Unknown,Transcriptomics,Genomics,Proteomics

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Early lineage restriction and regional segregation during mammalian heart development

ABSTRACT: 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.

ORGANISM(S): Mus musculus

SUBMITTER: Samira Chabab

PROVIDER: E-GEOD-59033 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Mouse heart development arises fromMesp1 expressing cardiovascular progenitors that are specified at the early stage of gastrulation. Lineage tracing and clonal analysisof Mesp1 progenitors revealed that heart development arises from distinct populations of Mesp1+ cardiovascular progenitors (CPs) expressing Mesp1 at different time points during gastrulation that contribute to different heart regions and different cardiovascular lineages. However, it remains unclear what are the molecular mechanisms that control the early regional and lineage segregation of Mesp1 CPs. Here, we performedsingle cell RNA-sequencing of FACS isolated Mesp1 CPs in WT and Mesp1 null embryos at different times to define the cellular and molecular heterogeneity of the Mesp1CPs, identify the role of Mesp1 in regulating cellular heterogeneity and uncover the mechanisms associated with lineage and regional segregation during the early stages of gastrulation.We showed that Mesp1 CPs isolated at E6.75 and E7.25 are molecularly distinct and make the continuum betweenepiblast and later mesodermal cells including hematopoietic progenitors. Single cell transcriptome of Mesp1 deficient CPsshowed that Mesp1 is required for the exitof thepluripotent state and the induction of the cardiovascular gene expression program in vivo.Using dimensional reduction analysis, we identified distinct populations of Mesp1 CPs that correspond to progenitors committed to different celllineages and regions of the heart, identifying the molecular features associated with the early lineage and regional segregation. Notch1CREER lineage tracing, a marker preferentially expressed by one of the different Mesp1CP subpopulations, during the early stage of gastrulation marked almost exclusively ECs, demonstrating theexistence of an early Mesp1 subpopulation committed to the EC fate.This study uncoversthe cellular and molecular heterogeneity associated with early lineage restriction and regional segregation of the heart at the early stage of gastrulation.

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Early lineage restriction and regional segregation during mammalian heart development

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