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      Negative optokinetic afternystagmus in larval zebrafish demonstrates set-point adaptation

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          Abstract

          Motor learning is essential to maintain accurate behavioral responses. We used a larval zebrafish model to study ocular motor learning behaviors. During a sustained period of optokinetic stimulation in 5-day-old wild-type zebrafish larvae the slow-phase eye velocity decreased over time. Then interestingly, a long-lasting and robust negative optokinetic afternystagmus (OKAN) was evoked upon light extinction. The slow-phase velocity, the quick-phase frequency, and the decay time constant of the negative OKAN were dependent on the stimulus duration and the adaptation to the preceding optokinetic stimulation. Based on these results, we propose a sensory adaptation process during continued optokinetic stimulation, which, when the stimulus is removed, leads to a negative OKAN as the result of a changed retinal slip velocity set point, and thus, a sensorimotor memory. The pronounced negative OKAN in larval zebrafish not only provides a practical solution to the hitherto unsolved problems of observing negative OKAN, but also, and most importantly, can be readily applied as a powerful model for studying sensorimotor learning and memory in vertebrates.

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          The identification of genes with unique and essential functions in the development of the zebrafish, Danio rerio.

          In a large-scale screen, we isolated mutants displaying a specific visible phenotype in embryos or early larvae of the zebrafish, Danio rerio. Males were mutagenized with ethylnitrosourea (ENU) and F2 families of single pair matings between sibling F1 fish, heterozygous for a mutagenized genome, were raised. Egg lays were obtained from several crosses between F2 siblings, resulting in scoring of 3857 mutagenized genomes. F3 progeny were scored at the second, third and sixth day of development, using a stereomicroscope. In a subsequent screen, fixed embryos were analyzed for correct retinotectal projection. A total of 4264 mutants were identified. Two thirds of the mutants displaying rather general abnormalities were eventually discarded. We kept and characterized 1163 mutants. In complementation crosses performed between mutants with similar phenotypes, 894 mutants have been assigned to 372 genes. The average allele frequency is 2.4. We identified genes involved in early development, notochord, brain, spinal cord, somites, muscles, heart, circulation, blood, skin, fin, eye, otic vesicle, jaw and branchial arches, pigment pattern, pigment formation, gut, liver, motility and touch response. Our collection contains alleles of almost all previously described zebrafish mutants. From the allele frequencies and other considerations we estimate that the 372 genes defined by the mutants probably represent more than half of all genes that could have been discovered using the criteria of our screen. Here we give an overview of the spectrum of mutant phenotypes obtained, and discuss the limits and the potentials of a genetic saturation screen in the zebrafish.
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            Large-scale mutagenesis in the zebrafish: in search of genes controlling development in a vertebrate.

            In Drosophila melanogaster and Caenorhabditis elegans, the elucidation of developmental mechanisms has relied primarily on the systematic induction and isolation of mutations in genes with specific functions in development. Such an approach has not yet been possible in a vertebrate species, owing to the difficulty of analyzing and keeping a sufficiently high number of mutagenized lines of animals. We have developed the methods necessary to perform large-scale saturation screens for mutations affecting embryogenesis in the zebrafish, Danio (Brachydanio) rerio. Firstly, a new aquarium system was developed to raise and keep large numbers of strains of genetically different fish safely and with little maintenance care. Secondly, by placing adult male fish in water containing the chemical mutagen, ethylnitrosourea, we induced point mutations in premeiotic germ cells with a rate of one to three mutations per locus per 1,000 mutagenized haploid genomes. This rate, which is similar to the mutagenesis rates produced by ethylmethanesulfonate in Drosophila, was determined for alleles at four different pigmentation genes. Finally, in a pilot screen in which mutagenized fish were inbred for two generations and scored for embryonic mutants, we isolated 100 recessive mutations with phenotypes visible in the homozygous embryos. The high rate of induction and recovery of point mutations, in addition to an efficient aquarium system to house large numbers of mutagenized lines, means that it is now possible to perform saturation mutagenesis screens in a vertebrate, similar to those done in invertebrates.
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              Perspectives and problems in motor learning

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                Author and article information

                Contributors
                melody.yingyu.huang@gmail.com
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                13 December 2019
                13 December 2019
                2019
                : 9
                : 19039
                Affiliations
                [1 ]Department of Neurology, University Hospital Zurich, University of Zurich, Zurich, Switzerland
                [2 ]ISNI 0000 0004 1937 0650, GRID grid.7400.3, Neuroscience Center Zurich (ZNZ), , University of Zurich and ETH Zurich, ; Zurich, Switzerland
                [3 ]ISNI 0000 0004 1936 8649, GRID grid.14709.3b, Department of Biomedical Engineering, , McGill University, ; Montreal, Quebec Canada
                [4 ]ISNI 0000 0004 1936 7857, GRID grid.1002.3, Present Address: Monash Biomedicine Discovery Institute, , Monash University, ; Melbourne, Australia
                [5 ]ISNI 0000 0001 2171 9311, GRID grid.21107.35, Department of Biomedical Engineering, , The Johns Hopkins University, ; Baltimore, Maryland USA
                [6 ]ISNI 0000 0004 1936 8649, GRID grid.14709.3b, Department of Physiology, , McGill University, ; Montreal, Quebec Canada
                Author information
                http://orcid.org/0000-0002-6166-7461
                http://orcid.org/0000-0002-3032-452X
                Article
                55457
                10.1038/s41598-019-55457-4
                6910917
                31836778
                46c186ee-dc72-48d9-853d-13bf01973ca3
                © The Author(s) 2019

                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 August 2019
                : 27 November 2019
                Funding
                Funded by: This work was supported by the Oxford McGill ZNZ Partnership in the Neurosciences Pilot Project Funding (KEC, MJC and MYH), the Canadian Institutes of Health Research (MJC), the Research Grant for the Faculty of Medicine, UZH (MYH), the Dr. Dabbous Foundation (T-FL, AMF, DS and MYH), and the Betty and David Koetser Foundation for Brain Research (T-FL and MYH).
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                © The Author(s) 2019

                Uncategorized
                oculomotor system,reflexes
                Uncategorized
                oculomotor system, reflexes

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