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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; Stephen Brenowitz, NIDCD / National Institutes of Health

Due April 2017 • 500 pages (approx.), illustrated (color and B&W), index
Hardcover • $150 120.00 (click here to price in UK Pounds)
ISBN  978-1-621821-20-5
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Description

The propagation of signals through the nervous system depends on rapid changes in electric potential across cell membranes. These changes are mediated by ion channels—macromolecular pores that facilitate the passage of specific ions (e.g., K+ or Na+) through cell membranes in response to various signals. Defects in ion channels can lead to diseases such as epilepsy.

This laboratory manual provides state-of-the-art techniques for investigating ion channel properties and activity, 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 even freely moving—animals. The use of optogenetic tools to study synapses or small networks in organotypic slice cultures is also covered. Many of the experimental setups described can be adapted for other ion channels, cell types, or systems.

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 normal physiology and disease.

Contents

(preliminary)
Chapter 1
INTRODUCTION
Overview
Paul J. Kammermeier, Ian Duguid, and Stephan Brenowitz

Chapter 2
PROTOCOL
Measuring Ca2+-Dependent Modulation of  Voltage-Gated Ca2+ Channels in HEK-293T cells
Jessica R. Thomas and Amy Lee

Chapter 3
INTRODUCTION
Modulation of Voltage-Gated Ca2+ Channels by G Proteins
Van B. Lu and Stephen R. Ikeda

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

Chapter 4
INTRODUCTION
Single-Channel Recording of Ligand-Gated Ion Channels
Andrew J.R. Plested

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

Chapter 5
INTRODUCTION
Measuring the Basic Physiological Properties of Synapses
Matthew A. Xu-Friedman

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

Chapter 6
INTRODUCTION
Fast Cholinergic Synaptic Transmission in the Mammalian CNS
Michael Beierlein

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

Chapter 7
INTRODUCTION
In Vitro Investigation of Synaptic Plasticity
Therese Abrahamsson, Txomin Lalanne, Alanna J. Watt, and Per Jesper Sjöström

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

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

Chapter 8
INTRODUCTION
Voltage-Gated Potassium Channels: A Structural Examination of  Selectivity and Gating
Dorothy M. Kim and Crina M. Nimigean

Chapter 9
INTRODUCTION
Voltage-Gated Na+ Channels: Not Just for Conduction
Larisa Kruger and Lori L. Isom

Chapter 10
INTRODUCTION
Cellular and Synaptic Properties of Local Inhibitory Circuits
Court Hull

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

Chapter 11
INTRODUCTION
Whole-Cell Recording in the Awake Brain
Doyun Lee and Albert K. Lee

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

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

Chapter 12
INTRODUCTION
Stimulating Neurons with Light-Gated Channels
J. Simon Wiegert, Christine Gee, and Thomas Oertner

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

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

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

Chapter 13
INTRODUCTION
Hyperpolarization-Activated Cyclic Nucleotide–Gated (HCN) Channel  Currents in Neurons
Mala M. Shah

PROTOCOL
Recording Hyperpolarization-Activated Cyclic Nucleotide–Gated (HCN)  Channel Currents (Ih) in Neurons
Mala M. Shah

Chapter 14
PROTOCOL
Combining Optogenetics and Electrophysiology to Analyze Projection Neuron Circuits
Naoki Yamawaki, Benjamin A. Suter, Ian R. Wickersham, and Gordon M. G. Shepherd

APPENDIX General Safety and Hazardous Material Information
Index