Synapse Development

This detailed volume collects protocols for experimentation into how neurons connect to produce the extraordinary functionalities of the nervous system. Contributed by experts and pioneers in their respective techniques, the book covers synapses in the brain and in culture, their constituents, their structures, their dynamics, and the assemblies they form, all in the structure of a laboratory guide. Written for the highly successful Methods in Molecular Biology series, chapters include brief 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.
Authoritative and practical, Synapse Development: Methods and Protocols serves as an ideal guide to minimizing the barrier to entry for the integration of new approaches with existing expertise, producing syntheses that will foster novel perspectives on the many ways in which synapses form, transform, and transmit.

Includes cutting-edge methods and protocols for the study of synapse development Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

A Cell Culture System to Investigate the Presynaptic Control of Subsynaptic Membrane Differentiation at the Neuromuscular Junction.- Co-Culture Synaptogenic Assay: A New Look at Fluorescence Reporters and Technological Devices.- Synaptogenic Assays Using Neurons Cultured on Micropatterned Substrates.- Monitoring Synapses Via Trans-Synaptic GFP Complementation.- Generation of Spinal Motor Neurons from Human Pluripotent Stem Cells.- Biochemical Purification of Binding Partners of Synaptic Scaffold Proteins.- In Situ *Protein Binding Assay Using Fc-Fusion Proteins.- ation Microscopy for the Investigation of Synaptic Structure and Function.- 3D d*STORM Imaging of Fixed Brain Tissue.- Photomarking Relocalization Technique for Correlated Two-Photon and Electron Microcopy Imaging of Single Stimulated Synapses.- 3D Analysis of Synaptic Ultrastructure in Organotypic Hippocampal Slice Culture by High-Pressure Freezing and ElectronTomography.- Analyzing Endosomal Docking, Fusion, Sorting, and Budding Mechanisms in Isolated Organelles.- Concurrent Imaging of Receptor Trafficking and Calcium Dynamics by Spinning Disk Confocal Microscopy.- Imaging Activity-Dependent Signaling Dynamics at the Neuronal Synapse Using FRET-Based Biosensors.- Analyzing Structural Plasticity of Dendritic Spines in Organotypic Slice Culture.- Using Fluorescent Markers to Estimate Synaptic Connectivity In Situ.- Dual Anterograde and Retrograde Viral Tracing of Reciprocal Connectivity.- Mapping Synaptic Inputs of Developing Neurons Using Calcium Imaging.- Monosynaptic Tracing in Developing Circuits Using Modified Rabies Virus.