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
Melissa Miller, Meng Wu, Jia Xu, David Weaver, Min Li, and Michael X. Zhu
Methods to Study TRPC Channel Regulation by Interacting Proteins
Kirill Kiselyov, Kyu Pil Lee, Joseph P. Yuan, and Shmuel Muallem
Activation of TRP Channels in Mammalian Systems
Tomohiro Numata, Shigeki Kiyonaka, Kenta Kato,Nobuaki Takahashi, and Yasuo Mori
Studying Subunit Interaction and Complex Assembly of TRP Channels
Michael Poteser and Klaus Groschner
Proteomic Analysis of TRPC Channels
Timothy Lockwich, Anthony Makusky, Jeffrey A. Kowalak, Sanford P. Markey, and Indu S. Ambudkar
Lessons of Studying TRP Channels with Antibodies
Marcel Meissner, Verena C. Obmann, Michael Hoschke, Sabine Link, Martin Jung, Gerhard Held, Stephan E. Philipp, Richard Zimmermann, and Veit Flockerzi
Assessing TRPC Channel Function Using Pore-Blocking Antibodies
Shang-Zhong Xu
Strategies and Protocols to Generate Mouse Models with Targeted Mutations to Analyze TRP Channel Functions
Marc Freichel, Ulrich Kriebs, Dominik Vogt, Stefanie Mannebach, and Petra Weißgerber
Studying Endogenous TRP Channels in Visceral and Vascular Smooth Muscles
Alexander V. Zholos
TRPC Channels in Neuronal Survival
Junbo Huang, Wanlu Du, Hailan Yao, and Yizheng Wang
Electrophysiological Methods for the Study of TRP Channels
Karel Talavera and Bernd Nilius
High-Throughput Approaches to Study Mechanisms of TRP Channel Activation
Hongzhen Hu, Michael Bandell, Jorg Grandl, and Matt Petrus
Methods Used for Studying TRP Channel Functions in Sensory Neurons
Louis S. Premkumar
Time-Resolved Activation of Thermal TRP Channels by Fast Temperature Jumps
Feng Qin
Fluorescence Microscopy in Thermo TRP Channel Research
Fan Yang and Jie Zheng
Regulation of TRP Channels by Osmomechanical Stress
Min Jin, Jonathan Berrout, and Roger G. O’Neil
Study of TRP Channels in Cancer Cells
V’yacheslav Lehen’kyi and Natalia Prevarskaya
In Vitro and In Vivo Assays for the Discovery of Analgesic
Drugs Targeting TRP Channels
Jun Chen, Regina M. Reilly, Philip R. Kym, and Shailen Joshi
Studying TRP Channels in Intracellular Membranes
Mohammad Samie and Haoxing Xu
Methods for Studying Drosophila TRP Channels
Elkana Kohn and Baruch Minke
Studying TRP Channels in Caenorhabditis elegans
Rui Xiao and X.Z. Shawn Xu
Biography
Michael Xi Zhu is currently with the Department of Integrative Biology and Pharmacology at the University of Texas Health Science Center at Houston