Main description:

This volume collects research methodologies for both basic and translational studies of myelin authored by experts in the field. Chapters in this book are divided into three parts: Part One focuses on visualizing myelin and studying it on a biochemical level; Part Two looks at cell culture systems used to uncover key mechanisms that regulate formation and maintenance of myelin; and Part Three discusses modeling disorders of myelin in both CNS and PNS, and therapeutic strategies for targeting these disorders. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Myelin: Methods and Protocols is a valuable resource for physiologists, molecular biologists, anatomists, cell and developmental biologists, and physician-scientists working in the field of myelin biology and the disorders associated with it.

Contents:

Preface

Table of Contents

Contributing Authors

Part I Imaging Myelin and Biochemical Analysis

1. Electron Microscopy for the Analysis of Peripheral Nerve Myelin

Cinzia Ferri, Angelo Quattrini, and Maurizio D'Antonio

2. Immunohistochemical Analysis of Myelin Structures

M. Thetiot, S.A. Freeman, and A. Desmazieres

3. Visualization and Time-Lapse Microscopy of Myelinating Glia In Vivo in Zebrafish

Stavros Vagionitis and Tim Czopka

4. Lipidomics Profiling of Myelin

Chunyan Wang, Juan Pablo Palavicini, and Xianlin Han

5. Mapping Lipid Distribution in Rat Sciatic Nerve Using Imaging Mass Spectrometry

Roberto Fernandez, Jone Garate, Beatriz Abad, Begona Ochoa, and Jose A. Fernandez

6. Proteomic Analysis of Myelin Composition

Akihiro Ishii, David Han, and Rashmi Bansal

Part II Cell Culture Systems

7. Isolation and Purification of Primary Rodent Schwann Cells

Marta Palomo Irigoyen, Miguel Tamayo Caro, Encarnacion Perez Andres, Adrian Barreira Manrique, Marta Varela Rey, and Ashwin Woodhoo

8. Isolation, Expansion, and Maturation of Oligodendrocyte Lineage Cells Obtained from Rat Neonatal Brain and Optic Nerve

Maria Victoria Sanchez-Gomez, Mari Paz Serrano, Elena Alberdi, Fernando Perez-Cerda, and Carlos Matute

9. DRG Neuron/Schwann Cells Myelinating Co-Cultures

Carla Taveggia and Alessandra Bolino

10. Neuron/Oligodendrocyte Myelination Co-Culture

Yi Pang, Kimberly Simpson, Jose Javier Miguel-Hidalgo, and Renate Savich

11. Organotypic Slice Cultures to Study Oligodendrocyte Proliferation, Fate, and Myelination

Mohammad Amin Sherafat, Robert A. Hill, and Akiko Nishiyama

12. Modeling PNS and CNS Myelination Using Microfluidic Chambers

Adrien Vaquie, Alizee Sauvain, and Claire Jacob

13. Modeling CNS Myelination Using Micropillar Arrays

Nan-Xing Huang, Yun-An A. Shen, and Feng Mei

14. A Central Nervous System Axonal Myelination Assay for High-Throughput Screening

Karen Lariosa-Willingham and Dmitri Leonoudakis

15. Analyzing Autophagic Flux in Nerve Cultures

Miguel Tamayo Caro, Marta Palomo Irigoyen, Encarnacion Perez Andres, Adrian Barreira Manrique, Marta Varela Rey, and Ashwin Woodhoo

16. Subcellular Optogenetic Stimulation Platform for Studying Activity-Dependent Axon Myelination In Vitro

Agata Blasiak, Sudip Nag, and In Hong Yang

Part III Modeling Demyelinating Disorders and In Vivo Targeting Strategies

17. Active Induction of Experimental Autoimmune Encephalomyelitis (EAE) with MOD35-55 in the Mouse

Mercedes Giralt, Amalia Molinero, and Juan Hidalgo

18. Experimental Demyelination and Remyelination of Murine Spinal Cord by Focal Injection of Lysolecithin

Chenyi Wang and Mark R. Kotter

19. Myelin Protein Zero180-199 Peptide Induced Experimental Autoimmune Neuritis in C57BL/6 Mice

David G. Gonsalvez, SangWon Yoo, Georgina A. Craig, Rhiannon J. Wood, Jessica L. Fletcher, Simon S. Murray, and Junhua Xiao

20. Transection and Crush Models of Nerve Injury to Measure Repair and Remyelination in Peripheral Nerve

Xin-peng Dun and David B. Parkinson

21. In Vivo Introduction of Transgenes into Mouse Sciatic Nerve Cells Using Viral Vectors

Gerben Van Hameren, Sergio Gonzalez, Ruani N. Fernando, Claire Perrin-Tricaud, and Nicolas Tricaud

22. Intrathecal Delivery of Viral Vectors for Gene Therapy

Alexia Kagiava and Kleopas A. Kleopa

23. Efficient Gene Transfer to Myelinating Schwann Cells in the Rodent Sciatic Nerve Using In Vivo Electroporation

Daisuke Ino and Masamitsu Iino