Description
This fully updated volume brings together synapse development methods that span from the macroscopic to the nanoscale in order to foster novel perspectives and encourage interdisciplinary integration. Beginning with a section on in vitro formation and manipulation of synapses, including cutting-edge approaches for neuron differentiation and genomic modification, the book continues by covering methods for dissecting synaptic components at high resolution, utilizing advanced imaging and purification techniques, as well as approaches for studying synaptic networks and activity patterns in their native contexts, featuring methods for labeling, tracing, and quantifying components of developing neural circuits. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Synapse Development: Methods and Protocols, Second Edition serves as valuable resource for researchers seeking to advancing our understanding of how synapses form, transform, and transmit.
Table of Contents
Synaptogenic Assays Using Primary Neurons Cultured on Micropatterned Substrates.- Generation of Glutamatergic Human Neurons from Induced Pluripotent Stem Cells.- Generation of Prefrontal Cortex Neurons from Human Pluripotent Stem Cells Under Chemically Defined Conditions.- Differentiation of Mature Dopaminergic Neurons from Human Induced Pluripotent Stem Cells.- Prime Editing of Mouse Primary Neurons.- Purification of Afference-Specific Synaptosome Populations Using Fluorescence-Activated Synaptosome Sorting.- Investigating the Molecular Composition of Neuronal Subcompartments Using Proximity Labeling.- Monitoring Synapses via Trans-Synaptic GFP Complementation.- Imaging Synapse Ultrastructure and Organization with STED Microscopy.- Photomarking Relocalization Technique for Correlated Two-Photon and Electron Microscopy Imaging of Single Stimulated Synapses.- Image Analyses Routines for Quantification of Synaptic Density and Connectivity.- Monosynaptic Tracing in Developing Circuits Using Modified Rabies Virus.- Gene Knockout in the Developing Brain of Wild-Type Rodents by CRISPR In Utero Electroporation.- Mapping Synaptic Activity at the Population and Cellular Levels with Genetically Encoded Voltage Indicators (GEVIs).- Visualization of Synapses in Larval Stages of Drosophila melanogaster Using the GRASP Technique.- Sparse Labeling, Rapid Clearing, and Native Fluorescence Light Sheet Imaging in the Developing Rodent Cerebellum.
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