Receptor and Ion Channel Detection in the Brain: Methods and Protocols

Preface

PART I. MOLECULAR TECHNIQUES
1. Production of high-quality antibodies for the study of receptors and ion channels in brain
Masahiko Watanabe (Hokkaido University School of Medicine, Japan)

2. Co-immunoprecipitation from brain
Xavier Morató, Dasiel O. Borroto-Escuela, Kjell Fuxe, Víctor Fernández-Dueñas and Francisco Ciruela (Universitat de Barcelona, Spain)

3. Sub-synaptic membrane fractionation
Paula Canas & Rodrigo Cunha (University of Coimbra, Portugal)

4. Investigation of neurotransmitter receptors in brain slices using cell surface biotinylation
Elek Molnar (University of Bristol, Bristol, UK)

5. Single nanoparticle tracking of surface ion channels and receptors in brain cells.
Juan Varela, Julien Dupuis, and Laurent Groc (University de Bordeaux, France)

6. Radioligand binding detection of receptors in brain membranes
Fuencisla Pilar-Cuéllar, Alvaro Díaz, Emilio Garro-Martínez, Alicia Martín, Beatriz Romero y Elsa M Valdizán (University of Cantabria, Santander, Spain)

7. Recombinant Alphavirus-Mediated Expression of Ion Channels and Receptors in the Brain
Markus U. Ehrengruber & Kenneth Lundstrom (University of Zurich, Switzerland)

8. Fluorescent ligands and TR-FRET to study receptor-receptor interactions in the brain
Víctor Fernández-Dueñas, Thierry Durroux and Francisco Ciruela (Universitat de Barcelona, Spain)

9. In situ Proximity Ligation Assay to study and understand the distribution and balance of GPCR homo- and heteroreceptor complexes in the brain
Dasiel O. Borroto-Escuela, Miles Woolfenden, Luca Pinton, Antonio Jimenez-Beristein, Julia Oflijan, Manuel Narvaez, Michael Di Palma, Kristin Feltmann, Stefano Sartini, Patrizia Ambrogini, Francisco Ciruela, Riccardo Cuppini and Kjell Fuxe (Karolinska Institutet, Sweeden)

PART II. NEUROANATOMICAL TECHNIQUES

10. Fluorescent in situ hybridization for sensitive and specific labeling
Miwako Yamasaki and Masahiko Watanabe (Hokkaido University School of Medicine, Japan)

11. Autoradiographic visualization of G protein-coupled receptors in brain
Rebeca Vidal, Raquel Linge, María Josefa Castillo, Josep Amigó, Elsa M Valdizán, Elena Castro (University of Cantabria, Santander, Spain)

12. Analysis of the expression profile and regional distribution of neurotransmitter receptors and ion channels in the central nervous system using histoblots
Elek Molnar (University of Bristol, UK)

13. Immunohistochemistry for ion channels and their interacting molecules: Tips for improving antibody accessibility
Kohtarou Konno & Masahiko Watanabe (Hokkaido University School of Medicine, Japan)

14. Localization of GFP-tagged proteins at the electron microscope
Gil-Perotin, S, Cebrián-Silla, A, Herranz-Pérez, V, García-Belda, P, Gil-García, S, Fil, M, Lee JS, Nachury MV, García-Verdugo JM (Universidad de Valencia, Valencia, Spain)

15. Pre-embedding methods in the localization of receptors and ion channels
Rafael Luján (Universidad de Castilla-La Mancha, Albacete, Spain)

16. Post-embedding immunohistochemistry in the localization of receptors and ion channels
Rafael Luján & Masahiko Watanabe (UCLM, Albacete, Spain)

17. High-Resolution localization of membrane proteins by SDS-digested freeze-fracture replica labelling (SDS-FRL)
Harumi Harada, Ryuichi Shigemoto (Institute of Science and Technology IST, Austria)

18. Application of virus vectors for anterograde tract-tracing and single-neuron labeling studies
Hiroyuki Hioki, Hisashi Nakamura, Takahiro Furuta (Department of Morphological Brain Science, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan)

19. Analysis of synaptic connections using viral vectors at the electron microscopic level
Takahiro Furata, Keiko Okamoto-Furuta, Hiroyuki Hioki (Department of Morphological Brain Science, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan

20. Morphological and neurochemical characterization of electrophysiologically identified cells
Yoshiyuki Kubota (Division of Cerebral Circuitry, National Institute for Physiological Sciences, 5-1 Myodaiji-Higashiyama, Okazaki, Aichi 444-8787, Japan)

PART III. FUNCTIONAL TECHNIQUES

21. Using electrophysiology to study synaptic and extrasynaptic ionotropic receptors in hippocampal neurons
Ian D. Coombs & David Soto (IDIBELL, Spain)

22. Biophysical methods to analyze direct G-protein regulation of neuronal voltage-gated calcium channels
Norbert Weiss and Michel De Waard (Academy of Sciences of the Czech Republic, Czech Republic)

23. Electrophysiological recordings in behaving animals
Agnès Gruart and José M. Delgado-García (UPO, Spain)

24. Voltammetry in behaving animals
Kendra D. Bunner and George V. Rebec (Indiana University, USA)

25. In vivo brain microdialysis of monoamines
Jorge E. Ortega, J. Javier Meana, Luis F. Callado (UPV, Spain)

26. Calcium transients in single dendrites and spines of pyramidal neurons in vitro
Rhiannon M. Meredith and Martine R. Groen (VU University, The Netherlands)

27. Dynamic recording of membrane potential from hippocampal neurons by using a FRET-based voltage biosensor
Víctor Fernández-Dueñas, Xavier Morató, Thomas Knöpfel and Francisco Ciruela
(Universitat de Barcelona, Spain)

28. Determination of GPCR-mediated cAMP accumulation in rat striatal synaptosomes
Jaume Taura, Víctor Fernández-Dueñas & Francisco Ciruela (Universitat de Barcelona, Spain)

29. GPCR-mediated MAPK/ERK cascade activation in brain slices.
Maricel Gómez-Soler, Víctor Fernández-Dueñas & Francisco Ciruela (Universitat de Barcelona, Spain)

Index