Full 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.
Contents
Isolation and Maintenance of Cortical Neural Progenitor Cells In Vitro.- Neural Induction with a Dopaminergic Phenotype from Human Pluripotent Stem Cells Through a Feeder-Free Floating Aggregation Culture.- Nucleoside Analog Labeling of Neural Stem Cells and Their Progeny.- Culture of Dissociated Hippocampal Neurons.- Culturing Mouse Cerebellar Granule Neurons.- Preparation of Primary Cultured Dopaminergic Neurons from Mouse Brain.- Culturing Astoryctes from Postnatal Rats.- Culturing Oligodendrocyte Lineage Cells from Neonatal Rats.- Isolation and Culture of Schwann Cells.- DNA Transfection: Calcium Phosphate Method.- The Gene-Gun Approach for Transfection and Labeling of Cells in Brain Slices.- Lentiviral Vector Production, Titration, and Transduction of Primary Neurons.- In-Ovo Electroporation in Embryonic Chick Spinal Cords.- Gene Transfer in Developing Chick Embryos: In Ovo Electroporation.- In Utero Electroporation in Mice.- General Introduction to In Situ Hybridization Protocol Using Non-Radioactively Labeled Probes to Detect mRNAs on Tissue Sections.- Immunofluorescence Staining with Frozen Mouse or Chick Embryonic Tissue Sections.- β-Galactosidase Staining of LacZ Fusion Proteins in Whole Tissue Preparations.- Chromatin Immunoprecipitation Assay of Brain Tissues Using Percoll Gradient-Purified Nuclei.- In Vivo Dual Luciferase Reporter Assay with Chick Neural Tube In Ovo Electroporation System.- Growth Cone Collapse Assay.- The Stripe Assay: Studying Growth Preference and Axon Guidance on Binary Choice Substrates In Vitro.- Microcontact Printing of Substrate-Bound Protein Patterns for Cell- and Tissue Culture.- Semi-Automated Analysis of Dendrite Morphology in Cell Culture.- Monitoring Synaptic Plasticity by Imaging AMPA Receptor Content and Dynamics on Dendritic Spines.- MorphologicalAnalysis of Neuromuscular Junctions by Immunofluorescent Staining of Whole-Mount Mouse Diaphragms- Routine Histology Techniques for the Developing and Adult Central Nervous System.- Cryosectioning.- The Golgi-Cox Method.- Neuroanatomical Tract-Tracing Using Biotinylated Dextran Amine.- Retrograde Tracing Technique for Neonatal Animals.
