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Imaging activity-dependent signaling dynamics at the neuronal synapse using FRET-based biosensors

Item Type:Article
Title:Imaging activity-dependent signaling dynamics at the neuronal synapse using FRET-based biosensors
Creators Name:Farsi, Z. and Woehler, A.
Abstract:In this chapter, we introduce the combined use of FRET-based biosensors and synaptic markers as an effective tool for studying intracellular signaling pathways in small synaptic terminals of neuronal cells. The approach is based on the unmixing of excitation/emission spectral fingerprints of a FRET donor and acceptor pair, as well as a lipophilic styryl dye, FM1-43, loaded into presynaptic terminals. The destaining of FM1-43 during evoked release provides a map to guide the sampling of fluorescence for FRET analysis. In the example presented here, we measure the temporal dynamics of cAMP at the presynaptic terminal using an intramolecular CFP/YFP-based FRET sensor. However, this methodology can be applied to investigate the spatial and temporal regulation of a variety of signaling processes, as well as dynamic changes in protein-protein interaction.
Keywords:FRET, Intracellular Signaling, Live-Cell Imaging, Synaptic Plasticity, CAMP, Animals, Rats
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:Synapse Development : methods and protocols
ISSN:1064-3745
ISBN:978-1-4939-6686-8
Publisher:Springer / Humana Press (U.S.A.)
Volume:1538
Page Range:261-275
Number of Pages:372
Date:2017
Official Publication:https://doi.org/10.1007/978-1-4939-6688-2_18
PubMed:View item in PubMed

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