Salamanders in Regeneration Research: Methods and Protocols

Part I: Salamanders

1. Variation in Salamanders: An Essay on Genomes, Development, and Evolution

Jeremy P Brockes

2. Maintaining Eastern Newts (Notophthalmus viridescens) for Regeneration Research

Hans-Georg Simon and Shannon Odelberg

3. Housing and Maintenance of Ambystoma mexicanum, the Mexican Axolotl

Johanna E. Farkas and James R. Monaghan

4. Husbandry of Spanish Ribbed Newts (Pleurodeles waltl)

Alberto Joven, Matthew Kirkham, and András Simon

5. Maintaining Plethodontid Salamanders in the Laboratory for Regeneration Studies

Claudia Marcela Arenas G, Andrea Gómez-Molina, and Jean Paul Delgado

Part II: Experimental Manipulation in Salamanders

6. Newt Lens Transdifferentiation: From Lentectomy to Immuno-FISH

Nobuyasu. Maki

7. Studying Newt Brain Regeneration Following Subtype Specific Neuronal Ablation

Matthew Kirkham and Alberto Joven

8. The Accessory Limb Model: An Alternative Experimental System of Limb Regeneration

Tetsuya Endo, David M. Gardiner, Aki Makanae, and Akira Satoh

9. High-efficiency Electroporation of the Spinal Cord in Larval Axolotl

Aida Rodrigo-Albors and Elly M. Tanaka

10. Pseudotyped Retroviruses for Infecting Axolotl

Tzu-Hsing Kuo and Jessica L. Whited

11. Thyroxine Induced Metamorphosis in the Axolotl (Ambystoma mexicanum)

Peggy S. Coots and Ashley W. Seifert

12. Generation of Aneurogenic Larvae by Parabiosis of Salamander Embryos

Anoop Kumar and Jean Paul Delgado

13. In vivo Modulation and Quantification of microRNAs During Axolotl Tail Regeneration

Jami R. Erickson and Karen Echeverri

Part III: Salamander Cells in Culture

14. Derivation and Long-Term Culture of Cells from Newt Adult Limbs and Limb Blastemas

Patrizia Ferretti and Anoop Kumar

15. Culture and Transfection of Axolotl Cells

Jean-François Denis, Fadi Sader, Patrizia Ferretti, and Stéphane Roy

16. Isolation and Culture of Neurospheres from the Adult Newt Brain

L Shahul Hameed, András Simon

17. Methods for Axolotl Blood Collection, Intravenous Injection and Efficient Leukocyte Isolation from Peripheral Blood and the Regenerating Limb

Ryan J. Debuque and James W. Godwin

18. Assessing Cardiomyocyte Proliferative Capacity in the Newt Heart and Primary Culture

Hans-Georg Simon and Shannon Odelberg

19. Long Term Organ Cultures of Newt Hearts

Tanja Piatkowski and Thomas Braun
20. In vitro Preparation of Newt Inner Ear Sensory Epithelia as a Model for Repair and Regeneration
Ruth R Taylor

Part IV Transgenesis in Salamanders

21. Transgenesis in Axolotl (Ambystoma mexicanum)

Shahryar Khattak and Elly M Tanaka

22. Generating and Identifying Axolotls with Targeted Mutations using Cas9 RNA-guided Nuclease

G Parker Flowers and Craig M Crews

23. Gene Manipulation for Regenerative Studies using the Iberian Ribbed Newt, Pleurodeles waltl

Toshinori Hayashi and Takashi Takeuchi

Part V: Gene Expression

24. Transcriptomics Using Axolotls

S. Randal Voss, Anthony Athippozhy, and M. Ryan Woodcock

25. Sal-Site: Research Resources for The Mexican Axolotl

Nour W Al Haj Baddar, M Ryan Woodcock, Shivam Khatri, D. Kevin Kump, and S. Randal Voss
26. Data Mining in Newt-omics, the Repository for Omics Data from the Newt
Mario Looso and Thomas Braun