A method for recording behavior and multineuronal CNS activity from tethered insects flying in virtual space

We describe a low cost, novel virtual reality-based insect flight simulator that combines visual, olfactory and mechanosensory stimuli with multichannel neurophysiological recording techniques. Three-dimensional visual environments were created using customized modifications of a first person flight simulator computer game. Experiments could be performed in open-loop, where the flying insect's movement through the environment is ‘driven’ by the human operator, or in closed-loop where the movement of the environment is controlled by optically sensed movements of the insect's abdomen. During flight, we recorded multineuronal activity from the ventral nerve cord between the brain and thoracic ganglia. Results show that in open-loop conditions, induced turns of the environment evoked characteristic compensatory optomotor responses. Coordination of wing and body kinematics was similar to that observed in free flight. In closed-loop conditions, the insect was able to navigate through the simulated environment and produce flight tracks in response to presentation of pheromone that resemble those observed in free flight. We discuss the effectiveness of this preparation and its utility for addressing specific questions of insect flight as well as general questions in neuroethology.