Methods in Signal Transduction Series – serie

This book focuses on neuron signaling in the regulation of metabolism and body weight, and especially on methods used in these studies. Obesity and related metabolic syndromes have reached epidemic status, but still are no effective strategies for prevention and treatment. Body weight homeostasis is maintained by balanced food intake and energy expenditure, both of which are under the control of brain neurons. In the recent years, significant progress has been made in identifying specific neurons, neural pathways, and non-neuron cells in feeding regulation, as well as in delineating autonomic nervous systems targeting peripheral metabolic tissues in the regulation of energy expenditure and metabolism. This book reviews recent progress on important neuron signaling for body weight and metabolic regulation and the state-of-the-art methods that has been applied in this field, ranging from animal models with neuron-specific manipulations, pharmacology, optogenetics, in vivo Ca2+ imaging, and viral tracing. Readers will be exposed to latest research frontiers on neuron regulation of metabolism.Key Features Explores the role signaling between neurons plays with respect to metabolism Documents how neurotransmitters affect the regulation of feeding Describes various methods and technologies used to study the neuronal control of metabolism Includes contributions from an international team of leading researchers.Related TitlesLim, W. & B. Mayer. Cell Signaling: Principles and Mechanisms (ISBN 978-0-8153-4244-1)Feltz, A. Physiology of Neurons (ISBN 978-0-8153-4600-5)Zempleni, J. & K. Dakshinamurti, eds. Nutrients and Cell Signaling (ISBN 978-0367-39307-6)

Inflammation, once considered a physiological response to foreign pathogens, is now recognized as a crucial pathological player in the initiation and progression of several chronic diseases, including diabetes, obesity, cancer, Alzheimer’s disease, Parkinson’s disease, and many others. Considering that cardiovascular diseases are a leading cause of death in the United States and worldwide, the identification of critical inflammatory processes is of utmost importance to devising new immune-based therapeutics that can be added to existing regimens. This book provides detailed information on aspects of inflammation and the manner in which immune activation pathways affect the progression of cardiovascular diseases and the repair/regeneration mechanisms of underlying diseased tissues.Key Features Outlines the role that inflammation plays in cardiovascular diseases Describes a paradox – neutralization of cytokines that contribute to cardiovascular disease does not show benefit Summarizes research on a variety of processes and mechanisms contributing to cardiovascular pathology Contributions from an international team of leading cardiologists and cardiovascular immunologistsRelated TitlesRoy, S., et al., eds. Chronic Inflammation: Molecular Pathophysiology, Nutritional and Therapeutic Interventions (ISBN 978-1-138-19955-2)Kong, A-N. T., ed. Inflammation, Oxidative Stress, and Cancer: Dietary Approaches for Cancer Prevention (ISBN 978-1-138-19984-2)Dick, A., et al. Practical Manual of Intraocular Inflammation (ISBN 978-0-367-38720-4)

Ion channels generate bioelectricity. Recent findings have documented the biophysical properties, the structure, assembly and regulation, and function and dysfunction of nonclassical nervous system ion channels. This book reviews nonclassical ion channel research, ranging from the basic biology, structure, regulations to their functions not only in normal physiology but also neurological disorders, using a variety of cutting-edge techniques and novel animal models.

Ion channels generate bioelectricity. Recent findings have documented the biophysical properties, the structure, assembly and regulation, and function and dysfunction of nonclassical nervous system ion channels. This book reviews nonclassical ion channel research, ranging from the basic biology, structure, regulations to their functions not only in normal physiology but also neurological disorders, using a variety of cutting-edge techniques and novel animal models.

