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Ion Channels: A Laboratory Manual


Subject Area(s):  BiochemistryBiophysicsNeurobiology

Edited by Paul J. Kammermeier, University of Rochester Medical Center; Ian Duguid, Centre for Integrative Physiology, School of Biomedical Sciences, Edinburgh; Stephan Brenowitz, Howard Hughes Medical Institute

Download a Free Excerpt from Ion Channels: A Laboratory Manual:

Index
Voltage-Gated Na+ Channels: Not Just for Conduction
Sample Protocol: Combining Optogenetics and Electrophysiology to Analyze Projection Neuron Circuits


© 2017 • 200 pages, illustrated (28 color and 8 B&W), index
Hardcover • £94 75.20 (click here to price in US Dollars)
ISBN  978-1-621821-20-5
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Description

The propagation of information through the nervous system depends on rapid changes in the electrical potential across excitable cell membranes and the generation of action potentials—the fundamental unit of neuronal communication. Changes in membrane potential occur when ion channels—pore-forming membrane proteins—open to facilitate the passage of specific ions (e.g., Na+, K+) across the cell membrane in response to a stimulus. Defects in ion channels can lead to a wide range of diseases of the nervous system.

This laboratory manual provides state-of-the-art techniques for investigating ion channel properties and function, particularly in the nervous system. Contributors present electrophysiological methods to examine single-channel activity in cultured cells, to study synaptic plasticity and circuit dynamics in brain slice preparations, and to perform whole-cell recordings in awake and freely moving animals. The use of optogenetic tools to study cellular and circuit dynamics in organotypic slice cultures is also covered. Many of the protocols can be adapted for other ion channels, cell types, and experimental preparations.

The manual includes background on the structure, function, and regulation of different voltage- and ligand-gated ion channels. Therefore, it is a useful resource for all cell biologists and neuroscientists seeking to further understand the complex roles of ion channels in physiology and disease.

Contents

CHAPTER 1 Ion Channels: History, Diversity, and Impact
INTRODUCTION
Ion Channels: History, Diversity, and Impact
Stephan Brenowitz, Ian Duguid, and Paul J. Kammermeier

CHAPTER 2 Voltage-Gated Potassium Channels: A Structural Examination of Selectivity and Gating
INTRODUCTION
Voltage-Gated Potassium Channels: A Structural Examination of Selectivity and Gating
Dorothy M. Kim and Crina M. Nimigean

CHAPTER 3 Voltage-Gated Na+ Channels: Not Just for Conduction
INTRODUCTION
Voltage-Gated Na+ Channels: Not Just for Conduction
Larisa C. Kruger and Lori L. Isom

CHAPTER 4 Measuring Ca2+-Dependent Modulation of Voltage-Gated Ca2+ Channels in HEK-293T Cells
PROTOCOL
1 Measuring Ca2+-Dependent Modulation of Voltage-Gated Ca2+ Channels in HEK-293T Cells
Jessica R. Thomas and Amy Lee

CHAPTER 5 Strategies for Investigating G-Protein Modulation of Voltage-Gated Ca2+ Channels
INTRODUCTION
Strategies for Investigating G-Protein Modulation of Voltage-Gated Ca2+ Channels
Van B. Lu and Stephen R. Ikeda

PROTOCOL
1 G-Protein Modulation of Voltage-Gated Ca2+ Channels from Isolated Adult Rat Superior Cervical Ganglion Neurons
Van B. Lu and Stephen R. Ikeda

CHAPTER 6 Hyperpolarization-Activated Cyclic Nucleotide-Gated Channel Currents in Neurons
INTRODUCTION
Hyperpolarization-Activated Cyclic Nucleotide-Gated Channel Currents in Neurons
Mala M. Shah

PROTOCOL
1 Recording Hyperpolarization-Activated Cyclic Nucleotide-Gated Channel Currents (/h) in Neurons
Mala M. Shah

