TY - JOUR
T1 - Rapid and reversible optogenetic silencing of synaptic transmission by clustering of synaptic vesicles
AU - Vettkötter, Dennis
AU - Schneider, Martin
AU - Goulden, Brady D.
AU - Dill, Holger
AU - Liewald, Jana
AU - Zeiler, Sandra
AU - Guldan, Julia
AU - Ateş, Yilmaz Arda
AU - Watanabe, Shigeki
AU - Gottschalk, Alexander
N1 - Funding Information:
We thank Chandra Tucker for providing CRY2 plasmids. We thank members of the Gottschalk lab for critical comments. We acknowledge the Caenorhabditis Genetic Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440), and the National Bioresource project, nematode C. elegans, for strains. We are indebted to Franziska Baumbach, Hans-Werner Müller, Barbara Janosi, Marion Basoglu, and Marius Seidenthal for expert technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG), SPP 1926, Project VIb (GO1011/12-2), GO1011/19-1, CRC1080 (Project B2), and by Goethe University Frankfurt to A.G.; S.W. is supported by National Institutes of Health (1DP2 NS111133-01 and 1R01 NS105810-01A1. S.W. is an Alfred P. Sloan fellow, McKnight Foundation Scholar, and Klingenstein and Simons Foundation scholar. B.D.G. was supported by the National Science Foundation Graduate research fellowship program (grant # DGE-2139757).
Funding Information:
We thank Chandra Tucker for providing CRY2 plasmids. We thank members of the Gottschalk lab for critical comments. We acknowledge the Caenorhabditis Genetic Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440), and the National Bioresource project, nematode C. elegans , for strains. We are indebted to Franziska Baumbach, Hans-Werner Müller, Barbara Janosi, Marion Basoglu, and Marius Seidenthal for expert technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG), SPP 1926, Project VIb (GO1011/12-2), GO1011/19-1, CRC1080 (Project B2), and by Goethe University Frankfurt to A.G.; S.W. is supported by National Institutes of Health (1DP2 NS111133-01 and 1R01 NS105810-01A1. S.W. is an Alfred P. Sloan fellow, McKnight Foundation Scholar, and Klingenstein and Simons Foundation scholar. B.D.G. was supported by the National Science Foundation Graduate research fellowship program (grant # DGE-2139757).
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - Acutely 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.
AB - Acutely 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.
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U2 - 10.1038/s41467-022-35324-z
DO - 10.1038/s41467-022-35324-z
M3 - Article
C2 - 36535932
AN - SCOPUS:85144302507
SN - 2041-1723
VL - 13
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 7827
ER -