Xenopus pp 133-146 | Cite as

Conditional Chemogenetic Ablation of Photoreceptor Cells in Xenopus Retina

  • Albert Chesneau
  • Odile Bronchain
  • Muriel PerronEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1865)


Xenopus is an attractive model system for regeneration studies, as it exhibits an extraordinary regenerative capacity compared to mammals. It is commonly used to study body part regeneration following amputation, for instance of the limb, the tail, or the retina. Models with more subtle injuries are also needed for human degenerative disease modeling, allowing for the study of stem cell recruitment for the regeneration of a given cellular subtype. We present here a model to ablate photoreceptor cells in the Xenopus retina. This method is based on the nitroreductase/metronidazole (NTR/MTZ) system, a combination of chemical and genetic tools, allowing for the conditional ablation of targeted cells. This type of approach establishes Xenopus as a powerful model to study cellular regeneration and stem cell regulation.

Key words

Xenopus Cell ablation Regeneration Retina Photoreceptors NTR-MTZ 



We thank B.E. Knox for providing the Rhodopsin promoter plasmid. Our lab is supported by grants from the Fondation pour la Recherche Médicale (FRM), Association Retina France and Fondation Valentin Haüy.


  1. 1.
    Li J, Zhang S, Amaya E (2016) The cellular and molecular mechanisms of tissue repair and regeneration as revealed by studies in Xenopus. Regeneration 3:198–208. Scholar
  2. 2.
    Tseng a-S, Levin M (2008) Tail regeneration in Xenopus laevis as a model for understanding tissue repair. J Dent Res 87:806–816. Scholar
  3. 3.
    Beck CW, Izpisúa Belmonte JC, Christen B (2009) Beyond early development: Xenopus as an emerging model for the study of regenerative mechanisms. Dev Dyn 238:1226–1248. Scholar
  4. 4.
    Slack JMW, Lin G, Chen Y (2008) The Xenopus tadpole: a new model for regeneration research. Cell Mol Life Sci 65:54–63. Scholar
  5. 5.
    Yoshii C, Ueda Y, Okamoto M, Araki M (2007) Neural retinal regeneration in the anuran amphibian Xenopus laevis post-metamorphosis: transdifferentiation of retinal pigmented epithelium regenerates the neural retina. Dev Biol 303:45–56. Scholar
  6. 6.
    Araki M (2007) Regeneration of the amphibian retina: role of tissue interaction and related signaling molecules on RPE transdifferentiation. Develop Growth Differ 49:109–120. Scholar
  7. 7.
    Araki M (2014) A novel mode of retinal regeneration: the merit of a new Xenopus model. Neural Regen Res 9:2125–2127. Scholar
  8. 8.
    Chiba C (2014) The retinal pigment epithelium: an important player of retinal disorders and regeneration. Exp Eye Res 123:107–114. Scholar
  9. 9.
    Miyake A, Araki M (2014) Retinal stem/progenitor cells in the ciliary marginal zone complete retinal regeneration: a study of retinal regeneration in a novel animal model. Dev Neurobiol. Scholar
  10. 10.
    Ail D, Perron M (2017) Retinal degeneration and regeneration—lessons from fishes and amphibians. Curr Pathobiol Rep 5:67–78. Scholar
  11. 11.
    Choi RY, Engbretson GA, Solessio EC et al (2011) Cone degeneration following rod ablation in a reversible model of retinal degeneration. Invest Ophthalmol Vis Sci 52:364–373. Scholar
  12. 12.
    Hamm LM, Tam BM, Moritz OL (2009) Controlled rod cell ablation in transgenic Xenopus laevis. Invest Ophthalmol Vis Sci 50:885–892. Scholar
  13. 13.
    Langhe R, Chesneau A, Colozza G et al (2017) Müller glial cell reactivation in Xenopus models of retinal degeneration. Glia 65:1333–1349. Scholar
  14. 14.
