Description
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. 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 Kger, 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 Czndr 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 10. Purification of Synaptosome Populations Using Fluorescence-Activated Synaptosome Sorting Elisa Luquet, Christoph Biesemann, Annie Munier, and Etienne Herzog Part III: Observing Synaptic Structure and Ultrastructure 11. Optimized Protocol for Imaging Cleared Neural Tissues Using Light Microscopy Yoh Isogai, Douglas Richardson, Catherine Dulac, and Joseph Bergan 12. Structured Illumination Microscopy for the Investigation of Synaptic Structure and Function Soyon Hong, Daniel K. Wilton, Beth Stevens, and Douglas S. Richardson 13. 3D dSTORM Imaging of Fixed Brain Tissue Frank Herrmannsdrfer, Benjamin Flottmann, Siddarth Nanguneri, Varun Venkataramani, Heinz Horstmann, Thomas Kuner, and Mike Heilemann 14. Photomarking Relocalization Technique for Correlated Two-Photon and Electron Microcopy Imaging of Single Stimulated Synapses Miquel Bosch, Jorge Castro, Mriganka Sur, and Yasunori Hayashi 15. 3D Analysis of Synaptic Ultrastructure in Organotypic Hippocampal Slice Culture by High-Pressure Freezing and Electron Tomography Cordelia Imig and Benjamin H. Cooper
