Main description:

Recently, many ground-breaking steps have been made towards better understanding NO/cGMP/PKG pathways, its components, substrates, and their localization within a given cell. These advances were only possible due to the development of sophisticated new techniques in the field. In Guanylate Cyclase and Cyclic GMP: Methods and Protocols expert researchers in the field seek to provide an overview of novel techniques to identify various elements of the NO/cGMP/PKG pathway and further characterize their function, signaling, localization, and importance on a cellular level and in whole animal models providing a higher patho-/physiological integration and relevance. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Guanylate Cyclase and Cyclic GMP: Methods and Protocols seeks to provide scientist current methods and a useful guide towards the possibility to apply these techniques to their own research.

Contents:

1.NO-cGMP: The Past, the Present and the Future

Michael Russwurm, Corina Russwurm, Doris Koesling, and Evanthia Mergia

2. cGMP Dependent Protein Kinases (cGK)Franz Hofmann and Joerg W. Wegener

3. Enzyme Assays for cGMP Hydrolysing Phosphodiesterases

Sergei D. Rybalkin, Thomas R. Hinds and Joseph A. Beavo

4. Radioimmunoassay for the Quantification of cGMP Levels in Cells and Tissues Ronald Jager, Dieter Groneberg, Barbara Lies, Noomen Bettaga, Michaela Kummel, and Andreas Friebe5. Hyperspectral Imaging of FRET-based cGMP Probes

Thomas C. Rich, Andrea L. Britain, Tiffany Stedman, and Silas J. Leavesley

6. Visualization of cGMP with cGi Biosensors

Martin Thunemann, Natalie Fomin, Christian Krawutschke, Michael Russwurm, and Robert Feil

7. Advances and techniques to measure cGMP in intact cardiomyocytesKonrad R. Goetz and Viacheslav O. Nikolaev

8. Real-time Monitoring the Spatio-temporal Dynamics of Intracellular cGMP in Vascular Smooth Muscle CellsKara F. Held and Wolfgang R. Dostmann

9. Methods for Identification of cGKI Substrates

Katja Salb and Jens Schlossmann

10. Approaches for Monitoring PKG1a Oxidative Activation

Joeseph R. Burgoyne and Philip Eaton

11. Analysis of cGMP Signaling in Adipocytes

Katja Jennissen, Bodo Haas, Michaela M. Mitschke, Franziska Siegel and Alexander Pfeifer

12. A Genetic Strategy for the Analysis of Individual Axon Morphologies in cGMP Signalling Mutant Mice

Hannes Schmidt, Gohar Ter-Avetisyan and Fritz G. Rathjen

13. Receptor Binding Assay for NO-Independent Activators of soluble Guanylate Cyclase

Peter M. Schmidt and Johannes-Peter Stasch

14. Direct Intrathecal Drug Delivery in Mice for Detecting In Vivo Effects of cGMP on Pain Processing

Ruirui Lu and Achim Schmidtko

15. The Geisler Method: Tracing Activity Dependent cGMP Plasticity Changes Upon Double Detection of mRNA and Protein on Brain Slices Wibke Singer, Hyun-Soon Geisler and Marlies Knipper

16. Detection of cGMP in the Degenerating Retina

Stylianos Michalakis, Jianhua Xu and Martin Biel, Xi-Qin Ding