All hollow organs, such as blood vessels, the gastrointestinal tract, airways, male and female reproductive systems, and the urinary bladder are primarily composed of smooth muscle. Such organs regulate flow, propulsion and mixing of luminal contents and storage by the contraction and relaxation of smooth muscle cells. Smooth muscle cells respond to numerous inputs, including pressure, shear stress, intrinsic and extrinsic innervation, hormones and other circulating molecules, as well as autocrine and paracrine factors. This book is a review of smooth muscle cell regulation in the cardiovascular, reproductive, GI, and other organ systems with emphasis on calcium and receptor signaling.Key selling features:Focuses on smooth muscles of different types Describes ion channel signaling mechanismsReviews calcium and receptor signalingIncludes novel, cutting-edge methodologiesSummarizes studies of mice with genetically encoding sensors in smooth muscleChapter 9 of this book is freely available as a downloadable Open Access PDF under a CC-BY 4.0 license. https://s3-us-west-2.amazonaws.com/tandfbis/rt-files/docs/Open+Access+Chapters/9781498774222_oachapter9.pdf

Autophagy and Signaling is an up-to-date overview of the many signaling pathways regulating autophagy in response to different cellular needs. Discussion includes the status and future directions of autophagy signaling research with respect to different aspects of health and disease. These include the roles of autophagy in regulating cell fate, immune response and host defense, nutrient sensing and metabolism, neural functions and homeostasis. The mechanisms and significance of cross-talk between autophagy and other cellular processes is also explored. Lastly, alterations in autophagy observed in aging and age-related pathologies are described.

Calcium Entry Channels in Non-Excitable Cells focuses on methods of investigating the structure and function of non-voltage gated calcium channels. Each chapter presents important discoveries in calcium entry pathways, specifically dealing with the molecular identification of store-operated calcium channels which were reviewed by earlier volumes in the Methods in Signal Transduction series. Crystallographic and pharmacological approaches to the study of calcium channels of epithelial cells are also discussed. Calcium ion is a messenger in most cell types. Whereas voltage gated calcium channels have been studied extensively, the non-voltage gated calcium entry channel genes have only been identified relatively recently. The book will fill this important niche.

T cells play a vital role mediating adaptive immunity, a specific acquired resistance to an infectious agent produced by the introduction of an antigen. There are a variety of T cell types with different functions. They are called T cells, because they are derived from the thymus gland. This volume discusses how T cells are regulated through the operation of signaling mechanisms. Topics covered include positive and negative selection, early events in T cell receptor engagement, and various T cell subsets.

Gap junction channels are a group of intercellular channels expressed in tissues and organs to synchronize many physiological processes. A gap junction channel is formed by the docking of two hemichannels, and each hemichannel is a hexamer of connexins. The field of gap junction channel and hemichannel research has recently exploded and became one of the most active areas of cell biology. Numerous novel approaches and techniques have been developed, but there is no single book dedicated to the unique techniques and protocols employed for the research on these large pore channels. This book fills the gap and focuses on protocols, approaches and reviews of gap junction channels and connexin hemichannels. It will be a useful reference for graduate students, postdoctoral fellows and researchers. Anyone with an interest in gap junction channels and hemichannels will need this summary of state-of-the-art techniques and protocols.

Showcasing the recent progresses of the field, Cyclic Nucleotide Signaling covers the major tools and methodologies used in various areas of research. The majority of the chapters are protocol oriented, with the goal of providing clear directions for laboratory use. Students and investigators new to the field will find this book particularly informative, as will scientists already actively researching nucleotide signaling.

This book focuses on neuron signaling in the regulation of metabolism and body weight, and especially on methods used in these studies. Obesity and related metabolic syndromes have reached epidemic status, but still are no effective strategies for prevention and treatment. Body weight homeostasis is maintained by balanced food intake and energy expenditure, both of which are under the control of brain neurons. In the recent years, significant progress has been made in identifying specific neurons, neural pathways, and non-neuron cells in feeding regulation, as well as in delineating autonomic nervous systems targeting peripheral metabolic tissues in the regulation of energy expenditure and metabolism. This book reviews recent progress on important neuron signaling for body weight and metabolic regulation and the state-of-the-art methods that has been applied in this field, ranging from animal models with neuron-specific manipulations, pharmacology, optogenetics, in vivo Ca2+ imaging, and viral tracing. Readers will be exposed to latest research frontiers on neuron regulation of metabolism.Key Features Explores the role signaling between neurons plays with respect to metabolism Documents how neurotransmitters affect the regulation of feeding Describes various methods and technologies used to study the neuronal control of metabolism Includes contributions from an international team of leading researchers.Related TitlesLim, W. & B. Mayer. Cell Signaling: Principles and Mechanisms (ISBN 978-0-8153-4244-1)Feltz, A. Physiology of Neurons (ISBN 978-0-8153-4600-5)Zempleni, J. & K. Dakshinamurti, eds. Nutrients and Cell Signaling (ISBN 978-0367-39307-6)

