Axon Degeneration: Methods and Protocols

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

            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

            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 López-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