Simultaneous Assessment of cAMP Signaling Events in Different Cellular Compartments Using FRET-Based Reporters -- Recording Intracellular cAMP Levels with EPAC-Based FRET Sensors by Fluorescence Lifetime Imaging -- A Novel Approach Combining Real-Time Imaging and the Patch-Clamp Technique to Calibrate FRET-Based Reporters for cAMP in Their Cellular Microenvironment -- Structure-Based, In Silico Approaches for the Development of Novel cAMP FRET Reporters -- Automated Image Analysis of FRET Signals for Subcellular cAMP Quantification -- Channel-Based Reporters for cAMP Detection -- Imaging Sub-Plasma Membrane cAMP Dynamics with Fluorescent Translocation Reporters -- Adenoviral Transduction of FRET-Based Biosensors for cAMP in Primary Adult Mouse Cardiomyocytes -- Generation of Transgenic Mice Expressing FRET Biosensors -- Photoactivatable Adenylyl Cyclase (PACs) as a Tool to Study cAMP Signaling In Vivo: An Overview -- Selective Disruption of the AKAP Signaling Complexes -- Screening for Small Molecule Disruptors of AKAP-PKA Interactions -- Structure-Based Bacteriophage Screening for AKAP-Selective PKA Regulatory Subunit Variants -- A Yeast-Based High-Throughput Screen for Modulators of Phosphodiesterase Activity -- Separation of PKA and PKG Signaling Nodes by Chemical Proteomics -- Development of Computational Models of cAMP Signaling.

This detailed volume encompasses new technological developments that specifically address questions related to adenosine 3',5'-monophosphate (cAMP) compartmentalization, that probe relevant protein-protein interactions, that increase the spatial and temporal resolution of cAMP signal detection, and that can facilitate integration of the mounting complexity of the information that is becoming available on this signaling system. cAMP, the prototypical intracellular second messenger, regulates a large variety of cellular functions and biological processes, including gene transcription, cell metabolism, proliferation, development, as well as more specialized functions depending on the cell type, so the realization of its extremely complex spatial organization and local regulation is providing novel mechanistic insight into cell physiology and is producing a novel framework for the identification of disease mechanisms. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction 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. Practical and authoritative, cAMP Signaling: Methods and Protocols serves as a vital resource for researchers working in this expanding, complex field.