Developmental biologists have been driven to investigate growth factor signaling in embryos in order to understand the regulatory mechanisms underlying a given developmental process. Thus, it is critical to explore the technical methods and experimental designs for growth factor signaling in embryos.Focusing on specific pathways or pathway components, Analysis of Growth Factor Signaling in Embryos provides the methods and guidelines for experimental design to study major aspects of cell signaling in vertebrate embryos. The book covers a broad range of topics in signaling and a variety of current model organisms. Section I explores specific signaling pathways or pathway components. In this section, some chapters highlight the biochemistry of signaling pathways during development, which is often distinctive from that observed in cell culture systems. Section II discusses ionic regulatory mechanisms and the two chapters in Section III examine ways of investigating gene regulation in response to extracellular signals. Finally, Section IV addresses emerging strategies that facilitate integrated analyses of cell signaling in vivo in embryonic systems. Featuring contributions from expert researchers, Analysis of Growth Factor Signaling in Embryos will provide a foundation for further explorations of the cellular regulatory mechanisms governing vertebrate embryonic development.

Lipid Second Messengers provides detailed methodology for analysis of various lipid signaling pathways. Authoritative contributors explain the factors that regulate lipid second messenger production by agonist-activated enzymes and examine their products. Topics discussed include procedures used to measure lipid-derived mediators such as lysophospholipids, arachidonic acid, eicosanoids, anandamide, and ceramides, and the enzymes responsible for generating these messengers, such as phospholipases, prostaglandin endoperoxide synthases, and sphingomyelinase.

In the twenty-first century, we are just beginning to understand more clearly the enormous diversity and complexity of signaling processes in the retina. Integrating advances in the biochemistry, cell biology, physiology, and physics of phototransduction, Signal Transduction in the Retina presents the methodologies and experimental approaches that yield key information on the mechanisms underlying normal retinal physiology. This in-depth work discusses the latest techniques and applications for understanding retinal function and degradation, developing novel therapeutic strategies, and promoting cellular survival and functional retention.Drawing contributions from experts in a range of disciplines, each chapter presents a brief overview of the area discussed along with specific methodology for obtaining the primary data to understand the cellular and molecular process. Given the dominance and wealth of information on rhodopsin-based phototransduction, the book devotes substantial attention to this topic, but also evaluates a diversity of signaling mechanisms. Beginning with the molecular mechanisms of vertebrate phototransduction, this volume presents the structure of phototransduction cascade components at atomic resolution, as well as molecular interactions in multi-protein complexes and novel cell-based strategies for understanding signal shut-off and light adaptation. It discusses non-visual phototransduction and the role of melanopsin in adaptive photoresponses and circadian clock regulation. The book also compares the visual signaling processes of vertebrates and invertebrates. It examines experimental studies of insulin-based signaling in the inner and outer retina; investigates retinal development including signaling in progenitor cells, cell-cell communication in developing cells, and neovascularization; and studies lipid-derived mediators such as neuroprotectins and discusses the participation of retinal pigment epithelium in neuronal survival.

As the highly anticipated update to Lipid Second Messengers (CRC Press, 1999), Lipid-Mediating Signaling is a current and comprehensive overview of research methods used in lipid-mediated signal transduction. Pioneering experts provide a much-needed distillation of a decade’s worth of advances in research techniques that are pertinent in understanding how lipid-mediated signal transduction ties to pathologic disorders. Part I describes methods used to identify activities of and assay the expression of different enzymes involved in lipid-mediated signaling. Part II highlights the use of mass spectrometry to ascertain the lipid content in various systems. Part III contains two chapters devoted to techniques used to determine the role of lipids in the activation of gene transcription. Part IV rounds out the book’s solid coverage by focusing on methods used to assess fatty acid uptake and metabolism. Lipid-Mediating Signaling is an unrivaled reference for this rapidly expanding, yet resource-limited field, and it is a valuable resource for for cell biologists, biochemists, and pharmacologists, as well as for researchers studying signaling transduction.

