Main description:

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:

1 Salamanders as key models for development and regeneration research

Malcolm Maden

Part I: LABORATORY COLONY HUSBANDRY

2 Establishing a new research axolotl colony

Anastasia S. Yandulskaya and James R. Monaghan

3 Health monitoring for laboratory salamanders

Marcus J. Crim and Marcia L. Hart

4 Husbandry, captive breeding and field survey of Chinese giant salamander (Andrias davidianus)

Wansheng Jiang, Haifeng Tian, and Lu Zhang

Part II: TRADITIONAL MOLECULAR TECHNIQUES

5 Whole mount in situ hybridization (WISH) for salamander embryos

and larvae

Sruthi Purushothaman and Ashley W. Seifert

6 Hybridization chain reaction fluorescence in situ hybridization

(HCR-FISH) in Ambystoma mexicanum tissue

Alex M. Lovely, Timothy J. Duerr, David F. Stein, Evan T. Mun, and James R. Monaghan

7 Ethyl cinnamate-based tissue clearing strategies Wouter Masselink and Elly M. Tanaka

8 Induction and characterization of cellular senescence in salamanders

Qinghao Yu, Hannah E. Walters, and Maximina H. Yun

9 Methods for studying appendicular skeletal biology in axolotls

Camilo Riquelme-Guzman and Tatiana Sandoval-Guzman

10 Fluorescence in situ hybridization of DNA-probes on mitotic

chromosomes of the Mexican axolotl

Melissa Keinath and Vladimir Timoshevskiy

11 The use of small molecules to dissect developmental and regenerative

processes in axolotls

Stephane Roy

12 COMET Assay for detection of DNA damage during axolotl tail

regeneration

Belfran Carbonell, Jennifer Alvarez, Gloria A. Santa-Gonzalez and

Jean Paul Delgado

Part III: EXPERIMENTAL MANIPULATIONS AND SURGERIES

13 In vivo and Ex vivo View of Newt Lens Regeneration

Georgios Tsissios, Anthony Sallese, Weihao Chen, Alyssa Miller, Hui Wang

and Katia Del Rio-Tsonis

14 Bead implantation and delivery of exogenous growth factors

Rena Kashimoto, Saya Furukawa, Sakiya Yamamoto, and Akira Satoh

15 The Accessory Limb Model regenerative assay and its derivatives

Michael Raymond and Catherine D. McCusker

16 Embryonic Tissue and Blastema Transplantations

Maritta Schuez, Thomas Kurth, Joshua D. Currie, and Tatiana

Sandoval-Guzman

17 Retinoic acid-induced limb duplications Malcolm Maden and Trey Polvadore

18 Isolation and characterization of peritoneal macrophages from salamanders Anthony Sallese, Georgios Tsissios, Jose Raul Perez-Estrada, Arielle Martinez and Katia Del Rio-Tsonis

Part IV: BIOINFORMATICS AND GENOMICS

19 Navigation and use of custom tracks within the axolotl genome browser

Sergej Nowoshilow and Elly M. Tanaka

20 Chromosome Conformation Capture for large genomes

Akane Kawaguchi and Elly Tanaka

Part V: TRANSGENICS AND LINEAGE-TRACING

21 Axolotl transgenesis via injection of I-SceI meganuclease or Tol2

transposon system

Maritta Schuez and Tatiana Sandoval-Guzman

22 A practical guide for CRISPR-Cas9-induced mutations in axolotls

Konstantinos Sousounis, Katharine Courtemanche, Jessica L. Whited

23 Applying a knock-in strategy to create reporter tagged knockout alleles in axolotls Liqun Wang, Yan-Yun Zeng, Yanmei Liu and Ji-Feng Fei

24 Baculovirus production and infection in axolotls

Prayag Murawala, Catarina R. Oliveira, Helena Okulski,

Maximina H. Yun, and Elly M. Tanaka

25 Cell dissociation techniques in salamanders

Gabriela Johnson, Nadjib Dastagir, Zachary Beal, Andrew Hart, and

James Godwin

Part VI : PHYSIOLOGICAL AND ORGANISMAL TECHNIQUES

26 Axolotl metabolism: measuring metabolic energy

Moshe Khurgel

27 Artificial insemination in axolotls

Yuka Taniguchi-Sugiura and Elly Tanaka

28 Screening salamanders for symbionts

Elli Vickers and Ryan Kerney

29 Assessing leukocyte profiles of salamanders and other amphibians:

a herpetologists' guide

Andrew K. Davis and John C. Maerz

Part VII: EPILOGUE

30 Best practices to promote data utility and recycling by the salamander

community

Garrett S. Dunlap and Nicholas D. Leigh

31 Now that we got there, what next?

Elly Tanaka