSITORIES: GEO

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Single-cell transcriptomic analysis reveals lineage hierarcies and communications for second heart lineage deployment [RNA-seq]

Project description:We used single-cell RNA-seq to reconstruct differentiation paths of cardiac progenitors in two sequential waves during early heart development. Further analysis identified six major cell types and multiple differentiation trajectories of cardiac progenitors derived from distinct heart fields. We also constructed TF regulatory networks controlling SHF CPC differentiation. Interlineage crosstalk through signaling pathways and chemotactic guidance played a potent role in SHF CPC deployment. The mechanisms regulating SHF CPC migration and differentiation was further confirmed by Nkx2-5 CPC enhancer knock out. Our work provides a cardiac lineage hierarchy and new insights of SHF CPC development.

2019-06-22 | GSE108879 | GEO

Expression analysis of genes regulated by RBPJ (Notch) and Axin2 (Wnt) mediated signaling in mouse cardiogenesis at E9.25

Project description:Progenitor cells of the first and second heart fields (FHF and SHF) depend on cardiac-specific transcription factors for their differentiation. In mouse mutant embryos, we define the hierarchy of signaling events that controls the expression of cardiac-specific transcription factors during commitment of SHF progenitors at E9.25. Wnt and Bmp act downstream of Notch/RBPJ at this developmental stage. Mutation of Axin2, the negative regulator of canonical Wnt signaling, enhances Wnt and Bmp signals and suffices to rescue the cardiac differentiation arrest caused by loss of RBPJ. By analysis of isolated cardiac progenitors, embryo cultures in the presence of pharmacological inhibitors, and Bmp triple mutants, we could classify the expression of heart-specific transcription factors of SHF progenitors according to their dependence on either Wnt or Bmp signals, Nkx2-5, Isl1, Baf60c and Gata4, SRF, Mef2c, respectively. Total RNA from whole embryonic hearts of control mice was compared to MesP1-cre:RBPJlox/lox (KO), MesP1-cre:RBPJlox/lox//Axin2-/- (DKO), MesP1-cre:RBPJlox/+//Axin2-/- (hetDKO) and MesP1-cre:RBPJlox/lox//Axin2+/- (DKOhet) mutant mouse embryos.

2012-06-20 | E-GEOD-36804 | biostudies-arrayexpress

Expression analysis of genes regulated by RBPJ (Notch) and Axin2 (Wnt) mediated signaling in mouse cardiogenesis at E9.25

2012-06-21 | GSE36804 | GEO

Transcription profiling of mouse second field myocardium Gata4flox/+ Nkx2-5+/+ (control, n=3), Gata4flox/+ Nkx2-5Cre/+ (Gata4 Nkx2-5 double heterozygous animals reveals GATA4 is a direct activator of Cyclin D2 and is required for proliferation in 2nd heart field-derived myocardium

Project description:The second heart field (SHF) comprises a population of mesodermal progenitor cells that are added to the nascent linear heart to give rise to the majority of the right ventricle, interventricular septum, and outflow tract of mammals and birds. The zinc finger transcription factor GATA4 functions as an integral member of the cardiac transcription factor network in the SHF and its derivatives. In addition to its role in cardiac differentiation, GATA4 is also required for cardiomyocyte replication, although the transcriptional targets of GATA4 required for proliferation have not been previously identified. In the present study, we disrupted Gata4 function exclusively in the SHF and its derivatives. Gata4 SHF knockout mice die by embryonic day 13.5 and exhibit hypoplasia of the right ventricular myocardium and interventricular septum and display profound ventricular septal defects. Loss of Gata4 function in the SHF results in decreased myocyte proliferation in the right ventricle, and we identify numerous cell cycle genes that are dependent on Gata4 by microarray analysis. We show that Gata4 is required for Cyclin D2 expression in the right ventricle and that the Cyclin D2 promoter is bound and activated by GATA4 via three consensus GATA binding sites. These findings establish Cyclin D2 as a direct transcriptional target of GATA4 and support a model in which GATA4 controls cardiomyocyte proliferation by coordinately regulating numerous cell cycle genes. Experiment Overall Design: Onewat ANOVA with post-hoc was done with 3 genotypes with replicates. Three genetypes are Gata4flox/+; Nkx2-5+/+ (control, n=3), Gata4flox/+; Nkx2-5Cre/+ (Gata4; Nkx2-5 double heterozygous, Experiment Overall Design: 170 n=3), and Gata4flox/flox; Nkx2-5Cre/+ (Gata4 CKONkx, n=5).

