Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Review Article
  • Published:

Plasticity and reprogramming of differentiated cells in amphibian regeneration

Key Points

  • Regenerative ability is widespread in metazoan phylogeny, but has been lost in many species for reasons that are not understood.

  • The champions of regeneration among adult verterbrates are the urodele amphibians, such as the newt, which can regenerate their limbs, jaws, lens and large sections of the heart.

  • Urodele regeneration depends on the local plasticity of differentiated cells that remain after injury or tissue removal. This involves re-entry to the cell cycle and loss of differentiated characteristics, so as to generate a local progenitor cell of restricted potentiality.

  • The molecular cell biology of urodele plasticity has been investigated in detail in the skeletal myofibre and myotube. Newt myotubes can re-enter the cell cycle by activating a pathway that leads to phosphorylation of the retinoblastoma protein pRb. This pathway is triggered by a ligand that is generated downstream of thrombin activation to which mammalian myotubes are completely unresponsive.

  • Newt myotubes are converted to mononucleate progeny after implantation into the regenerating limb. Mouse myotubes are also converted to mononucleate cells by expression of the Msx-1 gene, by exposure to a substituted purine known as myoseverin or to extracts of a regenerating newt limb.

  • Our understanding of re-programming during urodele regeneration might lead to a marked enhancement of regenerative ability in mammals.

Abstract

Adult urodele amphibians, such as the newt, can regenerate their limbs and various other structures. This is the result of the plasticity and reprogramming of residual differentiated cells, rather than the existence of a 'reserve-cell' mechanism. The recent demonstrations of plasticity in mouse myotubes should facilitate comparative studies of the pathways that underlie the regenerative response, as well as proposing new approaches to promote mammalian regeneration.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Purchase on Springer Link

Instant access to full article PDF

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Urodele limb regeneration.
Figure 2: Plasticity of differentiated cells during regeneration of newt heart or lens.
Figure 3: Plasticity of urodele myogenesis.
Figure 4: Thrombin activity counteracts post-mitotic arrest in newt myotubes.
Figure 5: Arrested myotubes undergo cellularization after implantation into the newt blastema.
Figure 6: Categories of genes that are regulated after myoseverin treatment of myotubes or serum stimulation of fibroblasts.

Similar content being viewed by others

Zixuan Zhao, Xinyi Chen, … Hanry Yu

References

  1. Dinsmore, C. E. A History of Regeneration Research (Cambridge Univ. Press, Cambridge, 1991).

    Google Scholar 

  2. Brockes, J. P. Amphibian limb regeneration: rebuilding a complex structure. Science 276, 81–87 (1997).An overview of urodele regeneration that focuses on plasticity and positional identity.

    Article  CAS  PubMed  Google Scholar 

  3. Brockes, J. P., Kumar, A. & Velloso, C. P. Regeneration as an evolutionary variable. J. Anat. 199, 3–11 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Pecorino, L. T., Entwistle, A. & Brockes, J. P. Activation of a single retinoic acid receptor isoform mediates proximodistal respecification. Curr. Biol. 6, 563–569 (1996).

    Article  CAS  PubMed  Google Scholar 

  5. Torok, M. A., Gardiner, D. M., Shubin, N. H. & Bryant, S. V. Expression of HoxD genes in developing and regenerating axolotl limbs. Dev. Biol. 200, 225–233 (1998).

    Article  CAS  PubMed  Google Scholar 

  6. Maden, M. Vitamin A and pattern formation in the regenerating limb. Nature 295, 672–675 (1982).

    Article  CAS  PubMed  Google Scholar 

  7. Nardi, J. B. & Stocum, D. L. Surface properties of regenerating limb cells: evidence for gradation along the proximodistal axis. Differentiation 25, 27–31 (1983).

    Article  Google Scholar 

  8. Raff, R. A. The Shape of Life (Univ. Chicago, Chicago, 1996).

    Book  Google Scholar 

  9. Carroll, S. B. Chance and necessity: the evolution of morphological complexity and diversity. Nature 409, 1102–1109 (2001).

    Article  CAS  PubMed  Google Scholar 

  10. Alvarado, A. S. Regeneration in the metazoans: why does it happen? Bioessays 22, 578–590 (2000).This reviews the evolutionary questions about regeneration and its origins, such as why some animals regenerate but others apparently do not.

