Connexin Methods and Protocols

Tools to Study Connexins.- Investigation of Connexin Gene Expression Patterns by In Situ Hybridization Techniques.- Sodium Dodecyl Sulfate-Freeze-Fracture Immunolabeling of Gap Junctions.- Purification of Gap Junctions.- Culturing of Mammalian Cells Expressing Recombinant Connexins and Two-Dimensional Crystallization of the Isolated Gap Junctions.- Connexins/Connexons.- Biochemical Analysis of Connexon Assembly.- Expression and Imaging of Connexin-GFP Chimeras in Live Mammalian Cells.- Analysis of Connexin Expression in Brain Slices by Single-Cell Reverse Transcriptase Polymerase Chain Reaction.- Use of Retroviruses to Express Connexins.- Spatiotemporal Depletion of Connexins Using Antisense Oligonucleotides.- Transfection and Expression of Exogenous Connexins in Mammalian Cells.- Assays For Function.- Assaying the Molecular Permeability of Connexin Channels.- Applying the Xenopus Oocyte Expression System to the Analysis of Gap Junction Proteins.- Mutagenesis to Study Channel Structure.- Dual Patch Clamp.- Determining Ionic Permeabilities of Gap Junction Channels.- Fluorescence Recovery After Photobleaching.- Capture of Transjunctional Metabolites.- Physiology and Biology of Connexins.- The Study of Connexin Hemichannels (Connexons) in Xenopus Oocytes.- Exploring Hemichannel Permeability In Vitro.- Inducing De Novo Formation of Gap Juntion Channels.- Recording and Analysis of Putative Direct Electrical Interactions in the Mammalian Brain.- Intercellular Calcium Signaling and Flash Photolysis of Caged Compounds.- Biochemical Analysis of Connexin Phosphorylation.- How to Close a Gap Junction Channel.

"This book will be invaluable to those in the connexin field and a fascinating read and useful text to those new to this area of research." -Molecular Biotechnology