TRP Channels Methods in Signal Transduction Series

The rapid expansion of the TRP field has generated a large amount of excellent original work across many different research fields. However, investigators are not necessarily familiar with the pros and cons of the variety of methods used to study TRP channels. Because of functional and genetic diversity, as well as the different physiological roles they play, techniques used for studying TRP channels range from single molecular analysis to behavioral animal studies. Methods in multiple areas, such as molecular biology, fluorescence imaging, electrophysiology, cell biology, genetics, proteomics, pharmacology, system physiology, and behavioral assessment, are employed to investigate various aspects of these channels. Choosing among many possible topics in these broad areas was a daunting task.

A comprehensive review of the field, TRP Channels spans the information gap by providing broad coverage of current methods and techniques commonly used in TRP channel research, and detailed protocols with thorough discussions of the advantages and disadvantages across methods. Some topics covered include

• Mammalian, Drosophila and C. elegans TRP channels
• Practical protocols for functional studies of TRP channels, including TRPC, TRPV, TRPA, TRPM and the intracellularly localized TRPML channels
• ThermoTRPs, including the new fast temperature jump apparatus and the high throughput random mutagenesis method for screening critical motifs involved in TRP channel regulation
• Cell-based high-throughput screening assays for TRP channels and their applications in drug discoveries
• TRP channel functions in native cells, including smooth muscles, neurons, and cancers

Showcasing the current status of the field, TRP Channels covers the major techniques used in various areas of research. The majority of the chapters are protocol oriented, with the goal of providing clear directions for laboratory use. Because of the breadth of the TRP field, the applications of some methods are described in multiple chapters by experts working on a variety of channel types that serve different physiological functions, highlighting distinctive views on how the methodology can be utilized. Some chapters include discussion on the usefulness and pitfalls associated with the use of certain techniques. Together with chapters that offer comprehensive reviews on the functional regulation and other roles of TRP channels, students and investigators new to the field should find this book particularly informative.

High-Throughput Screening of TRPC Channel Ligands Using Cell-Based Assays. Methods to Study TRPC Channel Regulation by Interacting Proteins. Activation of TRP Channels in Mammalian Systems. Studying Subunit Interaction and Complex Assembly of TRP Channels. Proteomic Analysis of TRPC Channels. Lessons of Studying TRP Channels with Antibodies. Assessing TRPC Channel Function Using Pore-Blocking Antibodies. Strategies and Protocols to Generate Mouse Models with Targeted Mutations to Analyze TRP Channel Functions. Studying Endogenous TRP Channels in Visceral and Vascular Smooth Muscles. TRPC Channels in Neuronal Survival. Electrophysiological Methods for the Study of TRP Channels. High-Throughput Approaches to Study Mechanisms of TRP Channel Activation. Methods Used for Studying TRP Channel Functions in Sensory Neurons. Time-Resolved Activation of Thermal TRP Channels by Fast Temperature Jumps Fluorescence Microscopy in Thermo TRP Channel Research. Regulation of TRP Channels by Osmomechanical Stress. Study of TRP Channels in Cancer Cells. In Vitro and In Vivo Assays for the Discovery of Analgesic Drugs Targeting TRP Channels. Studying TRP Channels in Intracellular Membranes. Methods for Studying Drosophila TRP Channels. Studying TRP Channels in Caenorhabditis elegans. Index.

Michael Xi Zhu is currently with the Department of Integrative Biology and Pharmacology at the University of Texas Health Science Center at Houston