Abstract
AbstractAcutely silencing specific neurons informs about their functional roles in circuits and behavior. Existing optogenetic silencers include ion pumps, channels, metabotropic receptors, and tools that damage the neurotransmitter release machinery. While the former hyperpolarize the cell, alter ionic gradients or cellular biochemistry, the latter allow only slow recovery, requiring de novo synthesis. Thus, tools combining fast activation and reversibility are needed. Here, we use light-evoked homo-oligomerization of cryptochrome CRY2 to silence synaptic transmission, by clustering synaptic vesicles (SVs). We benchmark this tool, optoSynC, in Caenorhabditis elegans, zebrafish, and murine hippocampal neurons. optoSynC clusters SVs, observable by electron microscopy. Locomotion silencing occurs with tauon ~7.2 s and recovers with tauoff ~6.5 min after light-off. optoSynC can inhibit exocytosis for several hours, at very low light intensities, does not affect ion currents, biochemistry or synaptic proteins, and may further allow manipulating different SV pools and the transfer of SVs between them.
Funder
Deutsche Forschungsgemeinschaft
Publisher
Springer Science and Business Media LLC
Subject
General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry,Multidisciplinary
Reference83 articles.
1. Calahorro, F. & Izquierdo, P. G. The presynaptic machinery at the synapse of C. elegans. Invert. Neurosci. 18, 4 (2018).
2. Denker, A. & Rizzoli, S. O. Synaptic vesicle pools: an update. Front. Synaptic Neurosci. 2, 135 (2010).
3. Brunger, A. T., Choi, U. B., Lai, Y., Leitz, J. & Zhou, Q. Molecular Mechanisms of Fast Neurotransmitter Release. Annu. Rev. Biophys. 47, 469–497 (2018).
4. Sudhof, T. C. Neurotransmitter release: the last millisecond in the life of a synaptic vesicle. Neuron 80, 675–690 (2013).
5. Jung, N. & Haucke, V. Clathrin-mediated endocytosis at synapses. Traffic 8, 1129–1136 (2007).
Cited by
8 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献