Ion Channels: Methods and Protocols

ISBN-13: 9781493960095 / Angielski / Miękka / 2016 / 440 str.

Nikita Gamper

Opis

This Methods in Molecular Biology (TM) book offers strategies and protocols for studying a large group of proteins that form ionic channels in the plasma membrane and intracellular membranes of cells. Includes step-by-step protocols, materials lists, tips and more.

Spis treści

Part I: Cloning and Heterologous Expression of Ion Channel Genes

 

1. Approaches to Cloning of Pain-Related Ion Channel Genes

            Armen N. Akopian

 

2. Mammalian Expression Systems and Transfection Techniques

            Daunia Laurenti and Lezanne Ooi

 

3. Use of Escherichia coli for the Production and Purification of Membrane Proteins

            Vincent G.L. Postis, Andrea E. Rawlings, Amelia Lesiuk, and Stephen A. Baldwin

 

4. Transient Overexpression of Genes in Neurons Using Nucleofection

            Hannah M. Kirton, Louisa Pettinger, and Nikita Gamper

 

5. Viral Gene Delivery: Optimized Protocol for Production of High Titre Lentiviral Vectors

            James Hewinson, Julian F.R. Paton, and Sergey Kasparov

 

Part II: Electrophysiological Methods to Study Ion Channel Functions

 

6. Two-Electrode Voltage Clamp

            Bingcai Guan, Xingjuan Chen, and Hailin Zhang

 

7. Conventional Micropipette-Based Patch Clamp Techniques

            Jonathan D. Lippiat and David C. Wrighton

 

8. Recording of Ion Channel Activity in Planar Lipid Bilayers Experiments

            Eleonora Zakharian

 

9. Recording Macroscopic Currents in Large Patches from Xenopus Oocytes

            Tibor Rohacs

 

10. Combined Single-Channel and Macroscopic Recording Techniques to Analyze Gating Mechanisms of the Large Conductance Ca²⁺ and Voltage Activated (BK) Potassium Channel

            Nguyen V. Nguyen, Aleksandra Gruslova, Wojciech A. Kosiba, and Bin Wang

 

11. Perforated Whole Cell Patch Clamp Recording

            John E. Linley

 

12. Piezo-Electrically Driven Mechanical Stimulation of Sensory Neurons

            Jizhe Hao, Jérôme Ruel, Bertrand Coste, Yann Roudaut, Marcel Crest, and Patrick Delmas

 

13. Automated Planar Patch-Clamp

            Carol J. Milligan and Clemens Möller

 

14. Recording Single-Channel Currents Using “Smart Patch-Clamp” Technique

            Anamika Bhargava and Julia Gorelik

 

Part III: Imaging and Fluorescence Methods to Study Ion Channels

 

15. Using Total Internal Reflection Fluorescence Microscopy to Observe Ion channel Trafficking and Assembly

            Sarah Schwarzer, Gregory I. Mashanov, Justin E. Molloy, and Andrew Tinker

 

16. Förster Resonance Energy Transfer-Based Imaging at the Cell Surface of Live Cells

            Sonya M. Bierbower and Mark S. Shapiro

 

17. The Use of Dansyl-Calmodulin to Study Interactions with Channels and Other Proteins

            Alessandro Alaimo, Covadonga Malo, Pilar Areso, Kerman Aloria, Oscar Millet, and Alvaro Villarroel

 

18. Imaging and Quantification of Recycled K_ATP Channels

            Christopher J. Cockcroft

 

Part IV: Biochemical and Structure-Functional Approaches in Ion Channel Studies

 

19. Generation of Antibodies that Are Externally-Acting Isoform-Specific Inhibitors of Ion Channels

            Jacqueline Naylor and David J. Beech

 

20. Site-Directed Mutagenesis to Study the Structure-Function Relationships of Ion Channels

            Wei Yang and Lin-Hua Jiang

 

21. Cysteine-Based Cross-Linking Approach to Study Inter-Domain Interactions in Ion Channel Functions

            Lin-Hua Jiang

 

22. Analysis of Ca²⁺-Binding Sites in the MthK RCK Domain by X-Ray Crystallography

            Frank J. Smith and Brad S. Rothberg

 

23. Isotope Labeling Strategies for Analysis of an Ion Channel Cytoplasmic Domain by NMR Spectroscopy

            Karin Abarca-Heidemann, Elke Duchardt-Ferner, Jens Woehnert, and Brad S. Rothberg

 

Part V: Studying Ion Channels in Native Tissues

 

24. Recording Dendritic Ion Channel Properties and Function from Cortical Neurons

            Mala M. Shah

 

25. M Current Recording from Acute DRG Slices

            Kirstin E. Rose, Sylvain Gigout, and Nikita Gamper

 

26. Studying Ion Channels in Human Erythrocytes by Direct and Indirect Means

            Stephan M. Huber

 

27. Recording Ion Channels in Isolated, Split-Opened Tubules

            Elena Mironova, Vladislav Bugay, Oleh Pochynyuk, Alexander Staruschenko, and James D. Stockand

 

28. Single Channel Analysis of TRPC Channels in the Podocytes of Freshly Isolated Glomeruli

            Daria V. Ilatovskaya and Alexander Staruschenko

 

29. Ca²⁺ Imaging as a Tool to Assess TRP Channel Function in Murine Distal Nephrons

            Mykola Mamenko, Oleg Zaika, Roger G. O’Neil, and Oleh Pochynyuk

 

30. Patch Clamping Drosophila Sensory Neurons

            Volodymyr Kucher, Benjamin A. Eaton, James D. Stockand, and Nina Boiko

 

Part VI: Ion Channels as Research Tools

 

31. Production and Validation of Recombinant Adeno-Associated Virus for Channelrhodopsin Expression in Neurons

            John Y. Lin

 

32. Optical Control of Ligand-Gated Ion Channels

            Stephanie Szobota, Catherine McKenzie, and Harald Janovjak

Blurb

Every cell in our bodies contains a great variety and number of permeability pathways for various organic and inorganic ions, water, metabolites, nutrients, and signalling molecules. Maintenance and precise control of gating within these pathways are fundamental principles of life as these underlie basic cellular functions such as communication, contractility, and metabolism.  In Ion Channels: Methods and Protocols, Second Edition, experts in the field contribute chapters that focus on the strategies, approaches, methods, and protocols for studying a large family of proteins that form ionic channels in the plasma membrane and intracellular membranes of cells. Using practical examples from the cutting-edge current research, this volume will take a look back at the major methods and approaches that aided the progress toward the current understanding of ion channel function, structural design, and biological roles. The volume also aims to look forward and identify approaches that will lead us to future discoveries. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

 

Authoritative and cutting-edge, Ion Channels: Methods and Protocols, Second Edition will greatly assist researchers searching for specific methodology in studying ion channels on the path toward a greater understanding of these key biological features.