Studying Newt Brain Regeneration Following Subtype Specific Neuronal Ablation

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

Abstract

The realization that neuronal injury does not result in permanent functional or cellular loss in all vertebrates has fascinated regenerative biologists. Neuronal regeneration occurs in a subset of species, including lizards, teleost fish, axolotls, and newts. One tool for studying neuronal regeneration in the adult brain is intraventricular injection of selective neuronal toxins, which leads to loss of subpopulations of neurons. To trace cells involved in the regeneration process, plasmids encoding reporter proteins can be electroporated in vivo into the cells of interest. This protocol describes methods to label the ependymoglial cells of the brain of the red spotted newt Notophthalmus viridescens and follow their response after ablation of dopaminergic neurons.

Key words

Neurogenesis In vivo electroporation 6-OHDA Dopaminergic neurons Neuronal regeneration 

Notes

Acknowledgement

This work was supported by a grant from the Swedish Research Council to MK.

References

  1. 1.
    Tanaka EM, Ferretti P (2009) Considering the evolution of regeneration in the central nervous system. Nat Rev Neurosci 10:713–723CrossRefGoogle Scholar
  2. 2.
    Kokaia Z, Martino G, Schwartz M, Lindvall O (2012) Cross-talk between neural stem cells and immune cells: the key to better brain repair? Nat Neurosci 15:1078–1087CrossRefGoogle Scholar
  3. 3.
    Zupanc GKH, Sîrbulescu RF (2011) Adult neurogenesis and neuronal regeneration in the central nervous system of teleost fish. Eur J Neurosci 34:917–929CrossRefGoogle Scholar
  4. 4.
    Lang DM, Monzón-Mayor M, Bandtlow CE, Stuermer CA (1998) Retinal axon regeneration in the lizard Gallotia galloti in the presence of CNS myelin and oligodendrocytes. Glia 23:61–74CrossRefGoogle Scholar
  5. 5.
    Okamoto M, Ohsawa H, Hayashi T et al (2007) Regeneration of retinotectal projections after optic tectum removal in adult newts. Mol Vis 13:2112–2118Google Scholar
  6. 6.
    Kroehne V, Freudenreich D, Hans S et al (2011) Regeneration of the adult zebrafish brain from neurogenic radial glia-type progenitors. Development 138:4831–4841CrossRefGoogle Scholar
  7. 7.
    Simola N, Morelli M, Carta AR (2007) The 6-hydroxydopamine model of Parkinson’s disease. Neurotox Res 11:151–167CrossRefGoogle Scholar
  8. 8.
    Parish CL, Beljajeva A, Arenas E, Simon A (2007) Midbrain dopaminergic neurogenesis and behavioural recovery in a salamander lesion-induced regeneration model. Development 134:2881–2887CrossRefGoogle Scholar
  9. 9.
    Berg DA, Kirkham M, Beljajeva A et al (2010) Efficient regeneration by activation of neurogenesis in homeostatically quiescent regions of the adult vertebrate brain. Development 137:4127–4134CrossRefGoogle Scholar
  10. 10.
    Haas K, Jensen K, Sin WC et al (2002) Targeted electroporation in Xenopus tadpoles in vivo—from single cells to the entire brain. Differentiation 70:148–154CrossRefGoogle Scholar
  11. 11.
    Echeverri K, Tanaka EM (2002) Ectoderm to mesoderm lineage switching during axolotl tail regeneration. Science 298:1993–1996CrossRefGoogle Scholar
  12. 12.
    Barnabé-Heider F, Meletis K, Eriksson M et al (2008) Genetic manipulation of adult mouse neurogenic niches by in vivo electroporation. Nat Methods 5:189–196CrossRefGoogle Scholar
  13. 13.
    Köster RW, Fraser SE (2001) Tracing transgene expression in living zebrafish embryos. Dev Biol 15:329–346CrossRefGoogle Scholar
  14. 14.
    Kawakami K (2007) Tol2: a versatile gene transfer vector in vertebrates. Genome Biol 8(Suppl 1):S7CrossRefGoogle Scholar
  15. 15.
    Sandoval-Guzmán T, Wang H, Khattak S et al (2014) Fundamental differences in dedifferentiation and stem cell recruitment during skeletal muscle regeneration in two salamander species. Cell Stem Cell 14:174–187CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Cell and Molecular BiologyKarolinska InstitutetStockholmSweden

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