title: Optogenetic perturbation of neural activity with laser illumination in semi-intact drosophila larvae in motion.
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 circuits(1-6). Application of optogenetics(7,8) in the larval neural circuit allows us to manipulate neuronal activity in spatially and temporally patterned ways(9-13). 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 system(14,15). 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 channelrhodopsin(16,17) or halorhodopsin(18-20), 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.
authors.affiliation: Department of Complexity Science and Engineering, Graduate School of Frontier Sciences, The University of Tokyo.
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Matsunaga
authors.forename: Akira
authors.initials: A
authors.lastname: Fushiki
authors.forename: Akinao
authors.initials: A
authors.lastname: Nose
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kohsaka
mesh_terms: D000818:Animals; D004330:Drosophila / Q000502:physiology*; D007814:Larva; D007834:Lasers; D008124:Locomotion / Q000502:physiology*; D018613:Microscopy, Confocal; D009046:Motor Neurons / Q000502:physiology*; D010528:Peristalsis / Q000502:physiology