Article | Published:

Early lineage restriction in temporally distinct populations of Mesp1 progenitors during mammalian heart development

Nature Cell Biology volume 16, pages 829840 (2014) | Download Citation

Abstract

Cardiac development arises from two sources of mesoderm progenitors, the first heart field (FHF) and the second (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 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, whereas SHF progenitors were either unipotent or bipotent. Microarray and single-cell PCR with reverse transcription analysis of Mesp1 progenitors revealed the existence of molecularly distinct populations of Mesp1 progenitors, consistent with their lineage and regional contribution. Together, 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 occur very early during gastrulation.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

Gene Expression Omnibus

References

  1. 1.

    & A common progenitor at the heart of development. Cell 127, 1101–1104 (2006).

  2. 2.

    , & The arterial pole of the mouse heart forms from Fgf10-expressing cells in pharyngeal mesoderm. Dev. Cell 1, 435–440 (2001).

  3. 3.

    , , , & The clonal origin of myocardial cells in different regions of the embryonic mouse heart. Dev. Cell 6, 685–698 (2004).

  4. 4.

    et al. MesP1 is expressed in the heart precursor cells and required for the formation of a single heart tube. Development 126, 3437–3447 (1999).

  5. 5.

    & Mesp1: a key regulator of cardiovascular lineage commitment. Circ. Res. 107, 1414–1427 (2010).

  6. 6.

    et al. Mesp1 acts as a master regulator of multipotent cardiovascular progenitor specification. Cell Stem Cell 3, 69–84 (2008).

  7. 7.

    et al. Defining the earliest step of cardiovascular progenitor specification during embryonic stem cell differentiation. J. Cell Biol. 192, 751–765 (2011).

  8. 8.

    et al. Mesp1 coordinately regulates cardiovascular fate restriction and epithelial-mesenchymal transition in differentiating ESCs. Cell Stem Cell 3, 55–68 (2008).

  9. 9.

    et al. MesP1 drives vertebrate cardiovascular differentiation through Dkk-1-mediated blockade of Wnt-signalling. Nat. Cell Biol. 10, 338–345 (2008).

  10. 10.

    & Tracing cells for tracking cell lineage and clonal behavior. Dev. Cell 21, 394–409 (2011).

  11. 11.

    & The fate diversity of mesodermal cells within the heart field during chicken early embryogenesis. Dev. Biol. 177, 265–273 (1996).

  12. 12.

    & Fate diversity of primitive streak cells during heart field formation in ovo. Dev. Dyn. 219, 505–513 (2000).

  13. 13.

    et al. Developmental origin of a bipotential myocardial and smooth muscle cell precursor in the mammalian heart. Cell 127, 1137–1150 (2006).

  14. 14.

    et al. Multipotent embryonic isl1+ progenitor cells lead to cardiac, smooth muscle, and endothelial cell diversification. Cell 127, 1151–1165 (2006).

  15. 15.

    , & Multipotent flk-1+ cardiovascular progenitor cells give rise to the cardiomyocyte, endothelial, and vascular smooth muscle lineages. Dev. Cell 11, 723–732 (2006).

  16. 16.

    et al. Transcriptional regulation of Mesp1 and Mesp2 genes: differential usage of enhancers during development. Mech. Dev. 108, 59–69 (2001).

  17. 17.

    et al. MesP1: a novel basic helix-loop-helix protein expressed in the nascent mesodermal cells during mouse gastrulation. Development 122, 2769–2778 (1996).

  18. 18.

    , , , & Mesp1-nonexpressing cells contribute to the ventricular cardiac conduction system. Dev. Dyn. 235, 395–402 (2006).

  19. 19.

    , & Building the mammalian heart from two sources of myocardial cells. Nat. Rev. Genet. 6, 826–835 (2005).

  20. 20.

    et al. Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143, 134–144 (2010).

  21. 21.

