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.

Restoration of anterior regeneration in a planarian with limited regenerative ability

Abstract

Variability of regenerative potential among animals has long perplexed biologists1. On the basis of their exceptional regenerative abilities, planarians have become important models for understanding the molecular basis of regeneration2. However, planarian species with limited regenerative abilities are also found3,4. Despite the importance of understanding the differences between closely related, regenerating and non-regenerating organisms, few studies have focused on the evolutionary loss of regeneration5, and the molecular mechanisms leading to such regenerative loss remain obscure. Here we examine Procotyla fluviatilis, a planarian with restricted ability to replace missing tissues6, using next-generation sequencing to define the gene expression programs active in regeneration-permissive and regeneration-deficient tissues. We found that Wnt signalling is aberrantly activated in regeneration-deficient tissues. Notably, downregulation of canonical Wnt signalling in regeneration-deficient regions restores regenerative abilities: blastemas form and new heads regenerate in tissues that normally never regenerate. This work reveals that manipulating a single signalling pathway can reverse the evolutionary loss of regenerative potential.

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

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Procotyla fluviatilis fails to regenerate heads after amputation in posterior body regions.
Figure 2: Comparative transcriptomics reveal differentially expressed genes following amputation in Reg+ and Reg tissues.
Figure 3: Disruption of Wnt signalling via RNAi rescues regeneration in Reg tissues.

Accession codes

Accessions

BioProject

Gene Expression Omnibus

Data deposits

Sequence read archive (SRA) data reported in this paper were deposited at NCBI as a BioProject under accession number PRJNA205293. RNA-seq analyses have been deposited in the NCBI Gene ExpressionOmnibus under accession number GSE48497.

References

  1. Bely, A. E. Evolutionary loss of animal regeneration: pattern and process. Integr. Comp. Biol. 50, 515–527 (2010)

    Article  Google Scholar 

  2. Newmark, P. A. & Sánchez Alvarado, A. Not your father’s planarian: a classic model enters the era of functional genomics. Nature Rev. Genet. 3, 210–219 (2002)

    CAS  Article  Google Scholar 

  3. Lillie, F. R. Notes on regeneration and regulation in planarians. Am. J. Physiol. 6, 129–141 (1901)

    Article  Google Scholar 

  4. Morgan, T. H. Notes on regeneration. Biol. Bull. 6, 159–172 (1904)

    Article  Google Scholar 

  5. Bely, A. E. & Nyberg, K. G. Evolution of animal regeneration: re-emergence of a field. Trends Ecol. Evol. 25, 161–170 (2010)

    Article  Google Scholar 

  6. Kenk, R. Biota of Freshwater Ecosystems Identification Manual No. 1: Freshwater planarians (Turbellaria) of North America (US Environmental Protection Agency, 1972)

    Google Scholar 

  7. Wenemoser, D., Lapan, S. W., Wilkinson, A. W., Bell, G. W. & Reddien, P. W. A molecular wound response program associated with regeneration initiation in planarians. Genes Dev. 26, 988–1002 (2012)

    CAS  Article  Google Scholar 

  8. Pellettieri, J. et al. Cell death and tissue remodeling in planarian regeneration. Dev. Biol. 338, 76–85 (2010)

    CAS  Article  Google Scholar 

  9. Wenemoser, D. & Reddien, P. W. Planarian regeneration involves distinct stem cell responses to wounds and tissue absence. Dev. Biol. 344, 979–991 (2010)

    CAS  Article  Google Scholar 

  10. Gurley, K. A., Rink, J. C. & Sánchez Alvarado, A. β-catenin defines head versus tail identity during planarian regeneration and homeostasis. Science 319, 323–327 (2008)

    ADS  CAS  Article  Google Scholar 

  11. Petersen, C. P. & Reddien, P. W. Smed-Bcatenin-1 is required for anteroposterior blastema polarity in planarian regeneration. Science 319, 327–330 (2008)

    ADS  CAS  Article  Google Scholar 

  12. Yazawa, S., Umesono, Y., Hayashi, T., Tarui, H. & Agata, K. Planarian Hedgehog/Patched establishes anterior-posterior polarity by regulating Wnt signaling. Proc. Natl Acad. Sci. USA 106, 22329–22334 (2009)

    ADS  CAS  Article  Google Scholar 

  13. Rink, J. C., Gurley, K. A., Elliott, S. A. & Sánchez Alvarado, A. Planarian Hh signaling regulates regeneration polarity and links Hh pathway evolution to cilia. Science 326, 1406–1410 (2009)

    ADS  CAS  Article  Google Scholar 

  14. Gurley, K. A. et al. Expression of secreted Wnt pathway components reveals unexpected complexity of the planarian amputation response. Dev. Biol. 347, 24–39 (2010)

    CAS  Article  Google Scholar 

  15. Reddien, P. W. & Sánchez Alvarado, A. Fundamentals of planarian regeneration. Annu. Rev. Cell Dev. Biol. 20, 725–757 (2004)

    CAS  Article  Google Scholar 

  16. Forsthoefel, D. J., Park, A. E. & Newmark, P. A. Stem cell-based growth, regeneration, and remodeling of the planarian intestine. Dev. Biol. 356, 445–459 (2011)

    CAS  Article  Google Scholar 

  17. Cebrià, F. et al. FGFR-related gene nou-darake restricts brain tissues to the head region of planarians. Nature 419, 620–624 (2002)

