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Full Description
This volume explores the latest developments in the study of connexin hemichannels (HCs). The chapters in this book cover topics such as purification, reconstitution, and functional analysis of connexin HCs; spatial and temporal localization of connexins in cells using confocal microscopy; evaluation of connexin HC activity in vivo; generation of connexin-expressing stable cell pools; and automated assessment of cutaneous pathology in a mouse model of HC dysfunction. 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.
Cutting-edge and comprehensive, Connexin Hemichannels: Methods and Protocols is a valuable resource that discusses the importance of HCs as drug targets, and how studying them is essential to identifying newcompounds capable of modulating HC function selectively and specifically.
Contents
Purification, Reconstitution, and Functional Analysis of Connexin Hemichannels.- Expression and Function of Connexins in Extracellular Vesicles.- Expression, Purification, and Nanodisc Reconstitution of Connexin-43 Hemichannels for Structural Characterization by Cryo-Electron Microscopy.- Molecular Dynamics Simulation of Permeation through Connexin Channels.- Spatial and Temporal Localization of Connexins in Cells Using Confocal Microscopy.- Assessment of Connexin43 Hemichannel Functionality Based on Cytosolic Uptake of Yo-Pro1.- A Dye Uptake Assay to Measure Large-Pore Channel Activity in Trypanosoma cruzi.- Measurement of Ca2+ Uptake through Connexin Hemichannels.- Evaluation of Connexin Hemichannel Activity In Vivo.- Protocol for the Study of Connexin and DNA Interactions.- Protocol for the Electrophysiological Characterization of Endogenous Pannexin 1 Intercellular Cell-Cell Channels.- Generation of Connexin-Expressing Stable Cell Pools.- A Protocol for the Automated Assessment of Cutaneous Pathology in a Mouse Model of Hemichannel Dysfunction.- Measuring Connexin Hemichannel Opening in Response to a InsP3-Mediated Cytosolic Ca2+ Increase.
