Patch-Clamp Methods and Protocols (Methods in Molecular Biology, 1183)

Since its inception, patch-clamp has continued to be widely considered the gold standard method to record ion channel activity. __Patch-Clamp Methods and Protocols, Second Edition__, provides a comprehensive collection of new techniques for the development of automated, high-throughput screening systems for pharmacological evaluation, the use of various patch-clamp configurations together with novel molecular biological and imaging methodologies and enhanced stimulation protocols and perfusion systems. Divided into sections on pharmacology, physiology and biophysics, the chapters cover methods to generate more physiologically relevant conditions for drug application and screening technologies, recently developed applications such as optogenetic stimulation, advances in whole-cell recordings in freely-moving animals and novel technologies to create custom microelectrodes designed for reducing the access resistance and improving the rate of molecular diffusion. Patch-clamp is an indispensable technique for conducting pharmacological, physiological and biophysical research aimed at understanding crucial aspects of cellular and network function. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, __Patch-Clamp Methods and Protocols, Second Edition__ will provide a useful technical and methodological guide to diverse audiences of electrophysiologists, from students to experienced investigators. 1. History of electrophysiology and the patch clamp / Alexei Verkhratsky and Vladimir Parpura 2. Whole-cell patch-clamp analysis of recombinant NMDA receptor pharmacology using brief glutamate applications / Nathan G. Glasgow and Jon W. Johnson 3. Patch clamp combined with voltage/concentration clamp to determine the kinetics and voltage dependency on N-methyl-D-aspartate (NMDA) receptor open channel blockers / Chris G. Parsons and Kate E. Gilling 4. State-of-the-art automated patch clamp: heat activation, action potentials, and high throughput in ion channel screening / Sonja Stoelzle-Feix 5. Industrializing electrophysiology: HT automated patch clamp on SyncroPatch96 using instant frozen cells / Liudmila Polonchuk 6. Planar patch clamp for neuronal networks - considerations and future perspectives / Alessandro Bosca, Marzia Martina, and Christophe Py 7. Methods for the study of synaptic receptor functional properties / Enrica Maria Petrini and Andrea Barberis 8. Single cell RT-PCR, a technique to decipher the electrical, anatomical, and genetic determinants of neuronal diversity / Maria Toledo-Rodriguez and Henry Markham 9. Whole-cell voltage clamp on skeletal muscle fibers with the silicone-clamp technique / Romain Lefebvre, Sandrine Pouvreau, Claude Collet, Bruno Allard, and Vincent Jacquemond 10. Multiphoton imaging approaches for studying striatal dendritic excitability / Joshua L. Plotkin and D. James Surmeier 11. Investigation of synaptic microcircuits using patch-clamp paired recordings in acute brain slices / Giovanni Russo and Stefano taverna 12. Outside-out "sniffer-patch" clamp technique for in situ measures of neurotransmitter release / Émilie Muller-Chrétien 13. A cost-effective method for preparing, maintaining, and transfecting neurons in organotypic slices / Cary Soares, Kevin F.H. Lee, Denise Cook and Jean-Claude Béïque 14. Acute brain slice methods for adult and aging animals: application of targeted patch clamp analysis and optogenetics / Jonathan T. Ting, Tanya L. Daigle, Qian Chen and Guoping Feng 15. Synaptic connectivity in engineered neuronal networks / Peter Molnar, Jung-Fong Kang, Neelima Bhargava, Mainak Das and James J. Hickman 16. Modeling of action potential generation in NG108-15 cells / Peter Molnar and James J. Hickman 17. Whole-cell patch-clamp recordings in freely moving animals / Albert K. Lee, Jâerãome Epsztein and Michael Brecht 18. Pressure-polished borosilicate pipettes are "universal sealer" yielding low access resistance and efficient intracellular perfusion / Marco Aquila, Mascia Benedusi, Anna Fasoli and Giorgio Rispoli 19. Current recordings at the single channel level in adult mammalian isolated cardiomyocytes / Romain Guinamard, Thomas Hof, and Laurent Sallâe 20. Dynamic clamp as a tool to study the functional effects of individual membrane currents / Géza Berecki, Arie O. Verkerk, Antoni C.G. van Ginneken and Ronald Wilders 21. Dynamic clamp in cardiac and neuronal systems using RTXI / Francis A. Ortega, Robert J. Butera, David J. Christini, John A. White and Alan D. Dorval II 22. Ion selectivity of pore-forming peptides and ion channels measured in xenopus oocytes / Thierry Cens and Pierre Charnet 23. Principles of single-channel kinetic analysis / Feng Qin. Since its inception, patch-clamp has continued to be widely considered the gold standard method to record ion channel activity. Patch-Clamp Methods and Protocols, Second Edition, provides a comprehensive collection of new techniques for the development of automated, high-throughput screening systems for pharmacological evaluation, the use of various patch-clamp configurations together with novel molecular biological and imaging methodologies, and enhanced stimulation protocols and perfusion systems. Divided into sections on pharmacology, physiology, and biophysics, the chapters cover methods to generate more physiologically relevant conditions for drug application and screening technologies, recently developed applications such as optogenetic stimulation, advances in whole-cell recordings in freely-moving animals, and novel technologies to create custom microelectrodes designed for reducing the access resistance and improving the rate of molecular diffusion. Patch-clamp is an indispensable technique for conducting pharmacological, physiological and biophysical research aimed at understanding crucial aspects of cellular and network function. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Patch -Clamp Methods and Protocols, Second Edition will provide a useful technical and methodological guide to diverse audiences of electrophysiologists, from students to experienced investigators Front Matter....Pages i-xi Front Matter....Pages 1-19 Front Matter....Pages 21-21 Front Matter....Pages 23-41 Front Matter....Pages 43-63 Back Matter....Pages 65-80 ....Pages 81-92