    ADS  Article  Google Scholar 

  18. Mii, Y. & Taira, M. Secreted Frizzled-related proteins enhance the diffusion of Wnt ligands and expand their signaling range. Development 136, 4083–4088 (2009)

    CAS  Article  Google Scholar 

  19. Petersen, C. P. & Reddien, P. W. Polarized notum activation at wounds inhibits Wnt function to promote planarian head regeneration. Science 332, 852–855 (2011)

    ADS  CAS  Article  Google Scholar 

  20. Stephan-Dubois, F. & Gilgenkrantz, F. Transplantation et régénération chez la planaire Dendrocoelum lacteum. J. Embryol. Exp. Morphol. 9, 642–649 (1961)

    CAS  PubMed  Google Scholar 

  21. Tasaki, J. et al. ERK signaling controls blastema cell differentiation during planarian regeneration. Development 138, 2417–2427 (2011)

    CAS  Article  Google Scholar 

  22. Calow, P. Life Cycles. An Evolutionary Approach to the Physiology of Reproduction, Development and Ageing (Chapman and Hall, 1978)

    Google Scholar 

  23. Romero, R. & Baguna, J. Quantitative cellular analysis of life-cycle strategies of iteroparous and semelparous triclads. Fortschr. Zool. 36, 283–289 (1988)

    Google Scholar 

  24. Fedecka-Bruner, B. Etudes sur la régénération des organes génitaux chez la planaire Dugesia lugubris I. Régénération des testicules après destruction. Bull. Biol. Fr. Belg. 101, 255–319 (1967)

    CAS  PubMed  Google Scholar 

  25. Grasso, M. & Gardenghi, G. The involvement of cellular elements other than neoblasts in Dendrocoelum lacteum regeneration. Boll. Zool. 45, 365–368 (1978)

    Article  Google Scholar 

  26. Pearson, B. J. et al. A formaldehyde-based whole-mount in situ hybridization method for planarians. Dev. Dyn. 238, 443–450 (2009)

    Article  Google Scholar 

  27. Chong, T., Stary, J. M., Wang, Y. & Newmark, P. A. Molecular markers to characterize the hermaphroditic reproductive system of the planarian Schmidtea mediterranea. BMC Dev. Biol. 11, 69 (2011)

    Article  Google Scholar 

  28. Rouhana, L., Vieira, A. P., Roberts-Galbraith, R. H. & Newmark, P. A. PRMT5 and the role of symmetrical dimethylarginine in chromatoid bodies of planarian stem cells. Development 139, 1083–1094 (2012)

    CAS  Article  Google Scholar 

  29. Marioni, J. C., Mason, C. E., Mane, S. M., Stephens, M. & Gilad, Y. RNAseq: An assessment of technical reproducibility and comparison with gene expression arrays. Genome Res. 18, 1509–1517 (2008)

    CAS  Article  Google Scholar 

  30. Collins, J. J., III et al. Genome-wide analysis reveals a role for peptide hormones in regulating planarian germ line development. PLoS Biol. 8, e1000509 (2010)

    Article  Google Scholar 

  31. Cebria, F. & Newmark, P. A. Planarian homologues of netrin and netrin receptor are required for proper regeneration of the central nervous system and the maintenance of nervous system architecture. Development 132, 3691–3703 (2005)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank Newmark laboratory members for comments, A. Vieira for technical assistance, A. Hernandez and the W. M. Keck Center for Comparative and Functional Genomics for sequencing assistance, and the National Forest Service, Illinois Department of Natural Resources, USA, and Montgomery County (Maryland) Department of Parks, USA, for field collection permits. We also thank A. Boney, J. Brubacher, T. Chong, M. Issigonis, H. Iyer and B. Lambrus for assistance in field collections. This work was supported by National Institute of General Medicine Sciences fellowship F32GM097921 (J.M.S.). P.A.N. is an investigator of the Howard Hughes Medical Institute.

Author information

Authors and Affiliations

Authors

Contributions

Both authors contributed to the design of the experimental strategy. J.M.S. conducted all experiments, analysed the data and drafted the manuscript, which was critically reviewed and revised by P.A.N. Both authors discussed the results and commented on the final version of the manuscript.

Corresponding author

Correspondence to James M. Sikes.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-5 and Supplementary Tables 1-3. The figures demonstrate the differences in regenerative potential of P. fluviatilis tissues, the upregulation of mitotic activity following amputation in regeneration-proficient and regeneration-deficient tissues, the effects of RNAi knockdown of Wnt signaling molecules during anterior and posterior regeneration, sequence alignments, nucleotide sequences used to generate dsRNA, and tables illustrate gene upregulation assayed by RNAseq, effects of RNAi, and oligonucleotide sequences. (PDF 11196 kb)

Behaviour of PF-Beta-catenin-1 (RNAi) knockdown tail fragment, 14 days of regeneration

Anterior-directed movements occur in Pf-β-catenin1(RNAi) animals after rescued regeneration, suggesting complete and functional head regeneration. (Time lapse covers a period of 1.3 min.) (MOV 2444 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sikes, J., Newmark, P. Restoration of anterior regeneration in a planarian with limited regenerative ability. Nature 500, 77–80 (2013). https://doi.org/10.1038/nature12403

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature12403

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.

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