Chemical Neurobiology: Methods and Protocols

ISBN-13: 9781627033442 / Angielski / Twarda / 2013 / 202 str.

Matthew Banghart

Opis

Many advances in modern neuroscience are enabled by the availability of chemical tools that allow sensitive, precise, and quantitative measurements of, and control over, biological processes. These powerful reagents are widely used for investigating the nervous system at levels of detail ranging from ion channel structure to neural network dynamics. Recent advances in photochemistry, microscopy, and protein engineering have triggered a surge in the development and application of these interdisciplinary techniques. Chemical Neurobiology: Methods and Protocols assists with the design, characterization and validation of new chemical tools for neurobiology by providing detailed protocols of procedures and assays deemed essential for the successful development and implementation of such tools. Divided into three sections, topics cover chemical probes of membrane protein structure and function, photochemical control of protein and cellular function, and chemical probes for imaging in the nervous system. 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, Chemical Neurobiology: Methods and Protocols serves scientists at many levels, including students aiming to expand their perspective, laboratory researchers seeking technical guidance, and established investigators looking for creative solutions to their research problems in molecular, cellular and systems neuroscience.

Spis treści

Part I. Chemical Probes of Membrane Protein Structure and Function

 

1. Engineering K+ Channels using Semisynthesis

Alexander G. Komarov, Corey A. Costantino, and Francis I. Valiyaveetil

 

2. Chemical Derivatization and Purification of Peptide-Toxins for Probing Ion Channel Complexes

Zhengmao Hua and William R. Kobertz

 

3. Using Yeast to Study Potassium Channel Function and Interactions with Small Molecules

Sviatoslav N. Bagriantsev and Daniel L. Minor Jr.

 

4. A FLIPR Assay for Evaluating Agonists and Antagonists of GPCR Heterodimers

Jessica H. Harvey, Richard M. van Rijn, and Jennifer L. Whistler

 

Part II. Photochemical Control of Protein and Cellular Function

 

5. Characterizing Caged Molecules Through Flash Photolysis and Transient Absorption Spectroscopy

Joseph P. Y. Kao and Sukumaran Muralidharan

 

6. Characterization of One- and Two- Photon Photochemical Uncaging Efficiency

Alexandre Specht, Frederic Bolze, Jean Francois Nicoud, and Maurice Goeldner

 

7. Photochromic Potassium Channel Blockers: Design and Electrophysiological Characterization

Alexandre Mourot, Timm Fehrentz, and Richard H. Kramer

 

8. A 1H NMR Assay for Measuring the Photostationary States of Photoswtichable Ligands

Matthew R. Banghart and Dirk Trauner

 

9. Developing a Photoreactive Antagonist

Pamela M. England

 

Part III. Chemical Probes for Imaging in the Nervous System

 

10. Development and in vitro Characterization of Ratiometric and Intensity-based Fluorescent Ion Sensors

Laurel A. Cooley, Vladimir V. Martin, and Kyle R. Gee

 

11.  Characterization of Voltage-Sensitive Dyes in Living Cells using Two-Photon Excitation

Corey D. Acker and Leslie M. Loew

           

12. Characterization and Validation of Fluorescent Receptor Ligands: A Case Study of the Ionotropic Serotonin Receptor

Ruud Hovius

 

13.  Imaging Single Synaptic Vesicles in Mammalian Central Synapses with Quantum Dots

Qi Zhang

 

14.  Directed Evolution of Protein-Based Neurotransmitter Sensors for MRI 

Philip A. Romero, Mikhail G. Shapiro, Frances H. Arnold, and Alan Jasanoff