Real-time monitoring of cyclic nucleotide signaling in neurons using genetically encoded FRET probes
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Signaling cascades involving cyclic nucleotides play key roles in signal transduction in virtually all cell types. Elucidation of the spatiotemporal regulation of cyclic nucleotide signaling requires methods for tracking the dynamics of cyclic nucleotides and the activities of their regulators and effectors in the native biological context. Here we review a series of genetically encoded FRET-based probes for real-time monitoring of cyclic nucleotide signaling with a particular focus on their implementation in neurons. Current data indicate that neurons have a very active metabolism in cyclic nucleotide signaling, which is tightly regulated through a variety of homeostatic regulations.
KeywordsForskolin Brain Slice Cyclic Nucleotide Guanylyl Cyclase cAMP Concentration
We would like to thank Professor Ron Harris-Warrick for critically reading this manuscript. This work was supported by CNRS, UPMC, “Fondation pour la Recherche Médicale” and “Fondation pour la Recherche sur le Cerveau” (to P. V.); and by NIH (DK073368 and CA122673), the American Heart Association, the Young Clinical Scientist Award Program of the Flight Attendant Medical Research Institute, and 3M (to J. Z.).
- Domingo, B. et al. (2007). Imaging FRET standards by steady-state fluorescence and lifetime methods. Microsc. Res. Tech. 70, 1010–1021Google Scholar
- Rich, T. C. et al. (2001a). In vivo assessment of local phosphodiesterase activity using tailored cyclic nucleotide-gated channels as cAMP sensors. J. Gen. Physiol. 118, 63–78Google Scholar
- Rich, T. C. et al. (2001b). A uniform extracellular stimulus triggers distinct cAMP signals in different compartments of a simple cell. Proc. Natl. Acad. Sci. USA 98, 13049–13054Google Scholar
- Sato, M., Hida, N., and Umezawa, Y. (2005). Imaging the nanomolar range of nitric oxide with an amplifier-coupled fluorescent indicator in living cells. Proc. Natl. Acad. Sci. USA 102, 14515–14520Google Scholar
- Saucerman, J. J. et al. (2006) Systems analysis of PKA-mediated phosphorylation gradients in live cardiac myocytes. Proc. Natl. Acad. Sci. USA, 19, 2650–2658Google Scholar