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Published October 2018 | Submitted
Journal Article Open

Optogenetic manipulation of medullary neurons in the locust optic lobe

Abstract

The locust is a widely used animal model for studying sensory processing and its relation to behavior. Due to the lack of genomic information, genetic tools to manipulate neural circuits in locusts are not yet available. We examined whether Semliki Forest virus is suitable to mediate exogenous gene expression in neurons of the locust optic lobe. We subcloned a channelrhodopsin variant and the yellow fluorescent protein Venus into a Semliki Forest virus vector and injected the virus into the optic lobe of locusts (Schistocerca americana). Fluorescence was observed in all injected optic lobes. Most neurons that expressed the recombinant proteins were located in the first two neuropils of the optic lobe, the lamina and medulla. Extracellular recordings demonstrated that laser illumination increased the firing rate of medullary neurons expressing channelrhodopsin. The optogenetic activation of the medullary neurons also triggered excitatory postsynaptic potentials and firing of a postsynaptic, looming-sensitive neuron, the lobula giant movement detector. These results indicate that Semliki Forest virus is efficient at mediating transient exogenous gene expression and provides a tool to manipulate neural circuits in the locust nervous system and likely other insects.

Additional Information

© 2018 the American Physiological Society. Received 31 May 2018; Accepted 9 August 2018; Published online 9 October 2018; Published in print 1 October 2018. We thank Drs. Hiromu Yawo and Toru Ishizuka for providing pChopWR-Venus. We also thank Drs. Emma Victoria Jones and Keith Murai for providing the plasmids pENTR2B and pScaA7-RFA, and Dr. Alan L. Goldin for the pSFV-Helper2. This work was supported by a grant from the National Institute of Mental Health (MH-065339) and the National Science Foundation (IIS-1607518). No conflicts of interest, financial or otherwise, are declared by the authors. Author Contributions: H.W., M.U.E., J.R., M.L.R., and F.G. conceived and designed research; H.W., R.B.D., E.S., and J.R. performed experiments; H.W. and R.B.D. analyzed data; H.W., R.B.D., and F.G. interpreted results of experiments; H.W. and F.G. prepared figures; H.W. and F.G. drafted manuscript; H.W., R.B.D., M.U.E., and F.G. edited and revised manuscript; H.W., R.B.D., M.U.E., E.S., J.R., M.L.R., and F.G. approved final version of manuscript.

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August 22, 2023
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October 23, 2023