Abstract
Sparse manipulation of neuron excitability during free behavior is critical for identifying neural substrates of behavior. Genetic tools for precise neuronal manipulation exist in the fruit fly, Drosophila melanogaster, but behavioral tools are still lacking to identify potentially subtle phenotypes only detectible using high-throughput and high spatiotemporal resolution. We developed three assay components that can be used modularly to study natural and optogenetically induced behaviors. FlyGate automatically releases flies one at a time into an assay. FlyDetect tracks flies in real time, is robust to severe occlusions, and can be used to track appendages, such as the head. GlobeDisplay is a spherical projection system covering the fly's visual receptive field with a single projector. We demonstrate the utility of these components in an integrated system, FlyPEZ, by comprehensively modeling the input-output function for directional looming-evoked escape takeoffs and describing a millisecond-timescale phenotype from genetic silencing of a single visual projection neuron type.
Highlights
To understand how neural circuits coordinate movement and control complex behaviors, it is advantageous to perturb activity in sparse neuronal subsets while observing the behavioral consequences (Celik and Wernet, 2017; Robie et al, 2017; Wiegert et al, 2017), ideally during natural behaviors
We present a set of tools (FlyGate, FlyDetect, and GlobeDisplay) that provide solutions for three primary technical challenges to assaying natural behavior with simultaneous high resolution and high throughput: (1) how to rapidly and automatically isolate individual flies from aggregates without undue perturbation or visual stimulation, (2) how to track flies in three dimensions when they may be occluded in small chambers, and (3) how to target an individual for controlled sensory or optogenetic stimulation
We provide examples of other behaviors, including grooming, courtship, and proboscis extension, that we observe with the FlyPEZ
Summary
To understand how neural circuits coordinate movement and control complex behaviors, it is advantageous to perturb activity in sparse neuronal subsets while observing the behavioral consequences (Celik and Wernet, 2017; Robie et al, 2017; Wiegert et al, 2017), ideally during natural behaviors. Available neuroanatomical information and corresponding genetic reagents available in the fruit fly, Drosophila melanogaster, have refined the spatial (Jenett et al, 2012; Martın and Alcorta, 2017; Pfeiffer et al, 2010) and temporal (Bath et al, 2014; Inagaki et al, 2014; Klapoetke et al, 2014) targeting of such neuronal manipulations, offering an unprecedented opportunity to link neuronal activity in identified cell types to behavior. The quest to link sparse neuronal manipulation to free behavior phenotypes ideally requires a high-resolution, high-throughput assay in which social and sensory variables can be controlled
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