Salamanders Methods and Protocols

About this book

This volume provides a comprehensive reference for researchers aiming to bring new techniques and approaches to their scientific research using urodeles. Chapters are authored by leaders in the field and meant to guide readers through laboratory colony husbandry, traditional molecular techniques, experimental manipulation and surgeries, bioinformatics and genomics, transgenics and lineage-tracing, and physiological and organismal techniques. In addition to laboratory methods, this volume highlights techniques developed for field studies and work with wild-caught animals. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and state-of-the-art, Methods in Salamander Research aims to be a practical guide for researchers interested in deploying new methodologies in their lab or in the field.

Contents

- Salamanders as key models for development and regeneration research
- Establishing a new research axolotl colony
- Health monitoring for laboratory salamanders
- Husbandry, captive breeding and field survey of Chinese giant salamander (Andrias davidianus)
- Whole mount in situ hybridization (WISH) for salamander embryos and larvae
- Hybridization chain reaction fluorescence in situ hybridization (HCR-FISH) in Ambystoma mexicanum tissue
- Ethyl cinnamate-based tissue clearing strategies
- Induction and characterization of cellular senescence in salamanders
- Methods for studying appendicular skeletal biology in axolotls
- Fluorescence in situ hybridization of DNA-probes on mitotic chromosomes of the Mexican axolotl
- The use of small molecules to dissect developmental and regenerative processes in axolotls
- COMET Assay for detection of DNA damage during axolotl tail regeneration
- In vivo and Ex vivo View of Newt Lens Regeneration
- Bead implantation and delivery of exogenous growth factors
- The Accessory Limb Model regenerative assay and its derivatives
- Embryonic Tissue and Blastema Transplantations
- Retinoic acid-induced limb duplications
- Isolation and characterization of peritoneal macrophages from salamanders
- Navigation and use of custom tracks within the axolotl genome browser
- Chromosome Conformation Capture for large genomes
- Axolotl transgenesis via injection of I-SceI meganuclease or Tol2 transposon system
- A practical guide for CRISPR-Cas9-induced mutations in axolotls
- Applying a knock-in strategy to create reporter tagged knockout alleles in axolotls
- Baculovirus production and infection in axolotls
- Cell dissociation techniques in salamanders
- Axolotl metabolism: measuring metabolic energy
- Artificial insemination in axolotls
- Screening salamanders for symbionts
- Assessing leukocyte profiles of salamanders and other amphibians: a herpetologists' guide
- Best practices to promote data utility and recycling by the salamander community
- Now that we got there, what's next?.

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