Rapid and reversible optogenetic silencing of synaptic transmission by clustering of synaptic vesicles

Silencing neurons acutely and specifically informs about their functional roles in circuits and behavior. Existing optogenetic silencers include ion pumps or channels, and tools that damage the neurotransmitter release machinery. While the former hyperpolarize the cell and can alter ionic gradients, the latter allow only slow recovery, requiring de novo synthesis. Thus, there is a need for other strategies combining fast activation and reversibility. Here, we use light-evoked homo-oligomerization of cryptochrome CRY2 to silence synaptic transmission, by clustering synaptic vesicles (SVs). We benchmark this tool, optoSynC, by electrophysiology and locomotion in Caenorhabditis elegans . optoSynC clusters SVs within 25 s, causing approximation, observable by electron microscopy. Locomotion silencing is rapid (tau on ∼15 s) and recovers quickly (tau off ∼10 min) after light-off. Further, optoSynC can inhibit exocytosis for several hours, at very low light intensities. optoSynC is a highly efficient, ‘non-ionic’ optogenetic silencer that may further allow to manipulate different SV pools.