Gap Junction Protocols

Presenting state-of-the-art protocols to study gap junctions, this detailed book first focuses on the use of methods and tools to investigate the different aspects of connexin expression and gap junction regulation. The second part of the volume describes several methods to probe gap junction functionality as such. 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 and readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Gap Junction Protocols is intended for basic and applied researchers in the area of biomedical and life sciences, both in academic and industrial settings.

Includes cutting-edge methods and protocols involving gap junctions Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Analysis of Liver Connexin Expression Using Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction.- DNA Methylation Analysis of Human Tissue-Specific Connexin Genes.- Detection of Connexins in Liver Cells Using Sodiumdodecylsulfate Polyacrylamide Gel Electrophoresis and Immunoblot Analysis.- Immunohisto- and Cytochemistry Analysis of Connexins.- Small Interfering RNA-Mediated Connexin Gene Knockdown in Vascular Endothelial and Smooth Muscle Cells.- Generation and Use of Trophoblast Stem Cells and Uterine Myocytes to Study the Role of Connexins for Pregnancy and Labor.- Identification of Connexin43 Phosphorylation and S -Nitrosylation in Cultured Primary Vascular Cells.- Preparation of Gap Junctions in Membrane Microdomains for Immunoprecipitation and Mass Spectrometry Interactome Analysis.- Scrape Loading/Dye Transfer Assay.- Microinjection Technique for Assessment of Gap Junction Function.- Electroporation Loading and Dye Transfer: A Safe and Robust Method to Probe Gap Junctional Coupling.- Using Fluorescence Recovery After Photobleaching to Study Gap Junctional Communication In Vitro.- Tracking Dynamic Gap Junctional Coupling in Live Cells by Local Photo-Activation and Fluorescence Imaging.- A Functional Assay to Assess Connexin43-Mediated Cell-to-Cell Communication of Second Messengers in Cultured Bone Cells.- Calcium Wave Propagation Triggered by Local Mechanical Stimulation as a Method for Studying Gap Junctions and Hemichannels.- Establishment of the Dual Whole Cell Recording Patch Clamp Configuration for the Measurement of Gap Junction Conductance.