Axon Growth and Regeneration Methods and Protocols

Axon Growth and Regeneration: Methods and Protocols brings together a diverse set of techniques for the study of the mechanisms underlying central nervous system axon growth, consequently providing a resource that will aid in the development of repair strategies. After an introductory section, this detailed volume continues with sections focusing on axon growth in vitro, providing a range of protocols that can be used to examine intracellular signalling pathways, axonal responses to extracellular factors and methods for quantifying outgrowth. The next section provides protocols for inducing experimental injury in vivo as well as some highly promising protocols for promoting regeneration, which segues into the final section highlighting a series of protocols that can be used to monitor the extent of axon regeneration in vivo, ranging from tract tracing to in vivo imaging and functional recovery. As a book in the Methods in Molecular Biology series, chapters contain introductions 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.

Practical and reliable, Axon Growth and Regeneration: Methods and Protocols aims to serve researchers studying axon regeneration with a significant set of diverse tools, vital for moving on to the next generation of exciting new discoveries in the field.

Part I: Review

1. Axon Regeneration: What Needs to Be Overcome?

            Andrew J. Murray

Part II: Monitoring Intracellular Pathways in Growth Cones

2. Analysis of Calcium Signals in Steering Neuronal Growth Cones In Vitro

            Hiroki Akiyama and Hiroyuki Kamiguchi

3. The Use of Fluorescence Resonance Energy Transfer (FRET) to Measure Axon Growth and Guidance Related Intracellular Signaling in Live Dorsal Root Ganglia Neuronal Growth Cones

            Steven J. Tucker

Part III: Assessing Axonal Responses to Extracellular Factors In Vitro

4. Neurite Outgrowth and Growth Cone Collapse Assays to Assess Neuronal Responses to Extracellular Cues

            Andrew Kaplan, Ricardo Sanz, Gino B. Ferraro, Ricardo Alchini, and Alyson E. Fournier

5. Quantitative Assessment of Neurite Outgrowth in Mouse Retinal Explants

            Tom Buyens, Djoere Gaublomme, Inge Van Hove, Lies De Groef, and Lieve Moons

6. Growth Cone Collapse Assay

            Geoffrey M.W. Cook, Prem Jareonsettasin, and Roger J. Keynes

7. Axon Length Quantification Microfluidic Culture Platform for Growth and Regeneration Study

            Jaewon Park, Sunja Kim, Jianrong Li, and Arum Han

8. Organotypic Slice Co-Culture Systems to Study Axon Regeneration in the Dopaminergic System Ex Vivo

            Claudia Heine and Heike Franke

9. Monitoring Neuron and Astrocyte Interactions with a 3D Cell Culture System

            James B. Phillips

Part IV: Axon Injury and Regeneration In Vivo

10. Targeting Inhibitory Chondroitin Sulphate Proteoglycans to Promote Plasticity After Injury

            Jessica C.F. Kwok, Janosch P. Heller, Rong-Rong Zhao, and James W. Fawcett

11. Polymeric Biomaterials for Nerve Regeneration: Fabrication and Implantation of a Biodegradable Nerve Guide

            Wesley N. Sivak, Jacqueline M. Bliley, and Kacey G. Marra

12. A Highly Reproducible Mouse Model of Compression Spinal Cord Injury

            Suelen Adriani Marques, Fernanda Martins de  Almeida, Klauss Mostacada, and Ana Maria Blanco Martinez

13. Using Templated Agarose Scaffolds to Promote Axon Regeneration Through Sites of Spinal Cord Injury

            Jacob Koffler, Ramsey F. Samara, and Ephron S. Rosenzweig

14. In Vivo Electroporation of Adult Mouse Sensory Neurons for Studying Peripheral Axon Regeneration

            Saijilafu, Bo-Yin Zhang, and Feng-Quan Zhou

Part V: Assessing the Extent of Axon Regeneration

15. Assessing Motor Outcome and Functional Recovery Following Nerve Injury

            Ronald Deumens, Claudia Marinangeli, Ahmet Bozkurt, and Gary Anthony Brook

16. The Use of an Adeno-Associated Viral Vector for Efficient Bicistronic Expression of Two Genes in the Central Nervous System

            Thomas Haynes Hutson, Claudia Kathe, Sean Christopher Menezes, Marie-Claire Rooney, Hansruedi Bueler, and Lawrence David Falcon Moon

17. Application of Tissue Clearing and Light Sheet Fluorescence Microscopy to Assess Optic Nerve Regeneration in Unsectioned Tissues

            Xueting Luo, Benjamin Yungher, and Kevin K. Park

18. Time-Lapse In Vivo Imaging of Dorsal Root Nerve Regeneration in Mice

            Andrew Skuba, Meredith Ann Manire, Hyukmin Kim, Seung Baek Han, and Young-Jin Son

19. Using Manganese-Enhanced MRI to Assess Optic Nerve Regeneration

            Ioanna Sandvig and Axel Sandvig