This volume focuses on the investigatory methods applied to autosomal dominant polycystic kidney disease (ADPKD), one of the most common human genetic diseases. ADPKD is caused by mutations in PKD1 and TRPP2, two integral membrane proteins that function as receptor/ion channels in primary cilia of tubular epithelial cells. Thus, ADPKD belongs to ciliopathies, a group of disorders caused by abnormal cilia formation or function. This proposed book will cover the state-of-the-art methods ranging from molecular biology, biochemistry, electrophysiology, to tools in model animal studies.Key FeaturesExplores the role of cilia in polycystic kidney diseaseFocuses on myriad state-of-the-art methods and techniquesReviews specific mutations integral to this autosomal genetic diseaseIncludes discussions of model systems

Lysosomes are key subcellular organelles that regulate the cell function. Many of the essential activities of the cell are dependent on lysosomes. Dysfunction is linked to multiple diseases - storage disorders, neurodegeneration, immunological diseases and cancer. This book discusses concepts and methods used to study lysosome ion and small molecule transport. The contents will not only attract accomplished investigators in need of a broad review and synthesis of this important subject but will also appeal to young investigators and trainees needing to acquire comprehensive knowledge and technical skills working with lysosomal ion channels and small molecule transporters.Key selling features:Summarizes the endocellular role that lysosomes play with respect to cellular waste disposalReviews essential cellular functions of lysosomesExplores how lysosome dysfunction is the cause of many metabolic disordersExamines how lysomes are involved in storage diseasesDescribes various technologies and methods used in lysosome research

New Techniques for Studying Biomembranes describes some of the latest methods used to investigate the dynamic distribution of specific lipids in membranes and their effects on other membrane components. The contributors present important discoveries with respect to lipid analysis and lipid interactions with membrane proteins. Various methods, which have been used to study lipid bilayer structure and lipid organization in membranes, include both in vitro and in vivo membrane systems, and study membrane proteins in various membrane systems.Key Features:Reviews both in vivo and in vitro analytical technologies and methods for studying membrane structure and functionExplores how lipid bilayers and membrane proteins interactIncludes contributions from an international team of researchers actively studying membrane structure and functionIdentifies various diseases whose causes are related to membrane proteinsRelated Titles:Christopher R. Jacobs, Hayden Huang, and Ronald Y. Kwon. Introduction to Cell Mechanics and Mechanobiology (ISBN 978-0-8153-4425-4)Wendell Lim and Bruce Mayer. Cell Signaling: Principles and Mechanisms (ISBN 978-0-8153-4244-1)Stephen Rothman. Proteins Crossing Membranes: A Scientist’s Memoir (978-0-3670-7449-4)

Optogenetics represents a breakthrough technology capable of dynamically modulating molecular and cellular activity in live cells with high precision. This transformative technology made it possible to reversibly interrogate protein actions, cellular events, and animal behaviors with a simple flash of light. Manipulating the temporal and spatial profile of light enabled precise control of membrane potentials. This feature has inspired non-opsin-based optogenetics, which not only inherits the precise spatiotemporal resolution but also expands the targets into a diverse pool of biomolecules such as membrane receptors, ion channels, kinases, GTPases, and transcription factors. This book is unique because it emphasizes the design and applications of opsin-free optogenetic tools. Key Features Describes new developments in non-opsin-based optogenetic tools Introduces cutting-edge methods for precise modulation of cell signaling Highlights applications of optogenetic regulation of cellular functions Covers contributions from an international team of leading expertsRelated Titles Lim, W. & B. Mayer – Cell Signaling (ISBN 9780815342441) Ayyanathan, K., ed. – Cancer Cell Signaling: Targeting Signaling Pathways Toward Therapeutic Approaches to Cancer (ISBN 9781774630839) Zhang , J. et al., eds. – Optical Probes in Biology (ISBN 9781138199934)

