Freeze-frame imaging of synaptic activity using SynTagMA

• Published in: Nature communications Vol. 11; no. 1; p. 2464
• Main Authors: Perez-Alvarez, Alberto, Fearey, Brenna C., O’Toole, Ryan J., Yang, Wei, Arganda-Carreras, Ignacio, Lamothe-Molina, Paul J., Moeyaert, Benjamien, Mohr, Manuel A., Panzera, Lauren C., Schulze, Christian, Schreiter, Eric R., Wiegert, J. Simon, Gee, Christine E., Hoppa, Michael B., Oertner, Thomas G.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 18.05.2020; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 14; 14/10; 14/35; 14/69; 631/378/87; 631/443/376; 64/60; 64/86; Action Potentials; Animals; Axon guidance; Axons; Axons - metabolism; Biomarkers - metabolism; Brain; Brain research; Brain slice preparation; Calcium; Calcium imaging; Cell culture; Cells, Cultured; Female; Fluorescence; Gene mapping; Genetic code; Hippocampus - cytology; Humanities and Social Sciences; Imaging, Three-Dimensional; Light; Male; Mapping; Mice, Inbred C57BL; Microscopy; multidisciplinary; Neuroimaging; Neurons; Neurons - metabolism; Presynaptic Terminals - metabolism; Rats, Sprague-Dawley; Rats, Wistar; Science; Science (multidisciplinary); Synapses; Synapses - physiology; Synaptophysin - metabolism; Time Factors; Windows (intervals)
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-16315-4

Information within the brain travels from neuron to neuron across billions of synapses. At any given moment, only a small subset of neurons and synapses are active, but finding the active synapses in brain tissue has been a technical challenge. Here we introduce SynTagMA to tag active synapses in a user-defined time window. Upon 395–405 nm illumination, this genetically encoded marker of activity converts from green to red fluorescence if, and only if, it is bound to calcium. Targeted to presynaptic terminals, preSynTagMA allows discrimination between active and silent axons. Targeted to excitatory postsynapses, postSynTagMA creates a snapshot of synapses active just before photoconversion. To analyze large datasets, we show how to identify and track the fluorescence of thousands of individual synapses in an automated fashion. Together, these tools provide an efficient method for repeatedly mapping active neurons and synapses in cell culture, slice preparations, and in vivo during behavior. Calcium imaging has been used to visualize the activity of individual synapses, but cannot be scaled up to monitor thousands of synapses in tissue. Here, the authors present genetic tools that can be photoconverted from green to red to create a map of active synapses.