Bimodal Activation of Different Neuron Classes with the Spectrally Red-Shifted Channelrhodopsin Chimera C1V1 in Caenorhabditis elegansReport as inadecuate




Bimodal Activation of Different Neuron Classes with the Spectrally Red-Shifted Channelrhodopsin Chimera C1V1 in Caenorhabditis elegans - Download this document for free, or read online. Document in PDF available to download.

The C. elegans nervous system is particularly well suited for optogenetic analyses of circuit function: Essentially all connections have been mapped, and light can be directed at the neuron of interest in the freely moving, transparent animals, while behavior is observed. Thus, different nodes of a neuronal network can be probed for their role in controlling a particular behavior, using different optogenetic tools for photo-activation or –inhibition, which respond to different colors of light. As neurons may act in concert or in opposing ways to affect a behavior, one would further like to excite these neurons concomitantly, yet independent of each other. In addition to the blue-light activated Channelrhodopsin-2 ChR2, spectrally red-shifted ChR variants have been explored recently. Here, we establish the green-light activated ChR chimera C1V1 from Chlamydomonas and Volvox ChR1′s for use in C. elegans. We surveyed a number of red-shifted ChRs, and found that C1V1-ET-ET E122T; E162T works most reliable in C. elegans, with 540–580 nm excitation, which leaves ChR2 silent. However, as C1V1-ET-ET is very light sensitive, it still becomes activated when ChR2 is stimulated, even at 400 nm. Thus, we generated a highly efficient blue ChR2, the H134R; T159C double mutant ChR2-HR-TC. Both proteins can be used in the same animal, in different neurons, to independently control each cell type with light, enabling a further level of complexity in circuit analyses.



Author: Karen Erbguth, Matthias Prigge, Franziska Schneider, Peter Hegemann, Alexander Gottschalk

Source: http://plos.srce.hr/



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