© 2017 • 200 pages, illustrated (28 color and 8 B&W), index
Paperback • $81.90 • ISBN 978-1-621821-21-2

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.

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