Letter | Published:

Zebrafish heart regeneration occurs by cardiomyocyte dedifferentiation and proliferation

Nature volume 464, pages 606609 (25 March 2010) | Download Citation


Although mammalian hearts show almost no ability to regenerate, there is a growing initiative to determine whether existing cardiomyocytes or progenitor cells can be coaxed into eliciting a regenerative response. In contrast to mammals, several non-mammalian vertebrate species are able to regenerate their hearts1,2,3, including the zebrafish4,5, which can fully regenerate its heart after amputation of up to 20% of the ventricle. To address directly the source of newly formed cardiomyocytes during zebrafish heart regeneration, we first established a genetic strategy to trace the lineage of cardiomyocytes in the adult fish, on the basis of the Cre/lox system widely used in the mouse6. Here we use this system to show that regenerated heart muscle cells are derived from the proliferation of differentiated cardiomyocytes. Furthermore, we show that proliferating cardiomyocytes undergo limited dedifferentiation characterized by the disassembly of their sarcomeric structure, detachment from one another and the expression of regulators of cell-cycle progression. Specifically, we show that the gene product of polo-like kinase 1 (plk1) is an essential component of cardiomyocyte proliferation during heart regeneration. Our data provide the first direct evidence for the source of proliferating cardiomyocytes during zebrafish heart regeneration and indicate that stem or progenitor cells are not significantly involved in this process.

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  1. 1.

    & Response of the adult newt ventricle to injury. J. Exp. Zool. 187, 249–253 (1974)

  2. 2.

    , & Heart development and regeneration in urodeles. Int. J. Dev. Biol. 40, 719–725 (1996)

  3. 3.

    Cell cycle reentry of ventricular and atrial cardiomyocytes and cells within the epicardium following amputation of the ventricular apex in the axolotl, Amblystoma mexicanum: confocal microscopic immunofluorescent image analysis of bromodeoxyuridine-labeled nuclei. Anat. Embryol. (Berl.) 205, 235–244 (2002)

  4. 4.

    , & Heart regeneration in zebrafish. Science 298, 2188–2190 (2002)

  5. 5.

    et al. Activation of Notch signaling pathway precedes heart regeneration in zebrafish. Proc. Natl Acad. Sci. USA 100 (Suppl. 1). 11889–11895 (2003)

  6. 6.

    , , & Adult pancreatic β-cells are formed by self-duplication rather than stem-cell differentiation. Nature 429, 41–46 (2004)

  7. 7.

    et al. p38 MAP kinase inhibition enables proliferation of adult mammalian cardiomyocytes. Genes Dev. 19, 1175–1187 (2005)

  8. 8.

    , , & Neuregulin1/ErbB4 signaling induces cardiomyocyte proliferation and repair of heart injury. Cell 138, 257–270 (2009)

  9. 9.

    et al. A dynamic epicardial injury response supports progenitor cell activity during zebrafish heart regeneration. Cell 127, 607–619 (2006)

  10. 10.

    , , , & Gene expression analysis of zebrafish heart regeneration. PLoS Biol. 4, e260 (2006)

  11. 11.

    et al. Transcriptomics approach to investigate zebrafish heart regeneration. J. Cardiovasc. Med. (Hagerstown) 10.2459/JCM.0b013e3283375900 (22 February 2010)

  12. 12.

    et al. Electron microscopy characterization of cardiomyocyte apoptosis in ischemic heart disease. Int. J. Cardiol. 114, 118–120 (2007)

  13. 13.

    Ultrastructural Pathology—An Introduction to Interpretation (Iowa State Univ. Press, 1994)

  14. 14.

    et al. Conditional targeted cell ablation in zebrafish: a new tool for regeneration studies. Dev. Dyn. 236, 1025–1035 (2007)

  15. 15.

    , , , & Re-programming of newt cardiomyocytes is induced by tissue regeneration. J. Cell Sci. 119, 4719–4729 (2006)

  16. 16.

    , & Hibernating myocardium. N. Engl. J. Med. 339, 173–181 (1998)

  17. 17.

    , , , & Adult rabbit cardiomyocytes undergo hibernation-like dedifferentiation when co-cultured with cardiac fibroblasts. Cardiovasc. Res. 51, 230–240 (2001)

  18. 18.

    , & Can the cardiomyocyte cell cycle be reprogrammed? J. Mol. Cell. Cardiol. 42, 706–721 (2007)

  19. 19.

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

  20. 20.

    , , , & 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)

  21. 21.

    & Controlled expression of transgenes introduced by in vivo electroporation. Proc. Natl Acad. Sci. USA 104, 1027–1032 (2007)

  22. 22.

    , , & Temporally-controlled site-specific recombination in zebrafish. PLoS ONE 4, e4640 (2009)

  23. 23.

    , , , & FlEx-based transgenic reporter lines for visualization of Cre and Flp activity in live zebrafish. Genesis 47, 484–491 (2009)

  24. 24.

    et al. Fate mapping using Cited1-CreERT2 mice demonstrates that the cap mesenchyme contains self-renewing progenitor cells and gives rise exclusively to nephronic epithelia. Dev. Biol. 313, 234–245 (2008)

  25. 25.

    Practical Methods in Electron Microscopy (Elsevier North-Holland, 1974)

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We thank M. C. Fabregat, C. Rodriguez Esteban and I. Dubova for technical assistance, and A. Faucherre for constructive criticism of the manuscript. E.S. was the recipient of a pre-doctoral fellowship from the Ministry of Innovation, Universities and Enterprise (DIUE), Generalitat de Catalunya. This work was supported by grants from Fundacion Cellex, the Ipsen Foundation, the G. Harold and Leila Y. Mathers Charitable Foundation, Sanofi-Aventis, the Ministry of Science and Innovation (MICINN), CIBER, and the National Institutes of Health.

Author Contributions C.J., A.R. and J.C.I.B. conceived the project and designed the experiments. C.J. performed the molecular biology and established the transgenic lines. C.J., E.S. and M.R. conducted the experiments. M.M. performed the immunohistochemistry and confocal/transmission electron microscopy imaging. C.J., A.R. and J.C.I.B. wrote the manuscript.

Author information

Author notes

    • Eduard Sleep
    • , Marina Raya
    •  & Angel Raya

    Present addresses: Control of Stem Cell Potency group, Institute for Bioengineering of Catalonia (IBEC), Baldiri Reixac 15, 08028 Barcelona, Spain (E.S., A.R.); Department of Experimental and Health Sciences, Pompeu Fabra University (UPF), Dr Aiguader Street 88, 08003 Barcelona, Spain (M.R.).


  1. Center of Regenerative Medicine in Barcelona,

    • Chris Jopling
    • , Eduard Sleep
    • , Marina Raya
    • , Mercè Martí
    • , Angel Raya
    •  & Juan Carlos Izpisúa Belmonte
  2. Networking Center of Biomedical Research in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Dr Aiguader Street 88, 08003 Barcelona, Spain

    • Eduard Sleep
    • , Angel Raya
    •  & Juan Carlos Izpisúa Belmonte
  3. Institució Catalana de Recerca i Estudis Avançats (ICREA), Passeig Lluis Companys 23, 08010 Barcelona, Spain

    • Angel Raya
  4. Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA

    • Juan Carlos Izpisúa Belmonte


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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Juan Carlos Izpisúa Belmonte.

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