Letter | Published:

Coordinating cardiomyocyte interactions to direct ventricular chamber morphogenesis

Nature volume 534, pages 700704 (30 June 2016) | Download Citation

Abstract

Many organs are composed of complex tissue walls that are structurally organized to optimize organ function. In particular, the ventricular myocardial wall of the heart comprises an outer compact layer that concentrically encircles the ridge-like inner trabecular layer. Although disruption in the morphogenesis of this myocardial wall can lead to various forms of congenital heart disease1 and non-compaction cardiomyopathies2, it remains unclear how embryonic cardiomyocytes assemble to form ventricular wall layers of appropriate spatial dimensions and myocardial mass. Here we use advanced genetic and imaging tools in zebrafish to reveal an interplay between myocardial Notch and Erbb2 signalling that directs the spatial allocation of myocardial cells to their proper morphological positions in the ventricular wall. Although previous studies have shown that endocardial Notch signalling non-cell-autonomously promotes myocardial trabeculation through Erbb2 and bone morphogenetic protein (BMP) signalling3, we discover that distinct ventricular cardiomyocyte clusters exhibit myocardial Notch activity that cell-autonomously inhibits Erbb2 signalling and prevents cardiomyocyte sprouting and trabeculation. Myocardial-specific Notch inactivation leads to ventricles of reduced size and increased wall thickness because of excessive trabeculae, whereas widespread myocardial Notch activity results in ventricles of increased size with a single-cell-thick wall but no trabeculae. Notably, this myocardial Notch signalling is activated non-cell-autonomously by neighbouring Erbb2-activated cardiomyocytes that sprout and form nascent trabeculae. Thus, these findings support an interactive cellular feedback process that guides the assembly of cardiomyocytes to morphologically create the ventricular myocardial wall and more broadly provide insight into the cellular dynamics of how diverse cell lineages organize to create form.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , , & Genetics of congenital heart disease: the glass half empty. Circ. Res. 112, 707–720 (2013)

  2. 2.

    et al. The relationship of left ventricular trabeculation to ventricular function and structure over a 9.5-year follow-up: the MESA study. J. Am. Coll. Cardiol. 64, 1971–1980 (2014)

  3. 3.

    et al. Notch signaling is essential for ventricular chamber development. Dev. Cell 12, 415–429 (2007)

  4. 4.

    & Clonally dominant cardiomyocytes direct heart morphogenesis. Nature 484, 479–484 (2012)

  5. 5.

    et al. High-resolution imaging of cardiomyocyte behavior reveals two distinct steps in ventricular trabeculation. Development 141, 585–593 (2014)

  6. 6.

    & Social interactions among epithelial cells during tracheal branching morphogenesis. Nature 441, 746–749 (2006)

  7. 7.

    & Notch signalling limits angiogenic cell behaviour in developing zebrafish arteries. Nature 445, 781–784 (2007)

  8. 8.

    et al. Loss of Llgl1 in retinal neuroepithelia reveals links between apical domain size, Notch activity and neurogenesis. Development 139, 1599–1610 (2012)

  9. 9.

    et al. Cardiac contraction activates endocardial Notch signaling to modulate chamber maturation in zebrafish. Development 142, 4080–4091 (2015)

  10. 10.

    et al. A dual role for ErbB2 signaling in cardiac trabeculation. Development 137, 3867–3875 (2010)

  11. 11.

    , , , & Trabecular myocytes of the embryonic heart require N-cadherin for migratory unit identity. Dev. Biol. 193, 1–9 (1998)

  12. 12.

    et al. Notch-responsive cells initiate the secondary transition in larval zebrafish pancreas. Mech. Dev. 126, 898–912 (2009)

  13. 13.

    et al. Notch signaling regulates cardiomyocyte proliferation during zebrafish heart regeneration. Proc. Natl Acad. Sci. USA 111, 1403–1408 (2014)

  14. 14.

    et al. Hepatocyte growth factor signaling in intrapancreatic ductal cells drives pancreatic morphogenesis. PLoS Genet. 9, e1003650 (2013)

  15. 15.

    , & Different levels of Notch signaling regulate quiescence, renewal and differentiation in pancreatic endocrine progenitors. Development 139, 1557–1567 (2012)

  16. 16.

    et al. Mutation of 3-hydroxy-3-methylglutaryl CoA synthase I reveals requirements for isoprenoid and cholesterol synthesis in oligodendrocyte migration arrest, axon wrapping, and myelin gene expression. J. Neurosci. 34, 3402–3412 (2014)

  17. 17.

    , & Dependence of cardiac trabeculation on neuregulin signaling and blood flow in zebrafish. Dev. Dyn. 240, 446–456 (2011)

  18. 18.

    et al. erbb3 and erbb2 are essential for Schwann cell migration and myelination in zebrafish. Curr. Biol. 15, 513–524 (2005)

  19. 19.

    & Dynamic smad-mediated BMP signaling revealed through transgenic zebrafish. Dev. Dyn. 240, 712–722 (2011)

  20. 20.

    et al. BMP10 is essential for maintaining cardiac growth during murine cardiogenesis. Development 131, 2219–2231 (2004)

  21. 21.

    Lateral inhibition during vulval induction in Caenorhabditis elegans. Nature 335, 551–554 (1988)

  22. 22.

    & The choice of cell fate in the epidermis of Drosophila. Cell 64, 1083–1092 (1991)

  23. 23.