Inflammation, once considered a physiological response to foreign pathogens, is now recognized as a crucial pathological player in the initiation and progression of several chronic diseases, including diabetes, obesity, cancer, Alzheimer’s disease, Parkinson’s disease, and many others. Considering that cardiovascular diseases are a leading cause of death in the United States and worldwide, the identification of critical inflammatory processes is of utmost importance to devising new immune-based therapeutics that can be added to existing regimens. This book provides detailed information on aspects of inflammation and the manner in which immune activation pathways affect the progression of cardiovascular diseases and the repair/regeneration mechanisms of underlying diseased tissues.Key Features Outlines the role that inflammation plays in cardiovascular diseases Describes a paradox – neutralization of cytokines that contribute to cardiovascular disease does not show benefit Summarizes research on a variety of processes and mechanisms contributing to cardiovascular pathology Contributions from an international team of leading cardiologists and cardiovascular immunologistsRelated TitlesRoy, S., et al., eds. Chronic Inflammation: Molecular Pathophysiology, Nutritional and Therapeutic Interventions (ISBN 978-1-138-19955-2)Kong, A-N. T., ed. Inflammation, Oxidative Stress, and Cancer: Dietary Approaches for Cancer Prevention (ISBN 978-1-138-19984-2)Dick, A., et al. Practical Manual of Intraocular Inflammation (ISBN 978-0-367-38720-4)

The lungs are the organ for gas exchange between the body and the external environment. Dysfunction of upper airway epithelium and smooth muscle cells leads to pathogenesis of asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis, and other conditions, resulting in airway inflammation and narrowing. Injury to alveolar epithelium and endothelium causes influx of neutrophil and protein-rich fluid from circulation, resulting in edema and disruption of gas exchange. In addition to lung structural cells, immune cells, including alveolar macrophages and lymphocytes play critical roles in the maintenance of lung function. This book contributes to the understanding homeostasis of lung cells in the physiological and pathological conditions critical to the development of novel therapeutics.Key FeaturesHighlights the role the lungs play as an interface between the body and the environmentDescribes the underlying mechanism of lung diseasesEmphasizes the ways nutrition contributes to lung health as well as the ways pollution adversely affect lung functionIncludes contributions from leading researchersChapters 8 and 13 of this book are available for free in PDF format as Open Access from the individual product page at www.routledge.com. It has been made available under a Creative Commons Attribution-Non Commercial-No Derivatives 4.0 license.

The field of transient receptor potential (TRP) channels has gained momentum in recent years not only because of the 2021 Nobel Prize in Physiology or Medicine awarded to David Julius and Ardem Patapoutian for their discoveries of receptors for temperature and touch, but also because of the growing appreciation of the diverse and important physiological and pathophysiological functions of this diverse family of cation channels. In the past decade, there have been important discoveries in the TRP field: resolution of the 3-D structures of major subfamilies using single-particle cryogenic electron microscopy and X-ray crystallography, identification and development of selective agonists and antagonists for most TRP subfamily members, discovery of new human diseases associated with TRP gene mutations, as well as improved understanding of TRP channel function and regulation in various systems. These new discoveries are addressed in TRP-Mediated Signaling, a volume in the Methods in Signal Transduction book series.Key Features• Summarizes recent advances in the study of TRP channels• Written by a team of leading international researchers• Reviews physiological and pathophysiological functions of TRP channels• Identifies diseases associated with TRP gene mutations• Examines the evolution of temperature-sensing TRP channels

As the highly anticipated update to Lipid Second Messengers (CRC Press, 1999), Lipid-Mediating Signaling is a current and comprehensive overview of research methods used in lipid-mediated signal transduction. Pioneering experts provide a much-needed distillation of a decade’s worth of advances in research techniques that are pertinent in understanding how lipid-mediated signal transduction ties to pathologic disorders. Part I describes methods used to identify activities of and assay the expression of different enzymes involved in lipid-mediated signaling. Part II highlights the use of mass spectrometry to ascertain the lipid content in various systems. Part III contains two chapters devoted to techniques used to determine the role of lipids in the activation of gene transcription. Part IV rounds out the book’s solid coverage by focusing on methods used to assess fatty acid uptake and metabolism. Lipid-Mediating Signaling is an unrivaled reference for this rapidly expanding, yet resource-limited field, and it is a valuable resource for for cell biologists, biochemists, and pharmacologists, as well as for researchers studying signaling transduction.

