Nuclear G-Protein Coupled Receptors: Methods and Protocols

ISBN-13: 9781493917549 / Angielski / Twarda / 2014 / 215 str.

Bruce G. Allen; Terence E. Hebert

Opis

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.

Spis treści

1. Studies of Intracellular Angiotensin II

Richard N. Re and Julia L. Cook

 

2. Single-Cell Microinjection Coupled to Confocal Microscopy to Characterize Nuclear Membrane Receptors in Freshly Isolated Cardiomyocytes

Clémence Merlen and Jonathan Ledoux

 

3. Design and Application of Light-Activated Probes for Cellular Signaling

David Chatenet, Steve Bourgault, and Alain Fournier

 

4. Using Caged Ligands to Study Intracrine Endothelin Signalling In Intact Cardiac Myocytes

Clémence Merlen, Louis R. Villeneuve, and Bruce G. Allen

 

5. Quantification of Catecholamine Uptake in Adult Cardiac Myocytes

Erika F. Dahl, Casey D. Wright, and Timothy D. O’Connell

 

6. Characterization of the Interaction Between the Prostaglandin D2 DP1 Receptor and the Intracellular L-Prostaglandin D Synthase      

Chantal Binda and Jean-Luc Parent

 

7.  Isolation and Study of Cardiac Nuclei from Canine Myocardium and Adult Ventricular Myocytes

Artavazd Tadevosyan, Bruce G. Allen, and Stanley Nattel

 

8. High Resolution Imaging and Function of Nuclear G Protein-Coupled Receptors (GPCRs)

Vikrant K. Bhosle, Fernand Gobeil Jr., Jose Carlos Rivera, Alfredo Ribeiro-da-Silva, and Sylvain Chemtob

 

9. Biochemical Fractionation of Membrane Receptors in the Nucleus

Ying-Nai Wang, Longfei Huo, Jennifer L. Hsu, and Mien-Chie Hung

 

10. Functional G Protein-Coupled Receptors on Nuclei From Brain and Primary Cultured Neurons

Yuh-Jiin I. Jong and Karen L. O’Malley

 

11. Automated Microscopy of Cardiac Myocyte Hypertrophy: A Case Study on the Role of Intracellular a-Adrenergic Receptors

Karen A. Ryall and Jeffrey J. Saucerman

 

12. Measuring Intra-Nuclear and Nuclear Envelope [Ca2+] vs. Cytosolic [Ca2+]

Senka Ljuboyević and Donald M. Bers

 

13. Assessing GPCR and G Protein Signaling to the Nucleus in Live Cells Using Fluorescent Biosensors

Julie Bossuyt and Donald M. Bers

 

14. Tandem Affinity Purification to Identify Cytosolic and Nuclear Gbg-Interacting Proteins

Rhiannon Campden, Darlaine Pétrin, Mélanie Robitaille, Nicolas Audet, Sarah Gora, Stéphane Angers, and Terence E. Hébert

 

15. Examining the Effects of Nuclear GPCRs on Gene Expression Using Isolated Nuclei

George Vaniotis, Sarah Gora, André Nantel, Terence E. Hébert, and Bruce G. Allen

 

16. Trafficking and Function of GPCRs in the Endosomal Compartment

Davide Calebiro, Amod Godbole, Sandra Lyga, and Martin J. Lohse