This second edition volume expands on the previous edition with new information on the latest developments in the field, and the most up-to-date representative methodological information on molecular neuroanatomical and functional techniques that are currently used to study neurotransmitter receptors and ion channels in the brain. The chapters in this book are organized into three parts. Part One discusses molecular immunological and pharmacological aspects related to the detection of receptors and ion channels in the brain. Part Two explores the neuroanatomical techniques devoted to localizing receptors and ion channels within the central nervous system. Part Three looks at functional approaches devoted to ascertaining receptors and ion channels functionality in the brain. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory.
Cutting-edge and co
Table of ContentsSeries Preface…
Preface…
Table of Contents…
Contributing Authors…
Part I Molecular Techniques
1. Production of High-Quality Antibodies for the Study of Receptors and Ion Channels
Masahiko Watanabe
2. Co-Immunoprecipitation from Brain
Xavier Morató, Dasiel O. Borroto-Escuela, Kjell Fuxe, Víctor Fernández-Dueñas, and Francisco Ciruela
3. Sub-Synaptic Membrane Fractionation
Paula M. Canas and Rodrigo A. Cunha
4. Quantitative Analysis of Cell Surface Expressed, Intracellular, and Internalized Neurotransmitter Receptor Populations in Brain Slices Using Biotinylation
Elek Molnár
5. Single Nanoparticle Tracking of Surface Ion Channels and Receptors in Brain Cells
Juan Varela, Julien Dupuis, François Maingret, and Laurent Groc
6. Radioligand Binding Detection of Receptors in Brain Membranes
Fuencisla Pilar-Cuéllar, Emilio Garro-Martínez, Elena Castro, and Álvaro Díaz
7. Recombinant Alphavirus-Mediated Expression of Ion Channels and Receptors in the Brain
Markus U. Ehrengruber and Kenneth Lundstrom
8. Time Resolved-Fluorescence Resonance Energy Transfer using Fluorescent Ligands to Study Native G Protein-Coupled Receptor Heteromerization in Brain
Víctor Fernández-Dueñas, Thierry Durroux, and Francisco Ciruela
9. Study of GPCR Homo and Heteroreceptor Complexes in Specific Neuronal Cell Populations using the In Situ Proximity Ligation Assay
Manuel Narváez, Minerva Crespo-Ramírez, Ramón Fores-Pons, Mariana Pita-Rodríguez, Francisco Ciruela, Malgorzata Filip, Sarah Beggiato, Luca Ferraro, Sergio Tanganelli, Patrizia Ambrogini, Miguel Perezde la Mora, Kjell Fuxe, and Dasiel O. Borroto-Escuela
10. Amplified Luminescent Proximity Homogeneous Assay (Alpha)-Based Technique to Detect GPCR Oligomers in Human Post-Mortem Brain
Marta Valle-León, Víctor Fernández-Dueñas, and Francisco Ciruela
Part II Neuroanatomical Techniques
11. Fluorescent In Situ Hybridization for Sensitive and Specific Labeling
Miwako Yamasaki and Masahiko Watanabe
12. Autoradiographic Visualization of G Protein-Coupled Receptors in Brain
Rebeca Vidal, Fuencisla Pilar-Cuellar, Álvaro Díaz, and Elena Castro
13. Localization of Neurotransmitter Receptor and Ion Channel Proteins in Unfixed Brains using In Situ Immunoblotting
Elek Molnár
14. Immunohistochemistry for Ion Channels and their Interacting Molecules: Tips for Improving Antibody Accessibility
Kohtarou Konno and Masahiko Watanabe
15. Localization of GFP-Tagged Proteins at the Electron Microscope
Sara Gil-Perotin, A. Cebrián-Silla, V. Herranz-Pérez, P. García-Belda, S. Gil-García, M. Fil, J.S. Lee, M.V. Nachury, and José Manuel García-Verdugo
16. Tyramide Signal Amplificación for Immunoelectron Microscopy
María José Ulloa-Navas, Patricia García-Tárraga, Susana González-Granero, Pedro Pérez-Borreda, Vicente Herranz-Pérez, and José Manuel García-Verdugo
17. Pre-Embedding Methods for the Localization of Receptors and Ion Channels
Rafael Luján
18. Post-Embedding Immunohistochemistry in the Localization of Receptors and Ion Channels
Rafael Luján and Masahiko Watanabe
19. High-Resolution Localization and Quantitation of Membrane Proteins by SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL)
Walter A. Kaufmann, David Kleindienst, Harumi Harada, and Ryuichi Shigemoto
20. Pre-Embedding Immunostaining of Brain Tissue and Three-Dimensional Imaging with FIB-SEM
Marta Turégano-López, José Rodrigo-Rodríguez, Lidia Alonso-Nanclares, Juncal González-Soriano, Javier DeFelipe, and Ángel Merchán-Pérez
21. Application of Virus Vectors for Anterograde Tract-Tracing and Single-Neuron Labeling Studies
Hiroyuki Hioki, Hisashi Nakamura, and Takahiro Furuta
22. Efficient Labeling of Neurons and Identification of Postsynaptic Sites Using Adeno-Associated Virus Vector
Megumu Takahashi, Yoko Ishida, Naoya Kataoka, Kazuhiro Nakamura, and Hiroyuki Hioki
23. Analysis of Synaptic Connections at the Electron Microscopic Level using Sindbis Virus Vectors
Takahiro Furuta, Keiko Okamoto-Furuta, and Hiroyuki Hioki
24. Morphological and Neurochemical Characterization of Electrophysiologically Identified Cells
Yoshiyuki Kubota
Part III Functional Techniques
25. Using Electrophysiology to Study Synaptic and Extrasynaptic Ionotropic Receptors in Hippocampal Neurons
Ian D. Coombs and David Soto
26. Biophysical Methods to Analyze Direct G-Protein Regulation of Neuronal Voltage-Gated Calcium Channels
Norbert Weiss and Michel De Waard
27. Electrophysiological Recordings in Behaving Animals
Agnès Gruart and José M. Delgado-García
28. Voltammetry in Behaving Animals
Kendra D. Bunner and George V. Rebec
29. In Vivo Brain Microdialysis of Monoamines
Jorge E. Ortega, Blanca Pérez-Palomar, J. Javier Meana, and Luis F. Callado
30. Optical Control of Brain Receptors using Photoactive Drug in Behaving Animals
Marc López-Cano, Víctor Fernández-Dueñas, and Francisco Ciruela
31. Dynamic Recording of Membrane Potential from Hippocampal Neurons by Using a Fluorescence Resonance Energy Transfer-Based Voltage Biosensor
Víctor Fernández-Dueñas, Xavier Morató, Thomas Knöpfel, and Francisco Ciruela
Chapter 32. Monitoring GPCR-Mediated cAMP Accumulation in Rat Striatal Synaptosomes
Jaume Taura, Víctor Fernández-Dueñas, and Francisco Ciruela
33. GPCR-Mediated MAPK/ERK Cascade Activation in Mouse Striatal Slices
Maricel Gómez-Soler, Víctor Fernández-Dueñas, and Francisco Ciruela
Subject Index List…