    Article  CAS  Google Scholar 

  11. Goss, R. J. Principles of Regeneration (Academic, New York, 1969).

    Google Scholar 

  12. Ghosh, S., Thorogood, P. & Ferretti, P. Regenerative capability of upper and lower jaws in the newt. Int. J. Dev. Biol. 38, 479–490 (1994).

    CAS  PubMed  Google Scholar 

  13. Reyer, R. W. Regeneration of the lens in the amphibian eye. Quart. Rev. Biol. 29, 1–46 (1954).

    Article  CAS  PubMed  Google Scholar 

  14. Mitashov, V. I. Mechanisms of retina regeneration in urodeles. Int. J. Dev. Biol. 40, 833–844 (1996).

    CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  16. Oberpriller, J. O., Oberpriller, J. C., Matz, D. G. & Soonpaa, M. H. Stimulation of proliferative events in the adult amphibian cardiac myocyte. Ann. NY Acad. Sci. 752, 30–46 (1995).

    Article  CAS  PubMed  Google Scholar 

  17. Soonpaa, M. H. & Field, L. J. Survey of studies examining mammalian cardiomyocyte DNA synthesis. Circ. Res. 83, 15–26 (1998).

    Article  CAS  PubMed  Google Scholar 

  18. Okada, T. S. Transdifferentiation (Clarendon, Oxford, 1991).

    Google Scholar 

  19. Eguchi, G. Cellular and molecular background of wolffian lens regeneration. Cell Differ. Dev. 25, S147–S158 (1988).

    Article  Google Scholar 

  20. Eguchi, G., Abe, S. I. & Watanabe, K. Differentiation of lens-like structures from newt iris epithelial cells in vitro. Proc. Natl Acad. Sci. USA 71, 5052–5056 (1974).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Eguchi, G. & Okada, T. S. Differentiation of lens tissue from the progeny of chick retinal pigment cells cultured in vitro: a demonstration of a switch of cell types in clonal cell culture. Proc. Natl Acad. Sci. USA 70, 1495–1499 (1973).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Tsonis, P. A. Limb Regeneration (Cambridge Univ. Press, Cambridge, 1996).

    Google Scholar 

  23. Wallace, H. Vertebrate Limb Regeneration (Wiley and Sons, New York, 1981).

    Google Scholar 

  24. Tanaka, E. M., Gann, A. A., Gates, P. B. & Brockes, J. P. Newt myotubes re-enter the cell cycle by phosphorylation of the retinoblastoma protein. J. Cell Biol. 136, 155–165 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Tanaka, E. M., Drechsel, D. N. & Brockes, J. P. Thrombin regulates S-phase re-entry by cultured newt myotubes. Curr. Biol. 9, 792–799 (1999).References 24 and 25 provide the first clear evidence that a urodele differentiated cell — the skeletal myotube — is intrinsically different from its mammalian counterpart.

    Article  CAS  PubMed  Google Scholar 

  26. Velloso, C. P., Simon, A. & Brockes, J. P. Mammalian postmitotic nuclei re-enter the cell cycle after serum stimulation in newt/mouse hybrid myotubes. Curr. Biol. 11, 855–858 (2001).

    Article  CAS  PubMed  Google Scholar 

  27. Kumar, A., Velloso, C. P., Imokawa, Y. & Brockes, J. P. Plasticity of retrovirus-labelled myotubes in the newt limb regeneration blastema. Dev. Biol. 218, 125–136 (2000).

    Article  CAS  PubMed  Google Scholar 

  28. Lo, D. C., Allen, F. & Brockes, J. P. Reversal of muscle differentiation during urodele limb regeneration. Proc. Natl Acad. Sci. USA 90, 7230–7234 (1993).This study found that implanted labelled myotubes underwent reversal of muscle differentiation in the limb blastema.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Velloso, C. P., Kumar, A., Tanaka, E. M. & Brockes, J. P. Generation of mononucleate cells from post-mitotic myotubes proceeds in the absence of cell cycle progression. Differentiation 66, 239–246 (2000).

    Article  CAS  PubMed  Google Scholar 

  30. Echeverri, K., Clarke, J. D. & Tanaka, E. M. In vivo imaging indicates muscle fiber dedifferentiation is a major contributor to the regenerating tail blastema. Dev. Biol. 236, 151–164 (2001).

