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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.