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Abstract

Background: Drosophila shows a range of visually guided memory and learning behaviors, including place learning. Investigating the dynamics of neural circuits underlying such behaviors requires learning assays in tethered animals, compatible with in vivo imaging experiments.
New Method: Here, we introduce an assay for place learning for tethered walking flies. A cylindrical arena is rotated and translated in real time around the fly in concert with the rotational and translational walking activity measured with an air supported ball, resulting in a mechanical virtual reality (VR).Navigation together with heat-based operant conditioning allows flies to learn the location of a cool spot with respect to a visual landmark. Flies optimize the time and distance required to find the cool spot over a similar number of trials as observed in assays with freely moving flies. Additionally, a fraction of flies remembers the location of the cool spot also after the conditioning heat is removed.
Comparison with Existing Methods: Learning tasks have been implemented in tethered flying as well as walking flies. Mechanically translating and rotating an arena in concert with the fly’s walking activity enables navigation in a three dimensional environment.
Conclusion:In the developed mechanical VR flies can learn to remember the location of a cool place within an otherwise hot environment with respect to a visual landmark. Implementing place learning in a tethered walking configuration is a precondition for investigating the underlying circuit dynamics using functional imaging.