Table Of ContentK10983_cover_revised.fhmx 3/23/11 9:40 AM Page 1
C M Y CM MY CY CMY K
BIOLOGICAL SCIENCES
TRP CHANNELS TRP CHANNELS
T
The rapid expansion of the TRP field has generated a large amount of excellent original work
R
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 P
and genetic diversity, as well as the different physiological roles they play, techniques used
C
for studying TRP channels range from single molecular analysis to behavioral animal studies.
Methods in multiple areas, such as molecular biology, fluorescence imaging, electrophysiology, H
cell biology, genetics, proteomics, pharmacology, system physiology, and behavioral assessment, A
are employed to investigate various aspects of these channels. Choosing among many possible
N
topics in these broad areas was a daunting task.
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A comprehensive review of the field, TRP Channels spans the information gap by providing E
broad coverage of current methods and techniques commonly used in TRP channel research, L
and detailed protocols with thorough discussions of the advantages and disadvantages across S
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 Z
h
variety of channel types that serve different physiological functions, highlighting distinctive u
views on how the methodology can be utilized. Some chapters include discussion on the Michael X. Zhu
Edited by
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.
K10983
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an informa business 2N7e0w M Yoardki,s oNnY A 1v0e0n1u6e ETHODS METHODS IN SIGNAL TRANSDUCTION SERIES
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TRP CHANNELS
METHODS IN SIGNAL TRANSDUCTION SERIES
Joseph Eichberg, Jr., Series Editor
Published Titles
Lipid Second Messengers, Suzanne G. Laychock and Ronald P. Rubin
G Proteins: Techniques of Analysis, David R. Manning
Signaling Through Cell Adhesion Molecules, Jun-Lin Guan
G Protein-Coupled Receptors, Tatsuya Haga and Gabriel Berstein
Calcium Signaling, James W. Putney, Jr.
G Protein-Coupled Receptors: Structure, Function, and Ligand Screening,
Tatsuya Haga and Shigeki Takeda
Calcium Signaling, Second Edition, James W. Putney, Jr.
Analysis of Growth Factor Signaling in Embryos, Malcolm Whitman and
Amy K. Sater
Signal Transduction in the Retina, Steven J. Fliesler and Oleg G. Kisselev
Signaling by Toll-Like Receptors, Gregory W. Konat
Lipid-Mediated Signaling, Eric J. Murphy and Thad A. Rosenberger
TRP Channels, Michael X. Zhu
TRP CHANNELS
Edited by
Michael X. Zhu
The University of Texas Health Science Center
Boca Raton London New York
CRC Press is an imprint of the
Taylor & Francis Group, an informa business
CRC Press
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Version Date: 2011902
International Standard Book Number-13: 978-1-4398-1861-9 (eBook - PDF)
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Contents
Series Preface ............................................................................................................ix
Preface.......................................................................................................................xi
Editor .......................................................................................................................xv
Contributors ...........................................................................................................xvii
Chapter 1 High-Throughput Screening of TRPC Channel Ligands Using
Cell-Based Assays ................................................................................1
Melissa Miller, Meng Wu, Jia Xu, David Weaver, Min Li,
and Michael X. Zhu
Chapter 2 Methods to Study TRPC Channel Regulation by
Interacting Proteins ............................................................................21
Kirill Kiselyov, Kyu Pil Lee, Joseph P. Yuan, and Shmuel Muallem
Chapter 3 Activation of TRP Channels in Mammalian Systems .......................43
Tomohiro Numata, Shigeki Kiyonaka, Kenta Kato,
Nobuaki Takahashi, and Yasuo Mori
Chapter 4 Studying Subunit Interaction and Complex Assembly of
TRP Channels ....................................................................................91
Michael Poteser and Klaus Groschner
Chapter 5 Proteomic Analysis of TRPC Channels ...........................................113
Timothy Lockwich, Anthony Makusky, Jeffrey A. Kowalak,
Sanford P. Markey, and Indu S. Ambudkar
Chapter 6 Lessons of Studying TRP Channels with Antibodies ......................135
Marcel Meissner, Verena C. Obmann, Michael Hoschke,
Sabine Link, Martin Jung, Gerhard Held, Stephan E. Philipp,
Richard Zimmermann, and Veit Flockerzi
Chapter 7 Assessing TRPC Channel Function Using Pore-
Blocking Antibodies .........................................................................149
Shang-Zhong Xu
v
vi Contents
Chapter 8 Strategies and Protocols to Generate Mouse Models with
Targeted Mutations to Analyze TRP Channel Functions ................167
Marc Freichel, Ulrich Kriebs, Dominik Vogt,
Stefanie Mannebach, and Petra Weißgerber
Chapter 9 Studying Endogenous TRP Channels in Visceral and Vascular
Smooth Muscles ...............................................................................195
Alexander V. Zholos
Chapter 10 TRPC Channels in Neuronal Survival .............................................219
Junbo Huang, Wanlu Du, Hailan Yao, and Yizheng Wang
Chapter 11 Electrophysiological Methods for the Study of TRP Channels .......237
Karel Talavera and Bernd Nilius
Chapter 12 High-Throughput Approaches to Studying Mechanisms
of TRP Channel Activation ..............................................................255
Hongzhen Hu, Michael Bandell, Jorg Grandl, and Matt Petrus
Chapter 13 Methods Used for Studying TRP Channel Functions
in Sensory Neurons ..........................................................................273
Louis S. Premkumar
Chapter 14 Time-Resolved Activation of Thermal TRP Channels by Fast
Temperature Jumps ..........................................................................299
Feng Qin
Chapter 15 Fluorescence Microscopy in ThermoTRP Channel Research .........319
Fan Yang and Jie Zheng
Chapter 16 Regulation of TRP Channels by Osmomechanical Stress ...............353
Min Jin, Jonathan Berrout, and Roger G. O’Neil
Chapter 17 Study of TRP Channels in Cancer Cells ..........................................375
V’yacheslav Lehen’kyi and Natalia Prevarskaya
Contents vii
Chapter 18 In Vitro and in Vivo Assays for the Discovery of Analgesic
Drugs Targeting TRP Channels .......................................................395
Jun Chen, Regina M. Reilly, Philip R. Kym, and Shailen Joshi
Chapter 19 Studying TRP Channels in Intracellular Membranes ......................411
Mohammad Samie and Haoxing Xu
Chapter 20 Methods for Studying Drosophila TRP Channels ...........................423
Elkana Kohn and Baruch Minke
Chapter 21 Studying TRP Channels in Caenorhabditis elegans .......................447
Rui Xiao and X. Z. Shawn Xu
