Nuclear G-Protein Coupled Receptors, 2015 Methods and Protocols Methods in Molecular Biology Series, Vol. 1234

Nuclear G-Protein Coupled Receptors: Methods and Protocols is a compilation of a number of conceptual and methodological aspects important for the validation and characterization of intacrine signaling systems. To date, the best-characterized intracrine signaling system is that of angiotensin II (Ang II), covered in depth in various chapters. Methodology to study the subcellular localization and function of GPCRs and other signaling systems is provided, as well as numerous chapters focusing on methods designed to understand signaling mediated by nuclear and other internal GPCRs. Methods are also described to study the formation of second messengers such as cAMP and to study the trafficking of receptors from the cell surface. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Nuclear G-Protein Coupled Receptors: Methods and Protocols seeks to serve both professionals and novices with state-of-the-art approaches to characterize what is becoming a common theme in cellular signaling.

Studies of Intracellular Angiotensin II.- Single-Cell Microinjection Coupled to Confocal Microscopy to Characterize Nuclear Membrane Receptors in Freshly Isolated Cardiomyocytes.- Design and Application of Light-Activated Probes for Cellular Signaling.- Using Caged Ligands to Study Intracrine Endothelin Signalling In Intact Cardiac Myocytes.- Quantification of Catecholamine Uptake in Adult Cardiac Myocytes.- Characterization of the Interaction Between the Prostaglandin D2 DP1 Receptor and the Intracellular L-Prostaglandin D Synthase.-  Isolation and Study of Cardiac Nuclei from Canine Myocardium and Adult Ventricular Myocytes.- High Resolution Imaging and Function of Nuclear G Protein-Coupled Receptors (GPCRs).- Biochemical Fractionation of Membrane Receptors in the Nucleus.- Functional G Protein-Coupled Receptors on Nuclei From Brain and Primary Cultured Neurons.- Automated Microscopy of Cardiac Myocyte Hypertrophy: A Case Study on the Role of Intracellular a-Adrenergic Receptors.- Measuring Intra-Nuclear and Nuclear Envelope [Ca2+] vs. Cytosolic [Ca2+].- Assessing GPCR and G Protein Signaling to the Nucleus in Live Cells Using Fluorescent Biosensors.- Tandem Affinity Purification to Identify Cytosolic and Nuclear Gbg-Interacting Proteins.- Examining the Effects of Nuclear GPCRs on Gene Expression Using Isolated Nuclei.- Trafficking and Function of GPCRs in the Endosomal Compartment.