Series Preface .................................................. v
Preface ....................................................... vii
Contributors ................................................... xv
Part I. Molecular Techniques
1 Production of High-Quality Antibodies for the Study
of Receptors and Ion Channels ................................ 3
Masahiko Watanabe
2 Co-immunoprecipitation from Brain ........................... 19
Xavier Morató, Dasiel O. Borroto-Escuela, Kjell Fuxe,
Victor Fernández-Dueñas, and Francisco Ciruela
3 Subsynaptic Membrane Fractionation .......................... 31
Paula M. Canas and Rodrigo A. Cunha
4 Investigation of Neurotransmitter Receptors in Brain
Slices Using Cell Surface Biotinylation ..................... 39
Elek Molnár
5 Single Nanopardcle Tracking of Surface Ion Channels and
Receptors in Brain Cells .................................... 49
Juan Varela, Julien Dupuis, and Laurent Groc
6 Radioligand Binding Detection of Receptors in Brain
Membranes ................................................... 59
Fuencisla Pilar-Cuéllar, Alvaro Díaz, Emilio Garro-
Martínez, Alicia Martín, Beatriz Romero, and Elsa
M. Valdizán
7 Recombinant Alphavirus-Mediated Expression of Ion Channels
and Receptors in the Brain .................................. 77
Markus U. Ehrengruber and Kenneth Lundstrom
8 Fluorescent Ligands and TR-FRET to Study Receptor-Receptor
Interactions in the Brain ................................... 99
Victor Fernández-Dueñas, Thierry Durroux, and Francisco
Ciruela
9 In Situ Proximity Ligation Assay to Study and Understand
the Distribution and Balance of GPCR Homo- and
Heteroreceptor Complexes in the Brain ...................... 109
Dasiel O. Borroto-Escuela, Beth Hagman, Miles Woolfenden,
Luca Pinton, Antonio Jiménez-Beristain, Julia Oflijan,
Manuel Narvaez, Michael Di Palma, Kristin Feltmann,
Stfano Sartini, Patrizia Ambrogini, Francisco Ciruela,
Riccardo Cuppini, and Kjell Fuxe
Part II. Neuroanatomical Techniques
10 Fluorescent In Situ Hybridization for Sensitive and
Specific Labeling .......................................... 127
Miwako Yamasaki and Masahiko Watanabe
11 Autoradiographic Visualization of G Protein-Coupled
Receptors in Brain ......................................... 143
Rebeca Vidal, Raquel Linge, Maria Josefa Castillo, Josep
Amigó, Elsa M. Valdizán, and Elena Castro
12 Analysis of the Expression Profile and Regional
Distribution of Neurotransmitter Receptors and Ion
Channels in the Central Nervous System Using Histoblots .... 157
Elek Molnár
13 Immunohistochemistry for Ion Channels and Their
Interacting Molecules: Tips for Improving Antibody
Accessibility .............................................. 171
Kohtarou Konno and Masahiko Watanabe
14 Localization of GFP-Tagged Proteins at the Electron
Microscope ................................................. 179
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
15 Pre-embedding Methods for the Localization of Receptors
and Ion Channels ........................................... 191
Rafael Luján
16 Post-embedding Immunohistochemistry in the Localisation
of Receptors and Ion Channels .............................. 211
Rafael Luján and Masahiko Watanabe
17 High-Resolution Localization of Membrane Proteins by
SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL) .... 233
Harumi Harada and Ryuichi Shigemoto
18 Application of Virus Vectors for Anterograde Tract-Tracing
and Single-Neuron Labeling Studies ......................... 247
Hiroyuki Hioki, Hisashi Nakamura, and Takahiro Furuta
19 Analysis of Synaptic Connections at the Electron
Microscopic Level Using Viral Vectors ...................... 267
Takahiro Furuta, Keiko Okamoto-Furuta, and Hiroyuki Hioki
20 Morphological and Neurochemical Characterization of
Electrophysiologically Identified Cells .................... 277
Yoshiyuki Kubota
Part III. Functional Techniques
21 Using Electrophysiology to Study Synaptic and
Extrasynaptic Ionotropic Receptors in Hippocampal Neurons .. 313
Ian D. Coombs and David Soto
22 Biophysical Methods to Analyze Direct G-Protein
Regulation of Neuronal Voltage-Gated Calcium Channels ...... 357
Norbert Weiss and Michel De Waard
23 Electrophysiological Recordings in Behaving Animals ........ 369
Agnès Gruart and José M. Delgado-García
24 Voltammetry in Behaving Animals ............................ 397
Kendra D. Bunner and George V. Rebec
25 In Vivo Brain Microdialysis of Monoamines .................. 415
Jorge E. Ortega, J. Javier Meana, and Luis F. Callado
26 Calcium Transients in Single Dendrites and Spines
of Pyramidal Neurons In Vitro .............................. 435
Rbiannon M. Meredith and Martine R. Groen
27 Dynamic Recording of Membrane Potential from Hippocampal
Neurons by Using a FRET-Based Voltage Biosensor ............ 447
Victor Fernández-Dueñas, Xavier Moratу, Thomas Knbpfel,
and Francisco Ciruela
28 Determination of GPCR-Mediated cAMP Accumulation in Rat
Striatal Synaptosomes ...................................... 455
Jaume Taura, Victor Ferndnáez-Dueñas, and Francisco
Ciruela
29 GPCR-Mediated MAPK/ERK Cascade Activation in Mouse
Striatal Slices ............................................ 465
Maricel Gómez-Soler, Victor Fernández-Dueñas, and
Francisco Ciruela
Subject Index ................................................. 473
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