    , , & tal1 regulates the formation of intercellular junctions and the maintenance of identity in the endocardium. Dev. Biol. 383, 214–226 (2013)

  24. 24.

    et al. Notch signaling regulates murine atrioventricular conduction and the formation of accessory pathways. J. Clin. Invest. 121, 525–533 (2011)

  25. 25.

    et al. Numb family proteins are essential for cardiac morphogenesis and progenitor differentiation. Development 141, 281–295 (2014)

  26. 26.

    et al. Inhibition of Notch2 by Numb/Numblike controls myocardial compaction in the heart. Cardiovasc. Res. 96, 276–285 (2012)

  27. 27.

    , , & Hey genes: a novel subfamily of hairy- and Enhancer of split related genes specifically expressed during mouse embryogenesis. Mech. Dev. 85, 173–177 (1999)

  28. 28.

    et al. Alagille syndrome is caused by mutations in human Jagged1, which encodes a ligand for Notch1. Nature Genet. 16, 243–251 (1997)

  29. 29.

    et al. Mutations in the NOTCH pathway regulator MIB1 cause left ventricular noncompaction cardiomyopathy. Nature Med. 19, 193–201 (2013)

  30. 30.

    et al. Foxn4 directly regulates tbx2b expression and atrioventricular canal formation. Genes Dev. 22, 734–739 (2008)

  31. 31.

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

  32. 32.

    , , & A mutation in zebrafish hmgcr1b reveals a role for isoprenoids in vertebrate heart-tube formation. Curr. Biol. 17, 252–259 (2007)

  33. 33.

    et al. Primary contribution to zebrafish heart regeneration by gata4+ cardiomyocytes. Nature 464, 601–605 (2010)

  34. 34.

    et al. A systematic genome-wide analysis of zebrafish protein-coding gene function. Nature 496, 494–497 (2013)

  35. 35.

    , , , & Germ-line transmission of a myocardium-specific GFP transgene reveals critical regulatory elements in the cardiac myosin light chain 2 promoter of zebrafish. Dev. Dyn. 228, 30–40 (2003)

  36. 36.

    et al. I-SceI meganuclease mediates highly efficient transgenesis in fish. Mech. Dev. 118, 91–98 (2002)

  37. 37.

    et al. Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish. Development 138, 169–177 (2011)

  38. 38.

    et al. The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. Dev. Dyn. 236, 3088–3099 (2007)

  39. 39.

    et al. Latent TGF-β binding protein 3 identifies a second heart field in zebrafish. Nature 474, 645–648 (2011)

  40. 40.

    et al. In vivo cardiac reprogramming contributes to zebrafish heart regeneration. Nature 498, 497–501 (2013)

  41. 41.

    et al. Dorsomorphin inhibits BMP signals required for embryogenesis and iron metabolism. Nature Chem. Biol. 4, 33–41 (2008)

Download references

Acknowledgements

We thank N. Tedeschi for fish care; B. Le for experimental assistance; S. Evans, D. Yelon, and Chi laboratory members for comments on the manuscript; N. Ninov and D. Stainier for plasmids; B. Link for the d2GFP BMP and d2GFP Notch reporter lines; N. Lawson for the eGFP Notch reporter line; B. Appel for the myocardial Cerulean line; K. Poss for the myocardial CreER and Brainbow/priZm lines; and W. Talbot for the erbb2 mutant. This work was supported in part by grants from American Heart Association (14POST20380738) to L.Z.; the March of Dimes (1-FY14-327) to R.A.M.; the NIH/NHLBI (5R01HL127067) to C.G.B. and C.E.B.; and the National Institutes of Health to N.C.C.

Author information

Affiliations

  1. Department of Medicine, Division of Cardiology, University of California, San Diego, La Jolla, California 92093, USA

    • Peidong Han
    • , Joshua Bloomekatz
    • , Jie Ren
    • , Ruilin Zhang
    • , Jonathan D. Grinstein
    •  & Neil C. Chi
  2. Cardiovascular Research Center, Division of Cardiology, Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts 02129, USA

    • Long Zhao
    • , C. Geoffrey Burns
    •  & Caroline E. Burns
  3. Center for Diabetes and Metabolic Diseases, Department of Pediatrics and Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA

    • Ryan M. Anderson
  4. Institute of Genomic Medicine, University of California, San Diego, La Jolla, California 92093, USA

    • Neil C. Chi

Authors

  1. Search for Peidong Han in:

  2. Search for Joshua Bloomekatz in:

  3. Search for Jie Ren in:

  4. Search for Ruilin Zhang in:

  5. Search for Jonathan D. Grinstein in:

  6. Search for Long Zhao in:

  7. Search for C. Geoffrey Burns in:

  8. Search for Caroline E. Burns in:

  9. Search for Ryan M. Anderson in:

  10. Search for Neil C. Chi in:

Contributions

P.H. and N.C.C. conceived the project and the design of the experimental strategy. P.H., J.R., J.B., R.Z., and J.D.G. conducted experiments. L.Z. generated the ubi:RSdnM transgenic line. P.H. and N.C.C. generated and characterized the myl7:Cre transgenic line. C.E.B., C.G.B., and R.A.M. provided key reagents. P.H., J.B. and N.C.C. prepared the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Neil C. Chi.

Reviewer Information Nature thanks B. G. Bruneau and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Extended data

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature18310

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.