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 channelsPractical protocols for functional studies of TRP channels, including TRPC, TRPV, TRPA, TRPM and the intracellularly localized TRPML channelsThermoTRPs, including the new fast temperature jump apparatus and the high throughput random mutagenesis method for screening critical motifs involved in TRP channel regulationCell-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.

Lipid Second Messengers provides detailed methodology for analysis of various lipid signaling pathways. Authoritative contributors explain the factors that regulate lipid second messenger production by agonist-activated enzymes and examine their products. Topics discussed include procedures used to measure lipid-derived mediators such as lysophospholipids, arachidonic acid, eicosanoids, anandamide, and ceramides, and the enzymes responsible for generating these messengers, such as phospholipases, prostaglandin endoperoxide synthases, and sphingomyelinase.

Lysosomes are key subcellular organelles that regulate the cell function. Many of the essential activities of the cell are dependent on lysosomes. Dysfunction is linked to multiple diseases - storage disorders, neurodegeneration, immunological diseases and cancer. This book discusses concepts and methods used to study lysosome ion and small molecule transport. The contents will not only attract accomplished investigators in need of a broad review and synthesis of this important subject but will also appeal to young investigators and trainees needing to acquire comprehensive knowledge and technical skills working with lysosomal ion channels and small molecule transporters.Key selling features:Summarizes the endocellular role that lysosomes play with respect to cellular waste disposalReviews essential cellular functions of lysosomesExplores how lysosome dysfunction is the cause of many metabolic disordersExamines how lysomes are involved in storage diseasesDescribes various technologies and methods used in lysosome research

This volume focuses on the investigatory methods applied to autosomal dominant polycystic kidney disease (ADPKD), one of the most common human genetic diseases. ADPKD is caused by mutations in PKD1 and TRPP2, two integral membrane proteins that function as receptor/ion channels in primary cilia of tubular epithelial cells. Thus, ADPKD belongs to ciliopathies, a group of disorders caused by abnormal cilia formation or function. This proposed book will cover the state-of-the-art methods ranging from molecular biology, biochemistry, electrophysiology, to tools in model animal studies.Key FeaturesExplores the role of cilia in polycystic kidney diseaseFocuses on myriad state-of-the-art methods and techniquesReviews specific mutations integral to this autosomal genetic diseaseIncludes discussions of model systems

New Techniques for Studying Biomembranes describes some of the latest methods used to investigate the dynamic distribution of specific lipids in membranes and their effects on other membrane components. The contributors present important discoveries with respect to lipid analysis and lipid interactions with membrane proteins. Various methods, which have been used to study lipid bilayer structure and lipid organization in membranes, include both in vitro and in vivo membrane systems, and study membrane proteins in various membrane systems.Key Features: Reviews both in vivo and in vitro analytical technologies and methods for studying membrane structure and function Explores how lipid bilayers and membrane proteins interact Includes contributions from an international team of researchers actively studying membrane structure and function Identifies various diseases whose causes are related to membrane proteinsRelated Titles:Christopher R. Jacobs, Hayden Huang, and Ronald Y. Kwon. Introduction to Cell Mechanics and Mechanobiology (ISBN 978-0-8153-4425-4)Wendell Lim and Bruce Mayer. Cell Signaling: Principles and Mechanisms (ISBN 978-0-8153-4244-1)Stephen Rothman. Proteins Crossing Membranes: A Scientist’s Memoir (978-0-3670-7449-4)

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 channelsPractical protocols for functional studies of TRP channels, including TRPC, TRPV, TRPA, TRPM and the intracellularly localized TRPML channelsThermoTRPs, including the new fast temperature jump apparatus and the high throughput random mutagenesis method for screening critical motifs involved in TRP channel regulationCell-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.