Skip to main content

Maintaining Eastern Newts (Notophthalmus viridescens) for Regeneration Research

  • Protocol
  • First Online:
Salamanders in Regeneration Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1290))

Abstract

The adult Eastern newt, Notophthalmus viridescens, has long served as a model for appendage as well as heart muscle regeneration studies. Newt tissues include all major cell types known in other vertebrates and mammals, including bone, cartilage, tendon, muscle, nerves, dermis, and epidermis. Therefore, these aquatic salamanders make an excellent model for studying the regeneration of complex tissues. Regeneration of adult tissues requires the integration of new tissues with preexisting tissues to form a functioning unit through a process that is not yet well understood. Scale is also an issue, because the regenerating tissues or structures are magnitudes larger than their embryonic counterparts during development, and therefore, it is likely that different physics and mechanics apply. Regardless, regeneration recapitulates to some degree developmental processes. In this chapter, we will describe basic methods for maintaining adult Eastern newts in the laboratory for the study of regeneration. To determine similarities and differences between development and regeneration at the cellular and molecular level, there is also a need for embryonic newt tissue. We therefore also outline a relatively simple way to produce and raise newt embryos in the laboratory.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Duellman WE, Trueb L (1986) Biology of amphibians. McGraw-Hill Book Company, New York, NY

    Google Scholar 

  2. Petranka JW (1996) Salamanders of the United States and Canada. Smithsonian Institution Press, Washington, DC

    Google Scholar 

  3. Spallanzani L (1769) An essay on animal reproductions. Translated from Italian by M. Maty. Printed for T. Becket, and P. A. de Hondt, in the Strand, London. Available through UMI Books on Demand

    Google Scholar 

  4. Wallace H (1981) Vertebrate Limb Regeneration. J Wiley and Sons Ltd., Toronto, ON

    Google Scholar 

  5. Nye HL, Cameron JA, Chernoff EA, Stocum DL (2003) Regeneration of the urodele limb: a review. Dev Dyn 226:280–294

    Article  Google Scholar 

  6. Eguchi G, Eguchi Y, Nakamura K, Yadav MC, Lillan JL, Tsonis PA (2011) Regenerative capacity in newts is not altered by repeated regeneration and ageing. Nat Commun 2:384. doi:10.1038/ncomms1389

    Article  Google Scholar 

  7. Eguchi G, Itoh Y (1982) Regeneration of the lens as a phenomenon of cellular transdifferentiation: regulability of the differentiated state of the vertebrate pigment epithelial cell. Trans Ophthalmol Soc U K 3:380–384

    Google Scholar 

  8. Oberpriller J, Oberpriller JC (1971) Mitosis in adult newt ventricle. J Cell Biol 49:560–563

    Article  CAS  Google Scholar 

  9. Bettencourt-Dias M, Mittnacht S, Brockes JP (2003) Heterogeneous proliferative potential in regenerative adult newt cardiomyocytes. J Cell Sci 116:4001–4009

    Article  CAS  Google Scholar 

  10. Calve S, Odelberg SJ, Simon H-G (2010) A transitional extracellular matrix instructs cell behavior during muscle regeneration. Dev Biol 344: 259-271

    Google Scholar 

  11. Calve S and Simon H-G (2011) High resolution 3D imaging: Evidence for cell cycle reentry in regenerating skeletal muscle. Developmental Dynamics 240:1233-1239

    Google Scholar 

  12. Lo DC, Allen F, Brockes JP (1993) Reversal of muscle differentiation during urodele limb regeneration. Proc Natl Acad Sci U S A 90:7230–7234

    Article  CAS  Google Scholar 

  13. Sandoval-Guzmán T, Wang H, Khattak S, Schuez M, Roensch K, Nacu E, Tazaki A, Joven A, Tanaka EM, Simon A (2014) Fundamental differences in dedifferentiation and stem cell recruitment during skeletal muscle regeneration in two salamander species. Cell Stem Cell 14:174–187

    Article  Google Scholar 

  14. Piatt J (1955) Regeneration of the spinal cord in the salamander. J Exp Zool 129:177–207

    Article  Google Scholar 

  15. Davis BM, Ayers JL, Koran L, Carlson J, Anderson MC, Simpson SB Jr (1990) Time course of salamander spinal cord regeneration and recovery of swimming: HRP retrograde pathway tracing and kinematic analysis. Exp Neurol 108:198–213

    Article  CAS  Google Scholar 

  16. Okamoto M, Ohsawa H, Hayashi T, Owaribe K, Tsonis PA (2007) Regeneration of retinotectal projections after optic tectum removal in adult newts. Mol Vis 13:2112–2118

    Google Scholar 

  17. Berg DA, Kirkham M, Beljajeva A, Knapp D, Habermann B, Ryge J, Tanaka EM, Simon A (2010) Efficient regeneration by activation of neurogenesis in homeostatically quiescent regions of the adult vertebrate brain. Development 137:4127–4134

    Article  CAS  Google Scholar 

  18. Khan PA, Liversage RA (1995) Development of Notophthalmus viridescens embryos. Dev Growth Differ 37:529–537

    Article  Google Scholar 

  19. Khan PA, Liversage RA (1995) Spawning of Notophthalmus viridescens embryos. Herpetol Rev 26:95–96

    Google Scholar 

  20. Yatsu-Yamashita M, Gilhen J, Russell RW, Krysko KL, Melaun C, Kurz A, Kauferstein S, Kordis D, Mebs D (2012) Variability of tetrodotoxin and of its analogues in the red-spotted newt, Notophthalmus viridescens (Amphibia: Urodela: Salamandridae). Toxicon 59:257–264

    Article  Google Scholar 

Download references

Acknowledgments

We would like to acknowledge both current and former researchers in our respective laboratories who helped develop the protocols described in this chapter. These individuals include Paul Khan, Barbara Linkhart, Claudia Guzman, Sarah Calve, Sarah Mercer, Donald Atkinson, Vladimir Vinarsky, Tamara Stevenson, David Kent, and Katherine Zukor. We would also like to acknowledge several of our colleagues who provided us with invaluable advice when we were just beginning our studies on regeneration using this remarkable animal, including Cliff Tabin, Mark Keating, Jeremy Brockes, David Stocum, Roy Tassava, Panagiotis Tsonis, and Anthony Mescher.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hans-Georg Simon Ph.D. .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Simon, HG., Odelberg, S. (2015). Maintaining Eastern Newts (Notophthalmus viridescens) for Regeneration Research. In: Kumar, A., Simon, A. (eds) Salamanders in Regeneration Research. Methods in Molecular Biology, vol 1290. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2495-0_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-2495-0_2

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2494-3

  • Online ISBN: 978-1-4939-2495-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics