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Gene expression analysis of control and Tbx20 mutant Tie2Cre lineage from E12.5 hearts

ABSTRACT: To investigate roles for Tbx20 in endocardium, we ablated Tbx20 utilizing Tie2Cre. Tie2Cre;Tbx20 mutants died at E14, exhibiting defects in multiple aspects of cardiac septation. Although endocardial cells lacking Tbx20 were able to undergo endothelial-to-mesenchymal transition, cushion mesenchymal cells lacking Tbx20 did not disperse normally. Non-cell autonomous roles of endocardial Tbx20 were also revealed, as evidenced by decreased myocardialization of outflow tract and failure of dorsal mesenchymal protrusion formation in mutants. To examine how ablation of Tbx20 in endocardial lineages affected gene expression, we performed global gene expression analysis on purified endocardial lineages. E12.5 hearts were dissociated, and Tie2Cre;RosatdTom lineage traced cells of controls and mutants were isolated by fluorescence activated cell sorting (FACS), after exclusion of blood cells (Ter119+, CD41+ and/or CD45+). Mutant endocardial lineages exhibited decreased expression of genes associated with extracellular matrix and cell migration. E12.5 hearts were dissociated, and Tie2Cre;RosatdTom lineage traced cells of controls and mutants were isolated by fluorescence activated cell sorting (FACS), after exclusion of blood cells (Ter119+, CD41+ and/or CD45+).

ORGANISM(S): Mus musculus

PROVIDER: GSE73858 | GEO | 2016/07/01

SECONDARY ACCESSION(S): PRJNA298232

REPOSITORIES: GEO

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Gene expression analysis of control and Tbx20 mutant Tie2Cre lineage from E12.5 hearts

Project description:To investigate roles for Tbx20 in endocardium, we ablated Tbx20 utilizing Tie2Cre. Tie2Cre;Tbx20 mutants died at E14, exhibiting defects in multiple aspects of cardiac septation. Although endocardial cells lacking Tbx20 were able to undergo endothelial-to-mesenchymal transition, cushion mesenchymal cells lacking Tbx20 did not disperse normally. Non-cell autonomous roles of endocardial Tbx20 were also revealed, as evidenced by decreased myocardialization of outflow tract and failure of dorsal mesenchymal protrusion formation in mutants. To examine how ablation of Tbx20 in endocardial lineages affected gene expression, we performed global gene expression analysis on purified endocardial lineages. E12.5 hearts were dissociated, and Tie2Cre;RosatdTom lineage traced cells of controls and mutants were isolated by fluorescence activated cell sorting (FACS), after exclusion of blood cells (Ter119+, CD41+ and/or CD45+). Mutant endocardial lineages exhibited decreased expression of genes associated with extracellular matrix and cell migration. E12.5 hearts were dissociated, and Tie2Cre;RosatdTom lineage traced cells of controls and mutants were isolated by fluorescence activated cell sorting (FACS), after exclusion of blood cells (Ter119+, CD41+ and/or CD45+). FACS sorted Tie2Cre lineage from E12.5 hearts: Tie2Cre;Tbx20 +/loxP Control hearts versus Tie2Cre;Tbx20 loxP/- mutant hearts

2016-07-01 | E-GEOD-73858 | biostudies-arrayexpress

Expression data from murine cardiac fibroblasts and endothelial cells following Transverse Aortic Constriction

Project description:Accumulation of activated cardiac fibroblasts plays a key role in heart failure progression. These cells deposit excessive extracellular matrix that leads to mechanical stiffness, myocyte uncoupling and ischemia. To investigate whether two developmentally distinct cardiac fibroblast populations exhibit distinct expression profiles in response to cardiac injury, and therefore might necessitate distinct therapeutic targeting, we performed microarray analysis on FACS sorted cells. Tie2cre lineage traced CFs, non Tie2cre lineage traced cardiac fibroblasts and endothelial cells were isolated from left ventricle of SHAM operated and banded hearts at the onset of fibrosis, one week after surgery. We used microarrays to detail the global programme of gene expression in cardiac fibroblasts and endothelium following pressure overload. Tie2cre lineage traced, non-tie2cre lineage traced fibroblasts and endothelial cells were sorted from left ventricle of 3 SHAM operated and 3 TAC operated adult male Black Swiss mice. Tie2cre lineage traced, non-tie2cre lineage traced fibroblasts and endothelial cells were sorted from left ventricle of 3 SHAM operated and 3 Transaortic Constriction (TAC) operated adult male Black Swiss mice for RNA extraction and Affymetrix microarray analysis. Hypertrophy in TAC animals, an lack of hypertrophy in SHAM operated animals, was evaluated by hemodynamic measurements before surgery and one week after surgery. Cells were isolated one week after surgery.