    , , , & Lineage tree for the venous pole of the heart: clonal analysis clarifies controversial genealogy based on genetic tracing. Circ. Res. 111, 1313–1322 (2012).

  22. 22.

    et al. Distinct origins and genetic programs of head muscle satellite cells. Dev. Cell 16, 822–832 (2009).

  23. 23.

    et al. Distinct regulatory cascades govern extraocular and pharyngeal arch muscle progenitor cell fates. Dev. Cell 16, 810–821 (2009).

  24. 24.

    et al. Clonal analysis reveals common lineage relationships between head muscles and second heart field derivatives in the mouse embryo. Development 137, 3269–3279 (2010).

  25. 25.

    & Development of the endocardium. Pediatr. Cardiol. 31, 391–399 (2010).

  26. 26.

    et al. Isl1 identifies a cardiac progenitor population that proliferates prior to differentiation and contributes a majority of cells to the heart. Dev. Cell 5, 877–889 (2003).

  27. 27.

    et al. Efficient Cre-mediated deletion in cardiac progenitor cells conferred by a 3’UTR-ires-Cre allele of the homeobox gene Nkx2-5. Int. J. Dev. Biol. 46, 431–439 (2002).

  28. 28.

    et al. HCN4 dynamically marks the first heart field and conduction system precursors. Circ. Res. 113, 399–407 (2013).

  29. 29.

    et al. A HCN4+ cardiomyogenic progenitor derived from the first heart field and human pluripotent stem cells. Nat. Cell Biol.1098–1106 (2013).

  30. 30.

    et al. The heart endocardium is derived from vascular endothelial progenitors. Development 138, 4777–4787 (2011).

  31. 31.

    & Vascularizing the heart. Cardiovasc. Res. 91, 260–268 (2011).

  32. 32.

    , & Reassessment of Isl1 and Nkx2-5 cardiac fate maps using a Gata4-based reporter of Cre activity. Dev. Biol. 323, 98–104 (2008).

  33. 33.

    et al. Epicardial progenitors contribute to the cardiomyocyte lineage in the developing heart. Nature 454, 109–113 (2008).

  34. 34.

    , , , & Nkx2-5- and Isl1-expressing cardiac progenitors contribute to proepicardium. Biochem. Biophys. Res. Commun. 375, 450–453 (2008).

  35. 35.

    et al. Eomesodermin induces Mesp1 expression and cardiac differentiation from embryonic stem cells in the absence of Activin. EMBO Rep. 13, 355–362 (2012).

  36. 36.

    et al. The T-box transcription factor Eomesodermin acts upstream of Mesp1 to specify cardiac mesoderm during mouse gastrulation. Nat. Cell Biol. 13, 1084–1091 (2011).

  37. 37.

    et al. The transcription/migration interface in heart precursors of Ciona intestinalis. Science 320, 1349–1352 (2008).

  38. 38.

    , , & Mutation of l7Rn3 shows that Odz4 is required for mouse gastrulation. Genetics 169, 285–299 (2005).

  39. 39.

    , & Mouse Tenm4 is required for mesoderm induction. BMC Dev. Biol. 13, 9 (2013).

  40. 40.

    et al. Induction and patterning of trunk and tail neural ectoderm by the homeobox gene eve1 in zebrafish embryos. Proc. Natl Acad. Sci. USA 107, 3564–3569 (2010).

  41. 41.

    et al. Mixl1 is required for axial mesendoderm morphogenesis and patterning in the murine embryo. Development 129, 3597–3608 (2002).

  42. 42.

    et al. OTX1 compensates for OTX2 requirement in regionalisation of anterior neuroectoderm. Gene. Expr. Patterns 3, 497–501 (2003).

  43. 43.

    & Analysis of mouse Evx genes: Evx-1 displays graded expression in the primitive streak. Dev. Biol. 151, 273–287 (1992).

  44. 44.

    et al. Lim1 activity is required for intermediate mesoderm differentiation in the mouse embryo. Dev. Biol. 223, 77–90 (2000).

