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      Monitoring of Single-Cell Responses in the Optic Tectum of Adult Zebrafish with Dextran-Coupled Calcium Dyes Delivered via Local Electroporation

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      PLoS ONE
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          Abstract

          The zebrafish ( Danio rerio) has become one of the major animal models for in vivo examination of sensory and neuronal computation. Similar to Xenopus tadpoles neural activity in the optic tectum, the major region controlling visually guided behavior, can be examined in zebrafish larvae by optical imaging. Prerequisites of these approaches are usually the transparency of larvae up to a certain age and the use of two-photon microscopy. This principle of fluorescence excitation was necessary to suppress crosstalk between signals from individual neurons, which is a critical issue when using membrane-permeant dyes. This makes the equipment to study neuronal processing costly and limits the approach to the study of larvae. Thus there is lack of knowledge about the properties of neurons in the optic tectum of adult animals. We established a procedure to circumvent these problems, enabling in vivo calcium imaging in the optic tectum of adult zebrafish. Following local application of dextran-coupled dyes single-neuron activity of adult zebrafish can be monitored with conventional widefield microscopy, because dye labeling remains restricted to tens of neurons or less. Among the neurons characterized with our technique we found neurons that were selective for a certain pattern orientation as well as neurons that responded in a direction-selective way to visual motion. These findings are consistent with previous studies and indicate that the functional integrity of neuronal circuits in the optic tectum of adult zebrafish is preserved with our staining technique. Overall, our protocol for in vivo calcium imaging provides a useful approach to monitor visual responses of individual neurons in the optic tectum of adult zebrafish even when only widefield microscopy is available. This approach will help to obtain valuable insight into the principles of visual computation in adult vertebrates and thus complement previous work on developing visual circuits.

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          Author and article information

          Contributors
          Role: Editor
          Journal
          PLoS One
          PLoS ONE
          plos
          plosone
          PLoS ONE
          Public Library of Science (San Francisco, USA )
          1932-6203
          2013
          7 May 2013
          : 8
          : 5
          : e62846
          Affiliations
          [1 ]AG Active Sensing and Center of Excellence ‘Cognitive Interaction Technology’, Bielefeld University, Bielefeld, Germany
          [2 ]Department of Neurobiology, Bielefeld University, Bielefeld, Germany
          Institut Curie, France
          Author notes

          Competing Interests: The authors have declared that no competing interests exist.

          Conceived and designed the experiments: VK JE RK. Performed the experiments: VK. Analyzed the data: VK JE RK. Wrote the paper: VK JE RK.

          Article
          PONE-D-12-39881
          10.1371/journal.pone.0062846
          3647071
          23667529
          1037cb80-a600-434c-a5f3-53b17c41148b
          Copyright @ 2013

          This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

          History
          : 17 December 2012
          : 26 March 2013
          Page count
          Pages: 10
          Funding
          VK was funded by a scholarship of the “Deutsche Forschungsgemeinschaft (DFG) - Excellence Cluster 277: Cognitive Interaction Technology (CITEC),” http://www.dfg.de/en/research_funding/programmes/list/projectdetails/index.jsp?id=39113330. The authors acknowledge support for the article processing charge by the Deutsche Forschungsgemeinschaft and the Open Access Publication Funds of Bielefeld University Library. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
          Categories
          Research Article
          Biology
          Histology
          Model Organisms
          Animal Models
          Zebrafish
          Neuroscience
          Neuroimaging
          Calcium Imaging
          Sensory Systems
          Visual System
          Cellular Neuroscience
          Neurophysiology
          Engineering
          Signal Processing
          Image Processing

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          Uncategorized

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