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This detailed volume focuses on best practices and conditions for maintaining the most commonly used salamander species in the laboratory. Salamanders in Regeneration Research: Methods 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.
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Table of Contents

Part I: Salamanders1. Variation in Salamanders: An Essay on Genomes, Development, and EvolutionJeremy P Brockes 2. Maintaining Eastern Newts (Notophthalmus viridescens) for Regeneration ResearchHans-Georg Simon and Shannon Odelberg3. Housing and Maintenance of Ambystoma mexicanum, the Mexican AxolotlJohanna 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 StudiesClaudia Marcela Arenas G, Andrea Gomez-Molina, and Jean Paul Delgado Part II: Experimental Manipulation in Salamanders 6. Newt Lens Transdifferentiation: From Lentectomy to Immuno-FISHNobuyasu. Maki 7. Studying Newt Brain Regeneration Following Subtype Specific Neuronal AblationMatthew 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 Satoh9. High-efficiency Electroporation of the Spinal Cord in Larval AxolotlAida Rodrigo-Albors and Elly M. Tanaka 10. Pseudotyped Retroviruses for Infecting AxolotlTzu-Hsing Kuo and Jessica L. Whited11. Thyroxine Induced Metamorphosis in the Axolotl (Ambystoma mexicanum)Peggy S. Coots and Ashley W. Seifert 12. Generation of Aneurogenic Larvae by Parabiosis of Salamander EmbryosAnoop Kumar and Jean Paul Delgado 13. In vivo Modulation and Quantification of microRNAs During Axolotl Tail RegenerationJami R. Erickson and Karen EcheverriPart III: Salamander Cells in Culture14. 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 CellsJean-Francois Denis, Fadi Sader, Patrizia Ferretti, and Stephane Roy 16. Isolation and Culture of Neurospheres from the Adult Newt BrainL Shahul Hameed, Andras Simon 17. Methods for Axolotl Blood Collection, Intravenous Injection and Efficient Leukocyte Isolation from Peripheral Blood and the Regenerating LimbRyan J. Debuque and James W. Godwin 18. Assessing Cardiomyocyte Proliferative Capacity in the Newt Heart and Primary CultureHans-Georg Simon and Shannon Odelberg 19. Long Term Organ Cultures of Newt HeartsTanja 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 Salamanders21. Transgenesis in Axolotl (Ambystoma mexicanum)Shahryar Khattak and Elly M Tanaka 22. Generating and Identifying Axolotls with Targeted Mutations using Cas9 RNA-guided NucleaseG Parker Flowers and Craig M Crews 23. Gene Manipulation for Regenerative Studies using the Iberian Ribbed Newt, Pleurodeles waltlToshinori Hayashi and Takashi Takeuchi Part V: Gene Expression24. Transcriptomics Using AxolotlsS. 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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