    Article  CAS  PubMed  Google Scholar 

  31. Steen, T. P. Stability of chondrocyte differentiation and contribution of muscle to cartilage during limb regeneration in the axolotl (Siredon mexicanum). J. Exp. Zool. 167, 49–78 (1968).

    Article  CAS  PubMed  Google Scholar 

  32. Reyer, R. W., Woolfitt, R. A. & Withersty, L. T. Stimulation of lens regeneration from the newt dorsal iris when implanted into the blastema of the regenerating limb. Dev. Biol. 32, 258–281 (1973).

    Article  CAS  PubMed  Google Scholar 

  33. Ito, M., Hayashi, T., Kuroiwa, A. & Okamoto, M. Lens formation by pigmented epithelial cell reaggregate from dorsal iris implanted into limb blastema in the adult newt. Dev. Growth Differ. 41, 429–440 (1999).

    Article  CAS  PubMed  Google Scholar 

  34. Kim, W. S. & Stocum, D. L. Retinoic acid modifies positional memory in the anteroposterior axis of regenerating axolotl limbs. Dev. Biol. 114, 170–179 (1986).

    Article  CAS  PubMed  Google Scholar 

  35. Clarke, D. L. et al. Generalized potential of adult neural stem cells. Science 288, 1660–1663 (2000).

    Article  CAS  PubMed  Google Scholar 

  36. Michalopoulos, G. K. & DeFrances, M. C. Liver regeneration. Science 276, 60–66 (1997).

    Article  CAS  PubMed  Google Scholar 

  37. Aguayo, A. J., Epps, J., Charron, L. & Bray, G. M. Multipotentiality of Schwann cells in cross-anastomosed and grafted myelinated and unmyelinated nerves: quantitative microscopy and radioautography. Brain Res. 104, 1–20 (1976).

    Article  CAS  PubMed  Google Scholar 

  38. Weinberg, H. J. & Spencer, P. S. Studies on the control of myelinogenesis. II. Evidence for neuronal regulation of myelin production. Brain Res. 113, 363–378 (1976).

    Article  CAS  PubMed  Google Scholar 

  39. Olwin, B. B. & Hauschka, S. D. Cell surface fibroblast growth factor and epidermal growth factor receptors are permanently lost during skeletal muscle terminal differentiation in culture. J. Cell Biol. 107, 761–769 (1988).

    Article  CAS  PubMed  Google Scholar 

  40. Ferretti, P. & Brockes, J. P. Culture of newt cells from different tissues and their expression of a regeneration-associated antigen. J. Exp. Zool. 247, 77–91 (1988).

    Article  CAS  PubMed  Google Scholar 

  41. Schneider, J. W., Gu, W., Zhu, L., Mahdavi, V. & Nadal-Ginard, B. Reversal of terminal differentiation mediated by p107 in Rb−/− muscle cells. Science 264, 1467–1471 (1994).

    Article  CAS  PubMed  Google Scholar 

  42. Novitch, B. G., Mulligan, G. J., Jacks, T. & Lassar, A. B. Skeletal muscle cells lacking the retinoblastoma protein display defects in muscle gene expression and accumulate in S and G2 phases of the cell cycle. J. Cell Biol. 135, 441–456 (1996).

    Article  CAS  PubMed  Google Scholar 

  43. Zacksenhaus, E. et al. pRb controls proliferation, differentiation, and death of skeletal muscle cells and other lineages during embryogenesis. Genes Dev. 10, 3051–3064 (1996).

    Article  CAS  PubMed  Google Scholar 

  44. Novitch, B. G., Spicer, D. B., Kim, P. S., Cheung, W. L. & Lassar, A. B. pRb is required for MEF2-dependent gene expression as well as cell-cycle arrest during skeletal muscle differentiation. Curr. Biol. 9, 449–459 (1999).

    Article  CAS  PubMed  Google Scholar 

  45. Solari, F. et al. Multinucleated cells can continuously generate mononucleated cells in the absence of mitosis: a study of cells of the avian osteoclast lineage. J. Cell Sci. 108, 3233–3241 (1995).

    CAS  PubMed  Google Scholar 

  46. Odelberg, S. J., Kollhoff, A. & Keating, M. T. Dedifferentiation of mammalian myotubes induced by msx1. Cell 103, 1099–1109 (2000).This study showed that the expression of Msx-1 in mouse myotubes leads to the generation of mononucleate pluripotent progeny.

