Journal
NEURON
Volume 79, Issue 2, Pages 241-253Publisher
CELL PRESS
DOI: 10.1016/j.neuron.2013.05.022
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Funding
- Foundation of Research, Science and Technology New Zealand
- Ruth L. Kirschstein National Research Service Awards (NIH NINDS) [NS067891]
- Instituts de Recherche en Sante du Canada
- National Institutes of Health [MH091119, NS035546, NS027177]
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Optogenetic techniques provide effective ways of manipulating the functions of selected neurons with light. In the current study, we engineered an optogenetic technique that directly inhibits neurotransmitter release. We used a genetically encoded singlet oxygen generator, miniSOG, to conduct chromophore assisted light inactivation (CALI) of synaptic proteins. Fusions of miniSOG to VAMP2 and synaptophysin enabled disruption of presynaptic vesicular release upon illumination with blue light. In cultured neurons and hippocampal organotypic slices, synaptic release was reduced up to 100%. Such inhibition lasted >1 hr and had minimal effects on membrane electrical properties. When miniSOG-VAMP2 was expressed panneuronally in Caenorhabditis elegans, movement of the worms was reduced after illumination, and paralysis was often observed. The movement of the worms recovered overnight. We name this technique Inhibition of Synapses with CALI (InSynC). InSynC is a powerful way to silence genetically specified synapses with light in a spatially and temporally precise manner.
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