Optogenetic Perturbation of Neural Activity with Laser Illumination in Semi-intact Drosophila Larvae in Motion
MyJove Corporation -- Journal of Visualized Experiments
DOI 10.3791/50513
Keyword(s)
  1. Neuroscience
  2. Issue 77
  3. Molecular Biology
  4. Neurobiology
  5. Developmental Biology
  6. Bioengineering
  7. Cellular Biology
  8. Motor Neurons
  9. Neurosciences
  10. Drosophila
  11. Optogenetics
  12. Channelrhodopsin-2
  13. Halorhodopsin
  14. laser
  15. confocal microscopy
  16. animal model
Abstract(s)

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.