  45. 45.

    et al. Embryonic retinoic acid synthesis is essential for heart morphogenesis in the mouse. Development 128, 1019–1031 (2001).

  46. 46.

    et al. RXR alpha mutant mice establish a genetic basis for vitamin A signaling in heart morphogenesis. Genes Dev. 8, 1007–1018 (1994).

  47. 47.

    et al. Foxh1 is essential for development of the anterior heart field. Dev. Cell 7, 331–345 (2004).

  48. 48.

    et al. Hox genes define distinct progenitor sub-domains within the second heart field. Dev. Biol. 353, 266–274 (2011).

  49. 49.

    et al. Baf60c is essential for function of BAF chromatin remodelling complexes in heart development. Nature 432, 107–112 (2004).

  50. 50.

    , , & The murine winged helix transcription factors, Foxc1 and Foxc2, are both required for cardiovascular development and somitogenesis. Genes Dev. 15, 2470–2482 (2001).

  51. 51.

    , & Msg1 and Mrg1, founding members of a gene family, show distinct patterns of gene expression during mouse embryogenesis. Mech. Dev. 72, 27–40 (1998).

  52. 52.

    et al. Hypomorphic Mesp allele distinguishes establishment of rostrocaudal polarity and segment border formation in somitogenesis. Development 129, 2473–2481 (2002).

  53. 53.

    & Forkhead transcription factors, Foxc1 and Foxc2, are required for the morphogenesis of the cardiac outflow tract. Dev. Biol. 296, 421–436 (2006).

  54. 54.

    , , , & Chamber specification of atrial myosin light chain-2 expression precedes septation during murine cardiogenesis. J. Biol. Chem. 269, 16961–16970 (1994).

  55. 55.

    et al. Nkx2-5 transactivates the Ets-related protein 71 gene and specifies an endothelial/endocardial fate in the developing embryo. Proc. Natl Acad. Sci. USA 106, 814–819 (2009).

  56. 56.

    et al. Etv2/ER71 induces vascular mesoderm from Flk1+PDGFRα+ primitive mesoderm. Blood 118, 6975–6986 (2011).

  57. 57.

    et al. ER71 directs mesodermal fate decisions during embryogenesis. Development 138, 4801–4812 (2011).

  58. 58.

    et al. Vascular endothelial and endocardial progenitors differentiate as cardiomyocytes in the absence of Etsrp/Etv2 function. Development 138, 4721–4732 (2011).

  59. 59.

    et al. Endocardial cells are a distinct endothelial lineage derived from Flk1+ multipotent cardiovascular progenitors. Dev. Biol. 333, 78–89 (2009).

  60. 60.

    , , , & Early restriction of peripheral and proximal cell lineages during formation of the lung. Proc. Natl Acad. Sci. USA 99, 10482–10487 (2002).

  61. 61.

    et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 13, 133–140 (2010).

  62. 62.

    et al. Defining the epithelial stem cell niche in skin. Science 303, 359–363 (2004).

  63. 63.

    , , , & An optimized procedure for whole-mount in situ hybridization on mouse embryos and embryoid bodies. Nat. Protoc. 3, 1194–1201 (2008).

  64. 64.

    , , , & Fate mapping of the mouse midbrain-hindbrain constriction using a site-specific recombination system. Curr. Biol. 8, 665–668 (1998).

  65. 65.

    et al. Accurate expression profiling of very small cell populations. PLoS ONE 5, e14418 (2010).

  66. 66.

    , , & Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol. 10, R25 (2009).

  67. 67.

    et al. Model-based analysis of ChIP-Seq (MACS). Genome Biol. 9, R137 (2008).

  68. 68.

    et al. Quartz-Seq: a highly reproducible and sensitive single-cell RNA sequencing method, reveals non-genetic gene-expression heterogeneity. Genome Biol. 14, R31 (2013).

  69. 69.

