Electrophysiology: Basics, Methods, Modern Approaches and Applications

Preface
Acknowledgements
About This Book
Important Physical Units
Contents
About the Authors
Abbreviations
Chapter 1: Introduction
	1.1 Basic Background Knowledge
	1.2 History of Electrophysiology
		Take-Home Messages
	References
Chapter 2: Basics Theory
	2.1 Electrical Characteristics of Biological Membranes
	2.2 Ion Distribution at Biological Membranes
	2.3 Donnan Distribution and Nernst Equation
		2.3.1 Donnan Distribution
		2.3.2 Nernst Equation
	2.4 Goldman-Hodgkin-Katz Equation
		Take-Home Messages
	References
Chapter 3: Basics: Methods
	3.1 Recording Electrical Signals from Body Surface
	3.2 The Example (ECG)
		3.2.1 Electrophysiological Basics
		3.2.2 Activation of the Heart Muscle
	3.3 Recording Electrical Signals from Tissue
		3.3.1 Intracardiac Electrograms
		3.3.2 The Ussing Chamber
		3.3.3 Recording from the Brain
		3.3.4 Recording Extracellular Field Potentials with Multielectrode Arrays
	3.4 Recording Electrical Signals from Single Cells
		3.4.1 The Ag/AgCl Electrode
		3.4.2 The Microelectrode
		3.4.3 Ion-Selective Microelectrodes
			3.4.3.1 Construction of Ion-Selective Microelectrodes
			3.4.3.2 Theory of Ion-Selective Microelectrodes
		3.4.4 The Carbon-Fibre Technique
			3.4.4.1 Construction of Carbon-Fibre Microelectrodes
			3.4.4.2 Theory of Carbon-Fibre Microelectrodes
			3.4.4.3 Amperometric and Cyclic Voltammetric Measurements
		3.4.5 Basics of Voltage Clamp
			3.4.5.1 The Ideal Voltage Clamp
			3.4.5.2 The Real Voltage Clamp
			3.4.5.3 The Voltage Clamp with Two Electrodes
			3.4.5.4 One-Electrode Voltage Clamp Used for the Patch-Clamp Technique
			3.4.5.5 Performing Voltage Clamp
		3.4.6 Noise in Electrophysiological Measurements
			3.4.6.1 Thermal Noise
			3.4.6.2 Shot Noise
			3.4.6.3 Dielectric Noise
			3.4.6.4 Digitisation Noise
			3.4.6.5 The Sampling Theorem and Aliasing Noise
			3.4.6.6 Excess Noise
				Take-Home Messages
	References
Chapter 4: Application of the Voltage-Clamp Technique
	4.1 Different Versions of the Voltage-Clamp Technique
		4.1.1 The Classic Squid Giant Axon
		4.1.2 The Vaseline- or Sucrose-Gap Voltage Clamp
		4.1.3 The Two-Microelectrode Voltage Clamp
		4.1.4 The One-Electrode Voltage Clamp
		4.1.5 The Open-Oocyte Voltage Clamp
	4.2 Analysing Current Fluctuations
	4.3 Analysing Transient Charge Movements (Gating Currents)
	4.4 The Patch-Clamp Technique
		4.4.1 Different Versions of Patch Clamp (Patch Conformations)
		4.4.2 Advantages of the Different Patch Conformations
		4.4.3 The Single-Channel Current and Conductance
		4.4.4 The Sniffer-Patch Method
	4.5 Automated Electrophysiology
		4.5.1 Automated Patch Clamp
			Take-Home Messages
	References
Chapter 5: Ion-Selective Channels
	5.1 General Characteristics of Ion Channels
		5.1.1 Selectivity of Ion Channels
		5.1.2 Discrete Movement of Ions through Pores
	5.2 Specific Ion Channels
		5.2.1 The Na+ Channel (A Single-Ion Pore)
		5.2.2 The K+ Channel (A Multi-Ion Pore)
		5.2.3 The Ca2+ Channel (A Multi-Ion Pore)
		5.2.4 Anion-Selective Channels
			Take-Home Messages
	References
Chapter 6: Theory of Excitability
	6.1 The Hodgkin-Huxley Description of Excitation
		6.1.1 Experimental Basics
		6.1.2 The Hodgkin-Huxley (HH) Description of Excitability
			6.1.2.1 The Hypothetical Channel
			6.1.2.2 The K+ Channel
			6.1.2.3 The Na+ Channel
			6.1.2.4 The HH Description
		6.1.3 The Action Potential
			6.1.3.1 Phenomenological Description
			6.1.3.2 Calculation of Propagated Action Potential.
	6.2 Continuous and Saltatory Spread of Action Potentials
		6.2.1 The Electrotonic Potential
		6.2.2 The Continuous Spread of an Action Potential
		6.2.3 The Saltatory Spread of an Action Potential
