Abstract
As zebrafish emerge as a species of choice for the investigation of biological processes, a number of experimental protocols are being developed to study their social behaviour. While live stimuli may elicit varying response in focal subjects owing to idiosyncrasies, tiredness and circadian rhythms, video stimuli suffer from the absence of physical input and rely only on two-dimensional projections. Robotics has been recently proposed as an alternative approach to generate physical, customizable, effective and consistent stimuli for behavioural phenotyping. Here, we contribute to this field of investigation through a novel four-degree-of-freedom robotics-based platform to manoeuvre a biologically inspired three-dimensionally printed replica. The platform enables three-dimensional motions as well as body oscillations to mimic zebrafish locomotion. In a series of experiments, we demonstrate the differential role of the visual stimuli associated with the biologically inspired replica and its three-dimensional motion. Three-dimensional tracking and information-theoretic tools are complemented to quantify the interaction between zebrafish and the robotic stimulus. Live subjects displayed a robust attraction towards the moving replica, and such attraction was lost when controlling for its visual appearance or motion. This effort is expected to aid zebrafish behavioural phenotyping, by offering a novel approach to generate physical stimuli moving in three dimensions.
Highlights
In recent years, zebrafish (Danio rerio) has been extensively used in laboratory experiments for the investigation of a broad spectrum of functional and dysfunctional processes, such as neurological disorders [1], vertebrate embryonic development [2,3], preclinical toxicological assessment [4,5] and pharmacological assays [6].2016 The Authors
We studied the behavioural response of zebrafish to a biologically inspired three-dimensionally printed replica, which was fabricated and painted to mimic zebrafish morphology
The replica was manoeuvred by a novel four-degree-of-freedom robotic platform, which was designed to move the replica along sampled three-dimensional trajectories of live subjects and, independently, control its body oscillation
Summary
Zebrafish (Danio rerio) has been extensively used in laboratory experiments for the investigation of a broad spectrum of functional and dysfunctional processes, such as neurological disorders [1], vertebrate embryonic development [2,3], preclinical toxicological assessment [4,5] and pharmacological assays [6].2016 The Authors. A popular paradigm consists of studying the spatial preference of zebrafish in binary choice tests, using live [17,19,20] or video stimuli [21,22,23] In these experiments, fish behaviour is often scored in terms of time spent in the proximity of the stimuli [24], the average distance between fish and stimuli [25] or the number of entries in specific tank sections [26]. Fish behaviour is often scored in terms of time spent in the proximity of the stimuli [24], the average distance between fish and stimuli [25] or the number of entries in specific tank sections [26] These quantities are typically studied by recording zebrafish swimming patterns in two dimensions and analysing the acquired videos manually or through automated software [8]
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