Abstract
Over the last years, the analysis of synaptic connectivity in the mammalian brain has been accelerated by the use of techniques combining electrophysiology, light microscopy, viral tracing, and genetic manipulations in animal models. Of particular interest are methods that aim to label synapses by tethering complementary split GFP fragments in opposing sites of the synaptic cleft. Here, I describe SynView, a method for monitoring synapse formation based on GFP complementation, and provide a detailed protocol for use in neuronal cultures from mouse hippocampus.
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References
Helmstaedter M, Briggman KL, Turaga SC, Jain V, Seung HS, Denk W (2013) Connectomic reconstruction of the inner plexiform layer in the mouse retina. Nature 500:168–174
Wickersham IR, Lyon DC, Barnard RJ, Mori T, Finke S, Conzelmann KK, Young JA, Callaway EM (2007) Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons. Neuron 53:639–647
Callaway EM, Luo L (2015) Monosynaptic circuit tracing with glycoprotein-deleted rabies viruses. J Neurosci 35:8979–8985
Beier KT, Steinberg EE, DeLoach KE, Xie S, Miyamichi K, Schwarz L, Gao XJ, Kremer EJ, Malenka RC, Luo L (2015) Circuit architecture of VTA dopamine neurons revealed by systematic input–output mapping. Cell 162:622–634
Sudhof TC (2008) Neuroligins and neurexins link synaptic function to cognitive disease. Nature 455:903–911
Yamagata M, Sanes JR (2012) Transgenic strategy for identifying synaptic connections in mice by fluorescence complementation (GRASP). Front Mol Neurosci 5:18
Feinberg EH, Vanhoven MK, Bendesky A, Wang G, Fetter RD, Shen K, Bargmann CI (2008) GFP Reconstitution Across Synaptic Partners (GRASP) defines cell contacts and synapses in living nervous systems. Neuron 57:353–363
Kim J, Zhao T, Petralia RS, Yu Y, Peng H, Myers E, Magee JC (2012) mGRASP enables mapping mammalian synaptic connectivity with light microscopy. Nat Methods 9:96–102
Tsetsenis T, Boucard AA, Arac D, Brunger AT, Sudhof TC (2014) Direct visualization of trans-synaptic neurexin-neuroligin interactions during synapse formation. J Neurosci 34:15083–15096
Araç D, Boucard AA, Ozkan E, Strop P, Newell E, Südhof TC, Brunger AT (2007) Structures of neuroligin-1 and the neuroligin-1/neurexin-1 beta complex reveal specific protein-protein and protein-Ca2+ interactions. Neuron 56:992–1003
Ko J, Zhang C, Arac D, Boucard AA, Brunger AT, Südhof TC (2009) Neuroligin-1 performs neurexin-dependent and neurexin-independent functions in synapse validation. EMBO J 28:3244–3255
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Tsetsenis, T. (2017). Monitoring Synapses Via Trans-Synaptic GFP Complementation. In: Poulopoulos, A. (eds) Synapse Development. Methods in Molecular Biology, vol 1538. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6688-2_4
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DOI: https://doi.org/10.1007/978-1-4939-6688-2_4
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6686-8
Online ISBN: 978-1-4939-6688-2
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