2008-10-20 | E-GEOD-9652 | biostudies-arrayexpress

Early lineage restriction and regional segregation during mammalian heart development

Project description:Cardiac development arises from two sources of mesoderm progenitors, the first (FHF) and the second heart field (SHF). Mesp1 has been proposed to mark the most primitive multipotent cardiac progenitors common for both heart fields. Here, using clonal analysis of the earliest prospective cardiovascular progenitors in a temporally controlled manner during the early gastrulation, we found that Mesp1 progenitors consist of two temporally distinct pools of progenitors restricted to either the FHF or the SHF. FHF progenitors were unipotent, while SHF progenitors, were either uni- or bipotent. Microarray and single cell RT-PCR analysis of Mesp1 progenitors revealed the existence of molecularly distinct populations of Mesp1 progenitors, consistent with their lineage and regional contribution. Altogether, these results provide evidence that heart development arises from distinct populations of unipotent and bipotent cardiac progenitors that independently express Mesp1 at different time points during their specification, revealing that the regional segregation and lineage restriction of cardiac progenitors occurs very early during gastrulation. We used microarrays to characterize the molecular mechanisms that control Mesp1 progenitor specification and lineage segregation during the early stage of cardiac mesoderm formation, 50 Mesp1 H2B-GFP+ or Mesp1 H2B-GFP- cells at E6.5 or E7.5 from mouse embryos were sorted for RNA extraction, amplification and hybridization on Affimetrix microarrays. Microaarrays were performed on Mouse Genome 430 PM strip Affymetrix array. The overall design was repeated in two different biological samples.

2014-07-10 | E-GEOD-59033 | biostudies-arrayexpress

Early lineage restriction and regional segregation during mammalian heart development

2014-07-10 | GSE59033 | GEO

Transcription profiling of mouse embryonic stem cell derived cardiac progenitors

Project description:Mamamlian cardiogenesis occurs through the development of discreate populations of first and second heart field progenitors. We have used a dual transgenic color reproter system to isolate purified populations of these progenitors. We used microarrays to detail the global programme of gene expression underlying cardiac development in mouse; All four populations of cells are derived from embryonic stem cells differentiating in vitro (day 6 of in vitro differentaition). The stem cell line has two transgenic reporters as follows:; 1. The second heart field (SHF) specific reporter of the Mef2C gene (E. Dodou, S. M. Xu, B. L. Black, Mech Dev 120, 1021 (Sep, 2003)) driving the expression of dsRed; 2. The cardiac specific enhancer ( C. L. Lien et al., Development 126, 75 (Jan, 1999)) driving the expression of eGFP. Thus, the red cells are SHF specific, the green cells are cardiac specific, and the red/green are SHF and cardiac specific. These cells are compared to the double negative cells which serve as a control. Experiment Overall Design: Embryonic stem cell derived progenitors were isolated into four distinct populations by FACS purifying these progenitors based on a two color reporter system. Four populations were then compared to each other by transcriptional profiling.

2009-09-12 | E-GEOD-17513 | biostudies-arrayexpress

Complex Interdependence Regulates Heterotypic Transcription Factor Distribution and Coordinates Cardiogenesis [Chip-Seq]

Project description:Genome-wide occupancy analysis of TBX5, NKX2-5 and GATA4 in differentiating WT, Nkx2-5KO (NKO), Tbx5KO (TKO) and Nkx2-5;Tbx5KO (DKO) cells at the cardiac precursor (CP) and cardiomyocyte (CM) differentiation stages. Analysis of TBX5, NKX2-5 and GATA4 occupancy a and gene expression in WT, Tbx5KO, Nkx2-5KO and DoubleKO precursor (CP) and mature (CM) in vitro differentiated cardiomyocytes.

2016-02-11 | GSE77548 | GEO

Expression data from murine embryonic stem cell derived cardiac progenitors

Project description:Mamamlian cardiogenesis occurs through the development of discreate populations of first and second heart field progenitors. We have used a dual transgenic color reproter system to isolate purified populations of these progenitors. We used microarrays to detail the global programme of gene expression underlying cardiac development in mouse All four populations of cells are derived from embryonic stem cells differentiating in vitro (day 6 of in vitro differentaition). The stem cell line has two transgenic reporters as follows: 1. The second heart field (SHF) specific reporter of the Mef2C gene (E. Dodou, S. M. Xu, B. L. Black, Mech Dev 120, 1021 (Sep, 2003)) driving the expression of dsRed 2. The cardiac specific enhancer ( C. L. Lien et al., Development 126, 75 (Jan, 1999)) driving the expression of eGFP. Thus, the red cells are SHF specific, the green cells are cardiac specific, and the red/green are SHF and cardiac specific. These cells are compared to the double negative cells which serve as a control.

2009-09-01 | GSE17513 | GEO

Complex Interdependence Regulates Heterotypic Transcription Factor Distribution and Coordinates Cardiogenesis [microarray]

Project description:In the developing heart, heterotypic transcription factors (TFs) interactions, such as between the T-box TF TBX5 and the homeodomain TF NKX2-5 have been proposed as a mechanism for human congenital heart disease. In order to study the role of each TF during heart formation, embryonic stem (ES) cell-derived embryos were generated from KO ES cells for Tbx5, Nkx2-5 or both TFs. We used microarrays to identify changes in the gene expression due to the lack of Tbx5, Nkx2-5 or both TFs during early heart formation. WT, Nkx2-5KO (NKO), Tbx5KO (TKO) and Tbx5KO;Nkx2-5KO (DKO) E8.75 mouse hearts were microdissected for RNA extraction and hybridization on Affymetrix microarrays.

2016-02-11 | E-GEOD-77576 | biostudies-arrayexpress

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