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      Deep learning-based behavioral analysis reaches human accuracy and is capable of outperforming commercial solutions

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          Abstract

          To study brain function, preclinical research heavily relies on animal monitoring and the subsequent analyses of behavior. Commercial platforms have enabled semi high-throughput behavioral analyses by automating animal tracking, yet they poorly recognize ethologically relevant behaviors and lack the flexibility to be employed in variable testing environments. Critical advances based on deep-learning and machine vision over the last couple of years now enable markerless tracking of individual body parts of freely moving rodents with high precision. Here, we compare the performance of commercially available platforms (EthoVision XT14, Noldus; TSE Multi-Conditioning System, TSE Systems) to cross-verified human annotation. We provide a set of videos—carefully annotated by several human raters—of three widely used behavioral tests (open field test, elevated plus maze, forced swim test). Using these data, we then deployed the pose estimation software DeepLabCut to extract skeletal mouse representations. Using simple post-analyses, we were able to track animals based on their skeletal representation in a range of classic behavioral tests at similar or greater accuracy than commercial behavioral tracking systems. We then developed supervised machine learning classifiers that integrate the skeletal representation with the manual annotations. This new combined approach allows us to score ethologically relevant behaviors with similar accuracy to humans, the current gold standard, while outperforming commercial solutions. Finally, we show that the resulting machine learning approach eliminates variation both within and between human annotators. In summary, our approach helps to improve the quality and accuracy of behavioral data, while outperforming commercial systems at a fraction of the cost.

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          DeepLabCut: markerless pose estimation of user-defined body parts with deep learning

          Quantifying behavior is crucial for many applications in neuroscience. Videography provides easy methods for the observation and recording of animal behavior in diverse settings, yet extracting particular aspects of a behavior for further analysis can be highly time consuming. In motor control studies, humans or other animals are often marked with reflective markers to assist with computer-based tracking, but markers are intrusive, and the number and location of the markers must be determined a priori. Here we present an efficient method for markerless pose estimation based on transfer learning with deep neural networks that achieves excellent results with minimal training data. We demonstrate the versatility of this framework by tracking various body parts in multiple species across a broad collection of behaviors. Remarkably, even when only a small number of frames are labeled (~200), the algorithm achieves excellent tracking performance on test frames that is comparable to human accuracy.
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            The use of the elevated plus maze as an assay of anxiety-related behavior in rodents.

            The elevated plus maze is a widely used behavioral assay for rodents and it has been validated to assess the anti-anxiety effects of pharmacological agents and steroid hormones, and to define brain regions and mechanisms underlying anxiety-related behavior. Briefly, rats or mice are placed at the junction of the four arms of the maze, facing an open arm, and entries/duration in each arm are recorded by a video-tracking system and observer simultaneously for 5 min. Other ethological parameters (i.e., rears, head dips and stretched-attend postures) can also be observed. An increase in open arm activity (duration and/or entries) reflects anti-anxiety behavior. In our laboratory, rats or mice are exposed to the plus maze on one occasion; thus, results can be obtained in 5 min per rodent.
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              Behavioural despair in rats: a new model sensitive to antidepressant treatments.

              Rats when forced to swim in a cylinder from which they cannot escape will, after an initial period of vigorous activity, adopt a characteristic immobile posture which can be readily identified. Immobility was reduced by various clinically effective antidepressant drugs at doses which otherwise decreased spontaneous motor activity in an open field. Antidepressants could thus be distinguished from psychostimulants which decreased immobility at doses which increased general activity. Anxiolytic compounds did not affect immobility whereas major tranquilisers enhanced it. Immobility was also reduced by electroconvulsive shock, REM sleep deprivation and "enrichment" of the environment. It was concluded that immobility reflects a state of lowered mood in the rat which is selectively sensitive to antidepressant treatments. Positive findings with atypical antidepressant drugs such as iprindole and mianserin suggest that the method may be capable of discovering new antidepressants hitherto undetectable with classical pharmacological tests.
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                Author and article information

                Contributors
                bohacekj@ethz.ch
                Journal
                Neuropsychopharmacology
                Neuropsychopharmacology
                Neuropsychopharmacology
                Springer International Publishing (Cham )
                0893-133X
                1740-634X
                25 July 2020
                25 July 2020
                October 2020
                : 45
                : 11
                : 1942-1952
                Affiliations
                [1 ]GRID grid.5801.c, ISNI 0000 0001 2156 2780, Laboratory of Molecular and Behavioral Neuroscience, Institute for Neuroscience, Department of Health Sciences and Technology, , ETH Zurich, ; Zurich, Switzerland
                [2 ]GRID grid.7400.3, ISNI 0000 0004 1937 0650, Neuroscience Center Zurich, , ETH Zurich and University of Zurich, ; Zurich, Switzerland
                [3 ]GRID grid.5801.c, ISNI 0000 0001 2156 2780, Neural Control of Movement Lab, Department of Health Sciences and Technology, , ETH Zurich, ; Zurich, Switzerland
                [4 ]GRID grid.7400.3, ISNI 0000 0004 1937 0650, Experimental Imaging and Neuroenergetics, Institute of Pharmacology and Toxicology, , University of Zurich, ; Zurich, Switzerland
                [5 ]GRID grid.7400.3, ISNI 0000 0004 1937 0650, Institute of Neuroinformatics, , University of Zurich and ETH Zurich, ; Zurich, Switzerland
                [6 ]GRID grid.5801.c, ISNI 0000 0001 2156 2780, Department of Information Technology and Electrical Engineering, , ETH Zurich, ; Zurich, Switzerland
                Author information
                http://orcid.org/0000-0002-8442-653X
                Article
                776
                10.1038/s41386-020-0776-y
                7608249
                32711402
                91d75e81-33bc-4a33-90fe-484a8066a0ec
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 1 May 2020
                : 13 July 2020
                : 15 July 2020
                Funding
                Funded by: ETH Zurich, ETH Project Grant ETH-20 19-1, the SNSF Grant CRSII5-173721, Swiss Data Science Center C17-18, Neuroscience Center Zurich Project Grants Oxford/McGill/Zurich Partnership.
                Funded by: ETH Zurich, ETH Project Grant ETH-20 19-1, the SNSF Grant 310030_172889/1, Forschungskredit of the University of Zurich FK-15-035, Vontobel-Foundation, Novartis Foundation for Medical Biological Research, EMDO-Foundation, Olga Mayenfisch Foundation, Betty and David Koetser Foundation for Brain Research, Neuroscience Center Zurich Project Grants Oxford/McGill/Zurich Partnership
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                © American College of Neuropsychopharmacology 2020

                Pharmacology & Pharmaceutical medicine
                anxiety,behavioural methods
                Pharmacology & Pharmaceutical medicine
                anxiety, behavioural methods

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