Journal
NATURE METHODS
Volume 5, Issue 10, Pages 895-902Publisher
NATURE RESEARCH
DOI: 10.1038/NMETH.1252
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Funding
- Goethe University, Frankfurt
- Deutsche Forschungsgemeinschaft [SFB628, GO 1011/2-1]
- Cluster of Excellence Frankfurt, Macromolecular Complexes
- Canadian Institute of Health Research [MOP-79404, MOP-74530]
- US National Institutes of Health [R01 EY017241, P50 MH062196]
- Swartz Foundation
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We introduce optogenetic investigation of neurotransmission (OptIoN) for time-resolved and quantitative assessment of synaptic function via behavioral and electrophysiological analyses. We photo-triggered release of acetylcholine or gamma-aminobutyric acid at Caenorhabditis elegans neuromuscular junctions using targeted expression of Chlamydomonas reinhardtii Channelrhodopsin-2. In intact Channelrhodopsin-2 transgenic worms, photostimulation instantly induced body elongation (for gamma-aminobutyric acid) or contraction ( for acetylcholine), which we analyzed acutely, or during sustained activation with automated image analysis, to assess synaptic efficacy. In dissected worms, photostimulation evoked neurotransmitter-specific postsynaptic currents that could be triggered repeatedly and at various frequencies. Light-evoked behaviors and postsynaptic currents were significantly (P <= 0.05) altered in mutants with pre- or postsynaptic defects, although the behavioral phenotypes did not unambiguously report on synaptic function in all cases tested. OptIoN facilitates the analysis of neurotransmission with high temporal precision, in a neurotransmitter-selective manner, possibly allowing future investigation of synaptic plasticity in C. elegans.
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