TRP Channels: Methods and Protocols

Description

This book aims to provide a guide to researchers on how to work with Transient Receptor Potential (TRP) channels, describing current methodologies and protocols, and highlighting the challenges ahead that will require the development of new technologies. Articles cover a wide range of techniques, from structure to function, concluding with how to best to evaluate some of the in vivo function of TRP channels, particularly in regard to their involvement in hyperalgesia and allodynia. Also, the collection describes important and inspiring available tools that are currently being used to investigate the activity of these channels in vitro and in vivo in terms of their underlying protein structure, segueing into dealing with the generation of pharmacological tools to explore their in vivo activity and their involvement in cellular signaling pathways, which may be developed into drugs to treat diseases caused by channel dysfunction or hyperactivity. Written for the highly successful Methods in Molecular Biology series, chapters include the necessary detail and implementation advice to ensure successful results in the lab. Authoritative and practical, TRP Channels: Methods and Protocols is a useful set of methodologies for scientists seeking to better understand this vital family of channels. Front Matter ....Pages i-x Molecular Evolution Bioinformatics Toward Structural Biology of TRPV1-4 Channels (Pau Doñate-Macián, Alba Crespi-Boixader, Alex Perálvarez-Marín)....Pages 1-21 Expression, Purification, and Crystallization of the Transient Receptor Potential Channel TRPV6 (Appu K. Singh, Luke L. McGoldrick, Alexander I. Sobolevsky)....Pages 23-37 Cryo Electron Microscopy of TRP Channels (Amrita Samanta, Taylor E. T. Hughes, Vera Y. Moiseenkova-Bell)....Pages 39-50 Mass Spectrometry-Based Analysis of TRPP2 Phosphorylation (Verónica I. Dumit, Michael Köttgen, Alexis Hofherr)....Pages 51-64 Combining Structural Data with Computational Methodologies to Investigate Structure–Function Relationships in TRP Channels (Victoria Oakes, Carmen Domene)....Pages 65-82 Characterization of TRPC Channels in a Heterologous System Using Calcium Imaging and the Patch-Clamp Technique (Elvira de la Peña, Ana Gomis)....Pages 83-97 Fluorescence-Based Functional Assays for Ca2+-Permeable ThermoTRP Channels (Celia Cordero-Sánchez, Irene Mudarra-Fraguas, Asia Fernández-Carvajal)....Pages 99-110 High-Content Imaging of Immunofluorescently Labeled TRPV1-Positive Sensory Neurons (Jörg Isensee, Tim Hucho)....Pages 111-124 Patch-Clamp Combined with Fast Temperature Jumps to Study Thermal TRP Channels (Beiying Liu, Feng Qin)....Pages 125-141 TRP Channel Reconstitution in Lipid Bilayers (Eleonora Zakharian)....Pages 143-166 Methods for Investigating TRP Channel Gating (Osvaldo Alvarez, Karen Castillo, Emerson Carmona, Carlos Gonzalez, Ramon Latorre)....Pages 167-185 In Silico Approaches for TRP Channel Modulation (Magdalena Nikolaeva Koleva, Gregorio Fernandez-Ballester)....Pages 187-206 Drug Discovery for Soft Drugs on TRPV1 and TRPM8 Channels Using the Passerini Reaction (Tracey Pirali, Ubaldina Galli, Marta Serafini, Alessia Griglio, Armando A. Genazzani, Gian Cesare Tron)....Pages 207-221 In Vivo Methods to Study ThermoTRP Channels in Rodents (Sara González-Rodríguez)....Pages 223-231 Back Matter ....Pages 233-238