CHAPTER 7 Single-Channel Recording of Ligand-Gated Ion Channels
INTRODUCTION
Single-Channel Recording of Ligand-Gated Ion Channels
Andrew J.R. Plested

PROTOCOL
1 Single-Channel Recording of Glycine Receptors in Human Embryonic Kidney (HEK) Cells
Andrew J.R. Plested and Jelena Baranovic

CHAPTER 8 Measuring the Basic Physiological Properties of Synapses
INTRODUCTION
Measuring the Basic Physiological Properties of Synapses
Matthew A. Xu-Friedman

PROTOCOL
1 Preparing Brain Slices to Study Basic Synaptic Properties
Matthew A. Xu-Friedman

CHAPTER 9 Cellular and Synaptic Properties of Local Inhibitory Circuits
INTRODUCTION
Cellular and Synaptic Properties of Local Inhibitory Circuits
Court Hull

PROTOCOL
1 Measuring Feedforward Inhibition and Its Impact on Local Circuit Function
Court Hull

CHAPTER 10 Fast Cholinergic Synaptic Transmission in the Mammalian Central Nervous System
INTRODUCTION
Fast Cholinergic Synaptic Transmission in the Mammalian Central Nervous System
Michael Beierlein

PROTOCOL
1 Examining Cholinergic Synaptic Signaling in the Thalamic Reticular Nucleus
Rajan Dasgupta, Frederik Seibt, Yan-Gang Sun, and Michael Beierlein

CHAPTER 11 In Vitro Investigation of Synaptic Plasticity
INTRODUCTION
In Vitro Investigation of Synaptic Plasticity
Therese Abrahamsson, Txomin Lalanne, Alanna J. Watt, and P. Jesper Sjöström

PROTOCOLS
1 Long-Term Potentiation by Theta-Burst Stimulation Using Extracellular Field Potential Recordings in Acute Hippocampal Slices
Therese Abrahamsson, Txomin Lalanne, Alanna J. Watt, and P. Jesper Sjöström

2 Using Multiple Whole-Cell Recordings to Study Spike-Timing-Dependent Plasticity in Acute Neocortical Slices
Txomin Lalanne, Therese Abrahamsson, and P. Jesper Sjöström

CHAPTER 12 Stimulating Neurons with Heterologously Expressed Light-Gated Ion Channels
INTRODUCTION
Stimulating Neurons with Heterologously Expressed Light-Gated Ion Channels
J. Simon Wiegert, Christine E. Gee, and Thomas G. Oertner

PROTOCOLS
1 Preparation of Slice Cultures from Rodent Hippocampus
Christine E. Gee, Iris Ohmert, J. Simon Wiegert, and Thomas G. Oertner

2 Viral Vector-Based Transduction of Slice Cultures
J. Simon Wiegert, Christine E. Gee, and Thomas G. Oertner

3 Single-Cell Electroporation of Neurons
J. Simon Wiegert, Christine E. Gee, and Thomas G. Oertner

CHAPTER 13 Combining Optogenetics and Electrophysiology to Analyze Projection Neuron Circuits
PROTOCOL
1 Combining Optogenetics and Electrophysiology to Analyze Projection Neuron Circuits
Naoki Yamawaki, Benjamin A. Suter, Ian R. Wickersham, and Gordon M.G. Shepherd

CHAPTER 14 Whole-Cell Recording in the Awake Brain
INTRODUCTION
Whole-Cell Recording in the Awake Brain
Doyun Lee and Albert K. Lee

PROTOCOLS
1 In Vivo Patch-Clamp Recording in Awake Head-Fixed Rodents
Doyun Lee and Albert K. Lee

2 Efficient Method for Whole-Cell Recording in Freely Moving Rodents Using Ultraviolet-Cured Collar-Based Pipette Stabilization
Doyun Lee and Albert K. Lee

APPENDIX General Safety and Hazardous Material Information
INDEX