    & Single-cell expression profiling of human epidermal stem and transit-amplifying cells: Lrig1 is a regulator of stem cell quiescence. Proc. Natl Acad. Sci. USA 103, 11958–11963 (2006).

  70. 70.

    et al. Single-cell gene expression profiling reveals functional heterogeneity of undifferentiated human epidermal cells. Development 140, 1433–1444 (2013).

Download references

Acknowledgements

We thank F. Bollet-Quivogne and J-M. Vanderwinden for their help with confocal imaging and M. Tarabichi for his help with gene set enrichment analysis. We thank M. Buckingham and A. Joyner for kindly providing the probes for in situ hybridization. We also thank the Genomic core facility of the EMBL, Heidelberg for their help with the ChIP-Seq. F.L. has been sequentially supported by the FNRS and the EMBO long-term fellowship. S.C. is supported by a fellowship of the FRS/FRIA. X.L. is supported by the FNRS. A.B. is supported by the FRS/FNRS. S.R. and B.D.S. are supported by the Wellcome Trust (grant number 098357/Z/12/Z). C.B. is an investigator of WELBIO. This work was supported by the FNRS, the ULB foundation, the Fondation contre le Cancer, the European Research Council (ERC), and the foundation Bettencourt Schueller (C.B. and F.L.).

Author information

Author notes

    • Fabienne Lescroart
    •  & Samira Chabab

    These authors contributed equally to this work.

Affiliations

  1. Université Libre de Bruxelles, IRIBHM, Brussels B-1070, Belgium

    • Fabienne Lescroart
    • , Samira Chabab
    • , Xionghui Lin
    • , Catherine Paulissen
    • , Christine Dubois
    • , Antoine Bondue
    •  & Cédric Blanpain
  2. Cavendish Laboratory, Department of Physics, J. J. Thomson Avenue, Cambridge CB3 0HE, UK

    • Steffen Rulands
    •  & Benjamin D. Simons
  3. The Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK

    • Steffen Rulands
    •  & Benjamin D. Simons
  4. Functional Genomics Core, Institute for Research in Biomedicine, Barcelona 08028, Spain

    • Annie Rodolosse
    •  & Herbert Auer
  5. Université Catholique de Louvain, de Duve Institute, Brussels B-1200, Belgium

    • Younes Achouri
  6. Department of Cardiology, Hopital Erasme, Brussels B-1070, Belgium

    • Antoine Bondue
  7. WELBIO, Université Libre de Bruxelles, Brussels B-1070, Belgium

    • Cédric Blanpain

Authors

  1. Search for Fabienne Lescroart in:

  2. Search for Samira Chabab in:

  3. Search for Xionghui Lin in:

  4. Search for Steffen Rulands in:

  5. Search for Catherine Paulissen in:

  6. Search for Annie Rodolosse in:

  7. Search for Herbert Auer in:

  8. Search for Younes Achouri in:

  9. Search for Christine Dubois in:

  10. Search for Antoine Bondue in:

  11. Search for Benjamin D. Simons in:

  12. Search for Cédric Blanpain in:

Contributions

C.B., F.L., S.C. and X.L. designed the experiments and performed data analysis. F.L. and S.C. performed most of the experiments. X.L. performed the single-cell PCR analysis. Y.A. generated the Mesp1–rtTA transgenic mice. A.R. and H. A. performed microarrays. C.P. provided technical assistance. C.D. helped with FACS isolation of Mesp1-expressing cells. A.B. helped in the design and initial characterization of the Mesp1–rtTA transgene. B.D.S. and S.R. performed the bio-statistical analysis of the clonal fate data. C.B. and F.L. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Cédric Blanpain.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

Excel files

  1. 1.

    Supplementary table 1

    Supplementary Information

  2. 2.

    Supplementary table 2

    Supplementary Information

  3. 3.

    Supplementary table 3

    Supplementary Information

  4. 4.

    Supplementary table 4

    Supplementary Information

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ncb3024