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This volume discusses the latest techniques used by researchers to measure cAMP activity at the cell population, whole cell, and subcellular level. The techniques covered in the book address questions related to cAMP compartmentalization, which look at relevant protein-protein interactions that increase the spatial and temporal resolution of cAMP signals detection, and that can help in the integration of the increasingly complex information that is becoming available in this field. 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, cAMP Signaling: Methods and Protocols, Second Edition is a valuable resource for scientists and researchers who are interested in learning more about this important and developing field.
Chapter 7 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Inhaltsverzeichnis
Real-Time Measurements of Intracellular cAMP Gradients using FRET-Based cAMP Nanorulers.- Assaying Protein Kinase A Activity using a FRET-Based Sensor Purified from Mammalian Cells.- MultiFRET: A Detailed Protocol for High-Throughput Multiplexed Ratiometric FRET.- Photoactivated Adenylyl Cyclases as Optogenetic Modulators of Neuronal Activity.- Imaging the cAMP Signaling Microdomain of the Primary Cilium using Targeted FRET-Based Biosensors.- Methods to Assess Phosphodiesterase and/or Adenylyl Cyclase Activity via Heterologous Expression in Fission Yeast.- Time-Domain Fluorescence Lifetime Imaging of cAMP Levels with EPAC-Based FRET Sensors.- Disruptors of AKAP-Dependent Protein-Protein Interactions.- Micro-2D Cell Culture for cAMP Measurements using FRET Reporters in Human iPSC-Derived Cardiomyocytes.- Automated Image Analysis of FRET Signals for Subcellular cAMP Quantification.- In Vivo cAMP Dynamics in Drosophila Larval Neurons.- Live Cell Imaging of Cyclic Nucleotides in Human Cardiomyocytes.- Optogenetic Control of Heart Rhythm: Lightly Guiding the Cardiac Pace.- Live Imaging of cAMP Signaling in D. discoideum Based on a Bright Bioluminescent Protein, Nano-Lantern.- Generation of Transgenic Mice Expressing Cytosolic and Targeted FRET Biosensors for cAMP and cGMP.- How to Make the CUTiest Sensor in 3 Simple Steps for Computational Pedestrians.- Ion-Channel Based Reporters for cAMP Detection.- Quantitative Phosphoproteomics to Study cAMP Signalling.- Biochemical Analysis of AKAP-Anchored PKA Signaling Complexes.- Fluorescent Translocation Reporters for Sub-Plasma Membrane cAMP Imaging.- A Live-Cell Imaging Assay for Nuclear Entry of cAMP-Dependent Protein Kinase Catalytic Subunits Stimulated by Endogenous GPCR Activation.- Measuring Spatiotemporal cAMP Dynamics within an Endogenous Signaling Compartment using FluoSTEP-ICUE.
Zusammenfassung
This volume discusses the latest techniques used by researchers to measure cAMP activity at the cell population, whole cell, and subcellular level.