	6.3 Generation and Transmission of Action Potentials
		6.3.1 Generation
		6.3.2 Transmission
	6.4 Summary of the Different Types of Potentials
		6.4.1 Surface Potential
	6.5 Action Potential in Non-nerve Cells
		6.5.1 Skeletal Muscle
		6.5.2 Smooth Muscle
		6.5.3 Heart Muscle
		6.5.4 Plant Cells
			Take-Home Messages
	References
Chapter 7: Carrier-Mediated Transport
	7.1 General Characteristics of Carriers
		7.1.1 Distinction Between Pores and Carriers
		7.1.2 The Oocytes of Xenopus: A Model System
		7.1.3 The Anion Exchanger
		7.1.4 The Sodium Pump
			7.1.4.1 Steady-State Pump Current
			7.1.4.2 Transient Pump-Generated Currents
		7.1.5 The Neurotransmitter Transporter GAT1
	7.2 Carriers Are Like Channels with Alternating Gates
		Take-Home Messages
	References
Chapter 8: Examples of Application of the Voltage-Clamp Technique
	8.1 Structure-Function Relationships of Carrier Proteins
		8.1.1 The Na+,K+-ATPase
		8.1.2 The Na+-Dependent GABA Transporter (GAT1)
	8.2 Structure-Function Relationships of Ion Channels
		8.2.1 Families of Various Ion Channels
			8.2.1.1 The Voltage-Gated Ion Channel (VIC) Superfamily
			8.2.1.2 The Ligand-Gated Ion Channel (LIC) Family
			8.2.1.3 The Chloride Channel (ClC) Family
			8.2.1.4 The Gap Junction-Forming (Connexin) Family
			8.2.1.5 The Epithelial Na+ Channel (ENaC) Family
			8.2.1.6 Mechanosensitive Ion Channels
		8.2.2 ATP-Gated Cation Channel (ACC) Family
			8.2.2.1 Structure and Classification of P2X Receptors
		8.2.3 Experimental Results
			8.2.3.1 The P2X1 Receptor
			8.2.3.2 The P2X2 Receptor
			8.2.3.3 Effect of Glycosylation on P2X1 Receptor Function
	8.3 Viral Ion Channels
		8.3.1 The 3a Protein of SARS Coronavirus
			8.3.1.1 Inhibition of 3a-Mediated Current by the Anthrachinon Emodin
			8.3.1.2 Inhibition of 3a-Mediated Current by the Kaempferol Glycoside Juglanin
		8.3.2 Channel Proteins of SARS Coronavirus-2
		8.3.3 The Viral Protein Unit (Vpu) of HIV-1
		8.3.4 The M2 (Matrix Protein 2) of Influenza a Virus
			8.3.4.1 Inhibition of M2-Mediated Current by Kaempferol Triglycoside
	8.4 Electrophysiology as a Tool in Chinese Medicine Research
		8.4.1 Mechanisms in Acupuncture Points
			8.4.1.1 Mast-Cell Degranulation
			8.4.1.2 Mast-Cell Degranulation Is Initiated by Ion-Channel Activation
		8.4.2 Mechanisms in Effected Sites
			8.4.2.1 Co-Expression of Neurotransmitter Transporters and δ-Opioid Receptor
	8.5 Electrophysiology as a Tool in Pharmacology
		8.5.1 The Na+,Ca2+ Exchanger
		8.5.2 Neurotransmitter Transporters
		8.5.3 Ion Channels
			Take-Home Messages
	References
Chapter 9: Appendix
	9.1 Influence of External Electrical and Magnetic Fields on Physiological Function
		9.1.1 Magnetostatic Fields
		9.1.2 Electrostatic Fields
		9.1.3 Electromagnetic Fields
			9.1.3.1 Low-Frequency Electric Fields (50 Hz)
			9.1.3.2 High Frequency Electric Fields (kHz - GHz)
			9.1.3.3 Conclusion
	9.2 A Laboratory Course: Two-Electrode Voltage Clamp (TEVC)
		9.2.1 Motivation
		9.2.2 Background
			9.2.2.1 Electrical Characteristics of Biological Membranes
				The Membrane Potential
				The Membrane as an Electrical Unit
				Theoretical Background of Voltage Clamp
				The Principle of Voltage Clamp (See Sect. 3.4.5)
				Two-Electrode Voltage Clamp
		9.2.3 Questions to Be Answered for the Course
		9.2.4 Set-up and Basic Instructions
			9.2.4.1 Experimental Set-up (See Fig. 9.6)
			9.2.4.2 Preparation of Microelectrodes
			9.2.4.3 Instructions for the Use of CellWorks Program for the Turbo TEC
			9.2.4.4 Solutions
		9.2.5 Experiments and Data Analysis
			9.2.5.1 IV Characteristics
				Procedure
				Tasks
			9.2.5.2 Hypothesis Testing - the Paired-Sample T-Test
			9.2.5.3 Determination of the Membrane Capacitance
				Procedure
				Tasks
	References
Index