Main description:

This book is a collection of classical as well as innovative methods used to investigate axon degeneration with a particular focus on addressing the common challenges encountered while performing these procedures. Particular attention is devoted to the study of axon loss in several model organisms, as each poses unique challenges and provides powerful advantages. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Axon Degeneration: Methods and Protocols is an ideal guide for facilitating the application and further development of these protocols, which will help the scientific community tackle important questions regarding axon degeneration.
Chapters 2, 3, and 20 are available Open Access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Contents:

Part I: Introduction

1. Axon Degeneration: Which Method to Choose?

Michael P. Coleman

Part II: In Vitro Assays

2. Axon Degeneration Assays in Superior Cervical Ganglion Explant Cultures

Andrea Loreto and Jonathan Gilley

3. Microinjection of Superior Cervical Ganglion Neurons for Studying Axon Degeneration

Jonathan Gilley and Andrea Loreto

4. Assessing Axonal Degeneration in Embryonic Dorsal Root Ganglion Neurons In Vitro

Jung Eun Shin and Yongcheol Cho

5. Viral Transduction of DRG Neurons

Yo Sasaki

6. Planning and Analysis of Axon Degeneration Screening Experiments

Lyndah Lovell, Jack Bramley, and William Buchser

7. A Microfluidic Culture Platform to Assess Axon Degeneration

Yu Yong, Christopher Hughes, and Christopher Deppmann

8. A Schwann Cell-Neuron Co-Culture System to Study Neuron-Glia Interaction during Axon Degeneration

Elisabetta Babetto

9. Establishing Myelinating Co-Cultures Using Human iPSC-Derived Sensory Neurons to Investigate Axonal Degeneration and Demyelination

Alex Clark

Part III: Ex Vivo Assays

10. Preparation of Organotypic Hippocampal Slice Cultures for the Study of CNS Disease and Damage

Claire S. Durrant

11. Organotypic Culture Assay for Neuromuscular Synaptic Degeneration and Function

Kosala N. Dissanayake, Robert Chang-Chih Chou, Rosalind Brown, and Richard R. Ribchester

12. Intracellular Recordings of Postsynaptic Voltage Responses at the Drosophila Neuromuscular Junction

Vera Valakh and Johanna G. Flyer-Adams

13. Ex Vivo Studies of Optic Nerve Axon Electrophysiology

Chinthasagar Bastian, Sylvain Brunet, and Selva Baltan

14. Ex Vivo Analysis of Axonal Degeneration Using Sciatic and Optic Nerve Preparations

Rodrigo Lopez-Leal and Felipe A. Court

15. Measuring Bioenergetic Signatures of Peripheral Nerve Segments by Extracellular Flux Analysis

Bogdan Beirowski

Part IV: In Vivo Assays

16. Detection of Neutrophils in the Sciatic Nerve Following Peripheral Nerve Injury

Jon P. Niemi, Jane A. Lindborg, and Richard E. Zigmond

17. Whole-Tissue Immunolabeling and 3D Fluorescence Imaging to Visualize Axon Degeneration in the Intact, Unsectioned Mouse Tissues

Ying Cao and Jing Yang

18. Transmission Electron Microscopy and Morphometry of the CNS White Matter

Julia M. Edgar, Rebecca Sherrard Smith, and Ian D. Duncan

19. In Vivo Calcium Imaging During Axon Degeneration in Zebrafish

Mauricio E. Vargas

20. In Vivo Imaging of Anterograde and Retrograde Axonal Transport in Rodent Peripheral Nerves

James N. Sleigh, Andrew P. Tosolini, and Giampietro Schiavo

21. In Vivo Visualization of Moving Synaptic Cargo Complexes within Drosophila Larval Segmental Axons

Rupkatha Banerjee, Joseph A. White II, and Shermali Gunawardena

22. A Chemotherapy-Induced Peripheral Neuropathy Model in Drosophila melanogaster

Martha R.C. Bhattacharya

23. Models of Axon Degeneration in Drosophila Larvae

E.J. Brace and Aaron DiAntonio

24. In Vivo Analysis of Glial Immune Responses to Axon Degeneration in Drosophila melanogaster

Mary A. Logan and Sean D. Speese