    Article  CAS  PubMed  Google Scholar 

  47. Hu, G., Lee, H., Price, S. M., Shen, M. M. & Abate-Shen, C. Msx homeobox genes inhibit differentiation through upregulation of cyclin D1. Development 128, 2373–2384 (2001).

    CAS  PubMed  Google Scholar 

  48. Song, K., Wang, Y. & Sassoon, D. Expression of Hox-7.1 in myoblasts inhibits terminal differentiation and induces cell transformation. Nature 360, 477–481 (1992).

    Article  CAS  PubMed  Google Scholar 

  49. Woloshin, P. et al. MSX1 inhibits myoD expression in fibroblast x 10T1/2 cell hybrids. Cell 82, 611–620 (1995).

    Article  CAS  PubMed  Google Scholar 

  50. Koshiba, K., Kuroiwa, A., Yamamoto, H., Tamura, K. & Ide, H. Expression of Msx genes in regenerating and developing limbs of axolotl. J. Exp. Zool. 282, 703–714 (1998).

    Article  CAS  PubMed  Google Scholar 

  51. Rosania, G. R. et al. Myoseverin, a microtubule-binding molecule with novel cellular effects. Nature Biotechnol. 18, 304–308 (2000).This study isolated a trisubstituted purine from a combinatorial library that induces cellularization of mouse myotubes.

    Article  CAS  Google Scholar 

  52. Perez, O. D., Chang, Y. T., Rosania, G., Sutherlin, D. & Schultz, P. G. Inhibition and reversal of myogenic differentiation by purine-based microtubule assembly inhibitors. Chem. Biol. 9, 475–483 (2002).

    Article  CAS  PubMed  Google Scholar 

  53. Iyer, V. R. et al. The transcriptional program in the response of human fibroblasts to serum. Science 283, 83–87 (1999).

    Article  CAS  PubMed  Google Scholar 

  54. McGann, C. J., Odelberg, S. J. & Keating, M. T. Mammalian myotube dedifferentiation induced by newt regeneration extract. Proc. Natl Acad. Sci. USA 98, 13699–13704 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Agata, K. et al. Genetic characterization of the multipotent dedifferentiated state of pigmented epithelial cells in vitro. Development 118, 1025–1030 (1993).

    CAS  PubMed  Google Scholar 

  56. Heber-Katz, E. The regenerating mouse ear. Semin. Cell Dev. Biol. 10, 415–419 (1999).

    Article  CAS  PubMed  Google Scholar 

  57. Clark, L. D., Clark, R. K. & Heber-Katz, E. A new murine model for mammalian wound repair and regeneration. Clin. Immunol. Immunopathol. 88, 35–45 (1998).

    Article  CAS  PubMed  Google Scholar 

  58. Leferovich, J. M. et al. Heart regeneration in adult MRL mice. Proc. Natl Acad. Sci. USA 98, 9830–9835 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. McBrearty, B. A., Clark, L. D., Zhang, X. M., Blankenhorn, E. P. & Heber-Katz, E. Genetic analysis of a mammalian wound-healing trait. Proc. Natl Acad. Sci. USA 95, 11792–11797 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Bianco, P. & Robey, P. G. Stem cells in tissue engineering. Nature 414, 118–121 (2001).

    Article  CAS  PubMed  Google Scholar 

  61. Hughes, R. N. A Functional Biology of Clonal Animals (Chapman and Hall, London, 1989).

    Google Scholar 

  62. Newth, D. R. New (and Better?) Parts for Old (eds Johnson, M. L., Abercrombie, M. & Fogg, G. E.) (Penguin, Middlesex, 1958).

    Google Scholar 

  63. Johnson, S. L. & Weston, J. A. Temperature-sensitive mutations that cause stage-specific defects in Zebrafish fin regeneration. Genetics 141, 1583–1595 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  64. Tosh, D. & Slack, J. M. How cells change their phenotype. Nature Rev. Mol. Cell Biol. 3, 187–194 (2002).

    Article  CAS  Google Scholar 

  65. Tsai, R. Y., Kittappa, R. & McKay, R. D. Plasticity, niches, and the use of stem cells. Dev. Cell 2, 707–712 (2002).

    Article  CAS  PubMed  Google Scholar 

  66. Iten, L. & Bryant, S. V. Forelimb regeneration from different levels of amputation in the newt N. viridescens. Length, rate and stages. Wilhelm Roux Arch. Dev. Biol. 173, 263–282 (1973).

