Salamanders in Regeneration Research Methods and Protocols

About this book

This detailed volume focuses on best practices and conditions for maintaining the most commonly used salamander species in the laboratory. Salamanders in Regeneration ResearchMethods and Protocols guides readers through experimental manipulations in vivo and in vitro, respectively. With methods on targeting a wide variety of structures, ranging from the limb to the heart and to the brain, and methods for studying genetically modified organisms and tools for mining in the genomic databases. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Salamanders in Regeneration Research: Methods and Protocols provides a comprehensive collection of methods chapters.

Contents

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 Andras Simon
5. Maintaining Plethodontid Salamanders in the Laboratory for Regeneration Studies Claudia Marcela Arenas G, Andrea Gomez-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-Francois Denis, Fadi Sader, Patrizia Ferretti, and Stephane Roy
16. Isolation and Culture of Neurospheres from the Adult Newt Brain L Shahul Hameed, Andras 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

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