    Curado S, Anderson RM, Jungblut B et al (2007) Conditional targeted cell ablation in zebrafish: a new tool for regeneration studies. Dev Dyn 236:1025–1035. Scholar
  15. 15.
    Curado S, Stainier D (2008) Nitroreductase-mediated cell/tissue ablation in zebrafish: a spatially and temporally controlled ablation method with applications in developmental and regeneration studies. Nat Protoc 3:948–954. Scholar
  16. 16.
    White DT, Mumm JS (2013) The nitroreductase system of inducible targeted ablation facilitates cell-specific regenerative studies in zebrafish. Methods 62:232–240. Scholar
  17. 17.
    White DT, Sengupta S, Saxena MT et al (2017) Immunomodulation-accelerated neuronal regeneration following selective rod photoreceptor cell ablation in the zebrafish retina. Proc Natl Acad Sci 114:E3719–E3728. Scholar
  18. 18.
    Pipalia TG, Koth J, Roy SD et al (2016) Cellular dynamics of regeneration reveals role of two distinct Pax7 stem cell populations in larval zebrafish muscle repair. Dis Model Mech 9:671–684. Scholar
  19. 19.
    Ariga J, Walker SL, Mumm JS (2010) Multicolor time-lapse imaging of transgenic zebrafish: visualizing retinal stem cells activated by targeted neuronal cell ablation. J Vis Exp.
  20. 20.
    Ohnmacht J, Yang Y, Maurer GW et al (2016) Spinal motor neurons are regenerated after mechanical lesion and genetic ablation in larval zebrafish. Development 143:1464–1474. Scholar
  21. 21.
    Oosterhof N, Kuil LE, van Ham TJ (2017) Microglial activation by genetically targeted conditional neuronal ablation in the zebrafish. Meth Mol Biol (Clifton, NJ) 1559:377–390CrossRefGoogle Scholar
  22. 22.
    White YAR, Woods DC, Wood AW (2011) A transgenic zebrafish model of targeted oocyte ablation and de novo oogenesis. Dev Dyn 240:1929–1937. Scholar
  23. 23.
    Fraser B, DuVal MG, Wang H, Allison WT (2013) Regeneration of cone photoreceptors when cell ablation is primarily restricted to a particular cone subtype. PLoS One 8:e55410. Scholar
  24. 24.
    Montgomery JE, Parsons MJ, Hyde DR (2010) A novel model of retinal ablation demonstrates that the extent of rod cell death regulates the origin of the regenerated zebrafish rod photoreceptors. J Comp Neurol 518:800–814. Scholar
  25. 25.
    Kaya F, Mannioui A, Chesneau A et al (2012) Live imaging of targeted cell ablation in Xenopus: a new model to study demyelination and repair. J Neurosci 32:12885–12895. Scholar
  26. 26.
    Mannioui A, Vauzanges Q, Fini JB et al (2017) The Xenopus tadpole: an in vivo model to screen drugs favoring remyelination. Mult Scler J. Scholar
  27. 27.
    Knox BE, Schlueter C, Sanger BM et al (1998) Transgene expression in Xenopus rods. FEBS Lett 423:117–121CrossRefGoogle Scholar
  28. 28.
    Ishibashi S, Kroll KL, Amaya E (2012) Generating transgenic frog embryos by restriction enzyme mediated integration (REMI). Methods Mol Biol 917:185–203. Scholar
  29. 29.
    Chesneau A, Sachs LM, Chai N et al (2008) Transgenesis procedures in Xenopus. Biol Cell 100:503–521. Scholar
  30. 30.
    Kroll KL, Amaya E (1996) Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation. Development 122:3173–3183PubMedGoogle Scholar
  31. 31.
    Nieuwkoop P, Faber J (1994) Normal table of Xenopus laevis. In: Garland, TXGoogle Scholar
  32. 32.
    Nye HLD, Cameron JA (2005) Strategies to reduce variation in Xenopus regeneration studies. Dev Dyn 234:151–158. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Albert Chesneau
    • 1
  • Odile Bronchain
    • 1
  • Muriel Perron
    • 1
    Email author
  1. 1.Paris-Saclay Institute of NeuroscienceCNRS, Univ Paris Sud, Université Paris-SaclayOrsay CedexFrance

Personalised recommendations