2014-05-19 | E-GEOD-45820 | biostudies-arrayexpress

Mus musculus

Project description:Gene expression analysis of control and Tbx20 mutant Tie2Cre lineage from E12.5 hearts

| PRJNA298232 | ENA

Expression data from murine cardiac fibroblasts and endothelial cells following Transverse Aortic Constriction

2014-05-19 | GSE45820 | GEO

ChIP-seq of Tbx20 tagged with GFP in E11.5 mouse hearts

Project description:We performed ChIP-seq of Tbx20 tagged with GFP in mouse hearts to identify binding sites for this transcription factor. This data set is part of the study âEndocardial Tbx20 is essential for mesenchymal and myocardial cell movements required for cardiac septationâ.

2016-06-30 | E-MTAB-3967 | biostudies-arrayexpress

ATAC-seq on endocardial cells from E12.5 mouse hearts in Tbx20 knockouts and wild type counterparts

Project description:We used ATAC-seq (Buenrostro et al, 2013) to identify regions of open chromatin in FACS sorted mouse endothelial cells from E12.5 hearts. Please note that the animals were injected at different dates, which resulted in different effects of the knockout, as indicated in the batch attribute of the samples. This data set is part of the study \Endocardial Tbx20 is essential for mesenchymal and myocardial cell movements required for cardiac septation\. Peaks were called using HOMER (-gsize 1.87e9 -region -tbp 1). Replicate 1: -fdr 1e-8, replicate 2: default parameters. Buenrostro JD, Giresi PG, Zaba LC, Chang HY, Greenleaf WJ. 2013. Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position. Nat Methods 10: 12138.

2017-06-02 | E-MTAB-3972 | biostudies-arrayexpress

Lineage selective cardiac GATA4 chromatin occupancy [embryo]

Project description:We used cre-directed biotinylation of a GATA4 knockin allele with an AviTag epitope tag to measure GATA4 chromatin occupancy in endocardial and cardiomyocyte lineages in the E12.5 heart. These data are complemented by lineage selective measurement of gene expression and chromatin accessibility. Cardiomyocyte accessibility and gene expression data were reported in GSE124008.

2021-07-01 | GSE156001 | GEO

Microarray using CD31+/CD41-/CD45- cells from E9.5 mouse heart tube, caudal half and yolk sac

Project description:Hematopoietic cells arise from spatiotemporally restricted domains in the developing embryo. Although studies of non-mammalian animal and in vitro embryonic stem cell models suggest a close relationship among cardiac, endocardial, and hematopoietic lineages, it remains unknown whether the mammalian heart tube serves as a hemogenic organ akin to the dorsal aorta. Here, we examined the hemogenic activity of the developing endocardium. Mouse heart explants generated myeloid and erythroid colonies in the absence of circulation. Hemogenic activity arose from a subset of endocardial cells in the outflow cushion and atria earlier than in the aorta-gonad-mesonephros region, and was transient and definitive in nature. Interestingly, key cardiac transcription factors, Nkx2-5 and Isl1, were expressed in and required for the hemogenic activity of the endocardium. Together, these data suggest that a subset of endocardial and yolk sac endothelial cells expressing cardiac markers serve as a de novo source for transient definitive hematopoietic progenitors. Two independent biological duplicates of freshly isolated mouse tissues (caudal half, heart tube, yolk sac) were sorted for CD31+/CD41-/CD45- cells.

2013-08-17 | E-GEOD-40260 | biostudies-arrayexpress

Indexed single-cell RNA-seq of mensenchymal bone marrow cells

Project description:This data set contains single-cell RNA-seq data from CD45-Ter119-Cd41-CD71-Vibrant Dye+VCAM1+ cells, indexed for the expression of these markers as well as CD51, CD61 and CD200

2019-08-08 | GSE134098 | GEO

Single cell RNA-seq of mouse megakaryocytes from wild type and Nf-e2 knock out mice

Project description:Fetal livers from 8X E13.5 wildtype and 6X Nf2-/- mutant embryos were mechanically dissociated, pooled by genotype and stained for CD41, CD42d, LIN (Ter119,B220). LIN- CD41high CD42d+ Cells were sorted on an Aria Cell Sorter (Becton Dickinson), centrifuged and counted. Fetal livers of 5X E13 wildtype or Nf2-/- embryos were prepared as above and transplanted into UBC-GFP+ recipients subjected to 2X 550cGrays irradiations (0.37 embryonic equivalent/recipient). Mice were screened at 4 weeks for blood platelet content and their GFP status. Bone marrow from the femurs, tibias, hips and humeri of 4 transplanted mice (2X WT donor, 2X Nf2-/- donor) was dissociated, pooled by genotype, filtered and stained for CD41, CD42d, LIN (Ter119, B220). GFP- LIN- CD41high CD42d+ Cells were sorted on an Aria Cell Sorter (Becton Dickinson), centrifuged and counted.

2020-07-15 | GSE125579 | GEO

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