Synapse Development: Methods and Protocols (Methods in Molecular Biology)

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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.

Part I: Forming Synapses in a Dish   1. A Cell Culture System to Investigate the Presynaptic Control of Subsynaptic Membrane Differentiation at the Neuromuscular Junction             Nadine Schmidt, Sreya Basu, Stephan Köger, and Hans Rudolf Brenner   2. Co-Culture Synaptogenic Assay: A New Look at Fluorescence Reporters and Technological Devices             Karen Perez de Arce and Massimiliano Stagi   3. Synaptogenic Assays Using Neurons Cultured on Micropatterned Substrates             Katalin Czöndör and Olivier Thoumine   4. Monitoring Synapses Via Trans-Synaptic GFP Complementation             Theodoros Tsetsenis   5. Generation of Spinal Motor Neurons from Human Pluripotent Stem Cells             David P. Santos and Evangelos Kiskinis   Part II: Analyzing Synaptic Constituents   6. Biochemical Purification of Binding Partners of Synaptic Scaffold Proteins             Bekir Altas, Olaf Jahn, and Hiroshi Kawabe   7. In Situ Protein Binding Assay Using Fc-Fusion Proteins             Nirmala Padmanabhan and Tabrez J. Siddiqui   8. Reconstitution of Synaptic SNAREs into Large Liposomes with Reduced Curvature Stress             Javier M. Hernandez   9. Isolation of Synaptosomes, Synaptic Plasma Membranes, and Synaptic Junctional Complexes             Mary L. Michaelis, Lei Jiang, and Elias K. Michaelis