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      A multi-animal tracker for studying complex behaviors

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          Abstract

          Background

          Animals exhibit astonishingly complex behaviors. Studying the subtle features of these behaviors requires quantitative, high-throughput, and accurate systems that can cope with the often rich perplexing data.

          Results

          Here, we present a Multi-Animal Tracker (MAT) that provides a user-friendly, end-to-end solution for imaging, tracking, and analyzing complex behaviors of multiple animals simultaneously. At the core of the tracker is a machine learning algorithm that provides immense flexibility to image various animals (e.g., worms, flies, zebrafish, etc.) under different experimental setups and conditions. Focusing on C. elegans worms, we demonstrate the vast advantages of using this MAT in studying complex behaviors. Beginning with chemotaxis, we show that approximately 100 animals can be tracked simultaneously, providing rich behavioral data. Interestingly, we reveal that worms’ directional changes are biased, rather than random – a strategy that significantly enhances chemotaxis performance. Next, we show that worms can integrate environmental information and that directional changes mediate the enhanced chemotaxis towards richer environments. Finally, offering high-throughput and accurate tracking, we show that the system is highly suitable for longitudinal studies of aging- and proteotoxicity-associated locomotion deficits, enabling large-scale drug and genetic screens.

          Conclusions

          Together, our tracker provides a powerful and simple system to study complex behaviors in a quantitative, high-throughput, and accurate manner.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12915-017-0363-9) contains supplementary material, which is available to authorized users.

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          Most cited references35

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          Stochastic and genetic factors influence tissue-specific decline in ageing C. elegans.

          The nematode Caenorhabditis elegans is an important model for studying the genetics of ageing, with over 50 life-extension mutations known so far. However, little is known about the pathobiology of ageing in this species, limiting attempts to connect genotype with senescent phenotype. Using ultrastructural analysis and visualization of specific cell types with green fluorescent protein, we examined cell integrity in different tissues as the animal ages. We report remarkable preservation of the nervous system, even in advanced old age, in contrast to a gradual, progressive deterioration of muscle, resembling human sarcopenia. The age-1(hx546) mutation, which extends lifespan by 60-100%, delayed some, but not all, cellular biomarkers of ageing. Strikingly, we found strong evidence that stochastic as well as genetic factors are significant in C. elegans ageing, with extensive variability both among same-age animals and between cells of the same type within individuals.
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            High-throughput Ethomics in Large Groups of Drosophila

            We present a camera-based method for automatically quantifying the individual and social behaviors of fruit flies, Drosophila melanogaster, interacting within a planar arena. Our system includes machine vision algorithms that accurately track many individuals without swapping identities and classification algorithms that detect behaviors. The data may be represented as an ethogram that plots the time course of behaviors exhibited by each fly, or as a vector that concisely captures the statistical properties of all behaviors displayed within a given period. We found that behavioral differences between individuals are consistent over time and are sufficient to accurately predict gender and genotype. In addition, we show that the relative positions of flies during social interactions vary according to gender, genotype, and social environment. We expect that our software, which permits high-throughput screening, will complement existing molecular methods available in Drosophila, facilitating new investigations into the genetic and cellular basis of behavior.
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              The threshold for polyglutamine-expansion protein aggregation and cellular toxicity is dynamic and influenced by aging in Caenorhabditis elegans.

              Studies of the mutant gene in Huntington's disease, and for eight related neurodegenerative disorders, have identified polyglutamine (polyQ) expansions as a basis for cellular toxicity. This finding has led to a disease hypothesis that protein aggregation and cellular dysfunction can occur at a threshold of approximately 40 glutamine residues. Here, we test this hypothesis by expression of fluorescently tagged polyQ proteins (Q29, Q33, Q35, Q40, and Q44) in the body wall muscle cells of Caenorhabditis elegans and show that young adults exhibit a sharp boundary at 35-40 glutamines associated with the appearance of protein aggregates and loss of motility. Surprisingly, genetically identical animals expressing near-threshold polyQ repeats exhibited a high degree of variation in the appearance of protein aggregates and cellular toxicity that was dependent on repeat length and exacerbated during aging. The role of genetically determined aging pathways in the progression of age-dependent polyQ-mediated aggregation and cellular toxicity was tested by expressing Q82 in the background of age-1 mutant animals that exhibit an extended lifespan. We observed a dramatic delay of polyQ toxicity and appearance of protein aggregates. These data provide experimental support for the threshold hypothesis of polyQ-mediated toxicity in an experimental organism and emphasize the importance of the threshold as a point at which genetic modifiers and aging influence biochemical environment and protein homeostasis in the cell.
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                Author and article information

                Contributors
                alonzas@mail.huji.ac.il
                Journal
                BMC Biol
                BMC Biol
                BMC Biology
                BioMed Central (London )
                1741-7007
                6 April 2017
                6 April 2017
                2017
                : 15
                : 29
                Affiliations
                [1 ]GRID grid.9619.7, Department of Genetics, The Silberman Institute of Life Science, , Edmond J. Safra Campus, The Hebrew University of Jerusalem, ; Jerusalem, 91904 Israel
                [2 ]GRID grid.9619.7, School of Computer Science and Engineering, , Hebrew University, ; Jerusalem, Israel
                [3 ]GRID grid.9619.7, Biochemistry and Molecular Biology, The Institute for Medical Research Israel – Canada (IMRIC), School of Medicine, , The Hebrew University of Jerusalem, ; Jerusalem, 91120 Israel
                Author information
                http://orcid.org/0000-0002-7871-4420
                Article
                363
                10.1186/s12915-017-0363-9
                5383998
                28385158
                6255ae4c-fcf6-4d86-8fd8-f95f7e60b338
                © Zaslaver et al. 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 9 February 2017
                : 6 March 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100000781, European Research Council;
                Award ID: 336803
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000965, American Federation for Aging Research;
                Categories
                Methodology Article
                Custom metadata
                © The Author(s) 2017

                Life sciences
                multi-animal tracking,image analyses,chemotaxis,c. elegans,locomotion
                Life sciences
                multi-animal tracking, image analyses, chemotaxis, c. elegans, locomotion

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