Description
Understanding the molecular and cellular mechanisms underlying the development of specific neural circuits is not just an intellectual curiosity but also central to our ability to develop therapeutic approaches to repair damaged pathways in the future. In Neural Development: Methods and Protocols, experts in the field contribute commonly used protocols to facilitate future research in developmental neuroscience. Split into four convenient sections, this detailed volume covers techniques of culturing neurons and glia as well as their growth and differentiation, methods of gene delivery and down regulation, protocols for analyzing axon growth and guidance plus synapse formation, and, finally, basic methods to analyze brain morphology and axon pathways in developing animals. 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. Comprehensive and accessible, Neural Development: Methods and Protocols provides key guidance for students and postdoctoral fellows new to developmental neurobiology. Part I: Neuron and Glial Cell Culture 1. Isolation and Maintenance of Cortical Neural Progenitor Cells In Vitro Mi-Yeon Kim, Byoung-San Moon, and Kang-Yell Choi 2. Neural Induction with a Dopaminergic Phenotype from Human Pluripotent Stem Cells Through a Feeder-Free Floating Aggregation Culture Asuka Morizane, Daisuke Doi, and Jun Takahashi 3. Nucleoside Analog Labeling of Neural Stem Cells and Their Progeny Erik DeBoer and Mladen-Roko Rasin 4. Culture of Dissociated Hippocampal Neurons Yun Peng, Wen-Cheng Xiong, and Lin Mei 5. Culturing Mouse Cerebellar Granule Neurons Tharakeswari Selvakumar and Daniel L. Kilpatrick 6. Preparation of Primary Cultured Dopaminergic Neurons from Mouse Brain Won-Seok Choi, Hyung-Wook Kim, and Zhengui Xia 7. Culturing Astoryctes from Postnatal Rats Yangyang Huang and Cheryl Dreyfus 8. Culturing Oligodendrocyte Lineage Cells from Neonatal Rats Clifton G. Fulmer and Cheryl F. Dreyfus 9. Isolation and Culture of Schwann Cells Yanmei Tao Part II: Gene Expression and Analysis 10. DNA Transfection: Calcium Phosphate Method Munjin Kwon and Bonnie L. Firestein 11. The Gene-Gun Approach for Transfection and Labeling of Cells in Brain Slices Anna Dunaevsky 12. Lentiviral Vector Production, Titration, and Transduction of Primary Neurons Baojin Ding and Daniel L. Kilpatrick 13. In-Ovo Electroporation in Embryonic Chick Spinal Cords Hui Wang and Michael P. Matise 14. Gene Transfer in Developing Chick Embryos: In Ovo Electroporation Emma K. Farley 15. In Utero Electroporation in Mice Chunlei Wang and Lin Mei 16. General Introduction to In Situ Hybridization Protocol Using Non-Radioactively Labeled Probes to Detect mRNAs on Tissue Sections Daehoon Lee, Shan Xiong, and Wen-Cheng Xiong 17. Immunofluorescence Staining with Frozen Mouse or Chick Embryonic Tissue Sections Hui Wang and Michael P. Matise 18. -Galactosidase Staining of LacZ Fusion Proteins in Whole Tissue Preparations Margaret A. Cooper and Renping Zhou 19. Chromatin Immunoprecipitation Assay of Brain Tissues Using Percoll Gradient-Purified Nuclei Baojin Ding and Daniel L. Kilpatrick 20. In Vivo Dual Luciferase Reporter Assay with Chick Neural Tube In Ovo Electroporation System Hui Wang and Michael P. Matise Part III: Axon Pathways and Synapses 21. Growth Cone Collapse Assay Xin Yue, Alexander I. Son, and Renping Zhou 22. The Stripe Assay: Studying Growth Preference and Axon Guidance on Binary Choice Substrates In Vitro Markus Weschenfelder, Franco Weth, Bernd Knll, and Martin Bastmeyer 23. Microcontact Printing of Substrate-Bound Protein Patterns for Cell- and Tissue Culture Martin Fritz and Martin Bastmeyer 24. Semi-Automated Analysis of Dendrite Morphology in Cell Culture Eric S. Sweet, Chris L. Langhammer, Melinda K. Kutzing, and Bonnie L. Firestein 25. Monitoring Synaptic Plasticity by Imaging AMPA Receptor Content and Dynamics on Dendritic Spines Hiroshi Makino and Bo Li 26. Morphological Analysis of Neuromuscular Junctions by Immunofluorescent Staining of Whole-Mount Mouse Diaphragms Haitao Wu and Lin Mei Part IV: Morphological Analyses of the Developing Nervous System 27. Routine Histology Techniques for the Developing and Adult Central Nervous System W. Geoffrey McAuliffe 28. Cryosectioning Alexander I. Son, Katie Sokolowski, and Renping Zhou 29. The Golgi-Cox Method Gitanjali Das, Kenneth Reuhl, and Renping Zhou 30. Neuroanatomical Tract-Tracing Using Biotinylated Dextran Amine Nikolai E. Lazarov 31. Retrograde Tracing Technique for Neonatal Animals Kengo Funakoshi, Akira Yoshikawa, and Yoshitoshi Atobe