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jeremy P. Brockes.

Related links

Related links

DATABASES

LocusLink

CDK4

EGF

Msx-1

pRb

prothrombin

Swiss-Prot

GFP

p16INK4

FURTHER READING

Encyclopedia of Life Sciences

Regeneration: growth factors in limb development

Regeneration of the urodele limb

Regeneration of the vertebrate lens and other eye structures

Regeneration of the vertebrate tail

Regeneration of vertebrate appendages

Regeneration of vertebrate tissues: model systems

Regeneration: principles

Spallanzani

Glossary

URODELE

An order of the class Amphibia, which comprises newts and salamanders, which have elongated bodies, short limbs and a tail.

METAZOAN

Refers to the kingdom Animalia (animals), which comprises 35 phyla of multicellular organisms.

PHYLOGENY

Evolutionary history that is sometimes represented by the hypothesized ancestor–descendant relationship of a group of organisms.

CARDIOMYOCYTE

A muscle cell of the heart.

S (SYNTHESIS) PHASE

The phase of the eukaryotic cell cycle in which DNA is synthesized.

MYOFIBRIL

The structural unit of striated muscle fibres. Several myofibrils make up each fibre.

MYOTUBE

The multinucleate structure that is formed by the fusion of proliferating myoblasts and is characterized by the presence of certain muscle-specific marker proteins.

MYOFIBRE

A skeletal-muscle fibre that consists of one long multinucleate cell.

PLURIPOTENT CELL

A stem cell that can give rise to more than one differentiated cell type.

HEPATOCYTES

The parenchymal cells of the liver that are responsible for the synthesis, degradation and storage of a wide range of substances.

ROTIFERA

A small phylum of microscopic multicellular organisms. They have a wheel-like ciliated organ (from which they derive their name) that they use for swimming and feeding.

TURBELLARIANS

A class of platyhelminthes that comprises mostly aquatic and free-living organisms. They have a ciliated epidermis for locomotion and a simple gut.

ANNELID

A segmented worm.

PLANARIAN

Describes free-living members of the invertebrate phylum platyhelminthes.

SCHWANN CELL

A cell that produces myelin and ensheathes axons in the peripheral nervous system.

MYELIN

Proteins that are produced by Schwann cells or oligodendrocytes that cause adjacent plasma membranes to stack tightly together.

MESENCHYME

Immature connective tissue that consists of cells that are embedded in extracellular matrix.

GREEN FLUORESCENT PROTEIN

An autofluorescent protein that was originally isolated from the jellyfish Aequorea victoria. It can be genetically conjugated with proteins to make them fluorescent. The most widely used mutant, EGFP, has an emission maximum at 510 nm.

5′ BROMODEOXYURIDINE

(BrdU). A base analogue of thymidine, which is often used experimentally to label dividing cells.

NOMARSKI OPTICS

Also known as differential interference contrast microscopy, this technique forms images of high contrast and resolution in unstained cells using birefringent prisms and polarized light.

EXOCRINE CELL

A cell that makes up part of an exocrine gland, which discharges its secretion through a duct.

G2

The phase of the cell cycle through which cells progress after S phase but before M phase.

HETEROKARYON

A cell that contains two nuclei in a common cytoplasm.

CYTOKINESIS

The process of cytoplasmic division.

OSTEOCLAST

A mesenchymal cell that can differentiate into a bone-degrading cell.

GIANT CELLS

Large multinucleated cells that are thought to result from the fusion of macrophages.

CHONDROGENIC

Able to form cartilage.

ADIPOGENIC

Able to form fat or adipose tissue.

MICROTUBULE

A hollow tube, 25 nm in diameter, that is formed by the lateral association of 13 protofilaments, which are themselves polymers of α- and β-tubulin subunits.

DNA MICROARRAYS

Devices that are used to analyse complex nucleic acid samples by hybridization. They make it possible to quantitate the amount of different nucleic acid molecules that are present in a sample of interest.

CRYOGENIC INFARCTION

Obstruction of the blood supply as a result of extremely low temperatures.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Brockes, J., Kumar, A. Plasticity and reprogramming of differentiated cells in amphibian regeneration. Nat Rev Mol Cell Biol 3, 566–574 (2002). https://doi.org/10.1038/nrm881

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nrm881

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing