Optogenetic Perturbation of Neural Activity with Laser Illumination in Semi-intact Drosophila Larvae in Motion |
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Authors: | Teruyuki Matsunaga Akira Fushiki Akinao Nose Hiroshi Kohsaka |
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Affiliation: | 1.Department of Complexity Science and Engineering, Graduate School of Frontier Sciences, The University of Tokyo;2.Department of Physics, Graduate School of Science, The University of Tokyo |
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Abstract: | Drosophila larval locomotion is a splendid model system in developmental and physiological neuroscience, by virtue of the genetic accessibility of the underlying neuronal components in the circuits1-6. Application of optogenetics7,8 in the larval neural circuit allows us to manipulate neuronal activity in spatially and temporally patterned ways9-13. Typically, specimens are broadly illuminated with a mercury lamp or LED, so specificity of the target neurons is controlled by binary gene expression systems such as the Gal4-UAS system14,15. In this work, to improve the spatial resolution to "sub-genetic resolution", we locally illuminated a subset of neurons in the ventral nerve cord using lasers implemented in a conventional confocal microscope. While monitoring the motion of the body wall of the semi-intact larvae, we interactively activated or inhibited neural activity with channelrhodopsin16,17 or halorhodopsin18-20, respectively. By spatially and temporally restricted illumination of the neural tissue, we can manipulate the activity of specific neurons in the circuit at a specific phase of behavior. This method is useful for studying the relationship between the activities of a local neural assembly in the ventral nerve cord and the spatiotemporal pattern of motor output. |
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Keywords: | Neuroscience Issue 77 Molecular Biology Neurobiology Developmental Biology Bioengineering Cellular Biology Motor Neurons Neurosciences Drosophila Optogenetics Channelrhodopsin-2 Halorhodopsin laser confocal microscopy animal model |
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