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      The rise of photoresponsive protein technologies applications in vivo: a spotlight on zebrafish developmental and cell biology

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      Author(s): 1 , 2 , 3 , 4 , a , 1 , 2 , 3 , 4
      Publication date (Electronic):
      Journal: F1000Research
      Publisher: F1000Research
      Keywords: embryogenesis, CRE, optogenetic technology, fluorescent proteins

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          Abstract

          The zebrafish ( Danio rerio) is a powerful vertebrate model to study cellular and developmental processes in vivo. The optical clarity and their amenability to genetic manipulation make zebrafish a model of choice when it comes to applying optical techniques involving genetically encoded photoresponsive protein technologies. In recent years, a number of fluorescent protein and optogenetic technologies have emerged that allow new ways to visualize, quantify, and perturb developmental dynamics. Here, we explain the principles of these new tools and describe some of their representative applications in zebrafish.

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          The green fluorescent protein.

          R Tsien (1998)
          In just three years, the green fluorescent protein (GFP) from the jellyfish Aequorea victoria has vaulted from obscurity to become one of the most widely studied and exploited proteins in biochemistry and cell biology. Its amazing ability to generate a highly visible, efficiently emitting internal fluorophore is both intrinsically fascinating and tremendously valuable. High-resolution crystal structures of GFP offer unprecedented opportunities to understand and manipulate the relation between protein structure and spectroscopic function. GFP has become well established as a marker of gene expression and protein targeting in intact cells and organisms. Mutagenesis and engineering of GFP into chimeric proteins are opening new vistas in physiological indicators, biosensors, and photochemical memories.
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            Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy.

            Philipp Keller, Annette Schmidt, Joachim Wittbrodt … (2008)
            A long-standing goal of biology is to map the behavior of all cells during vertebrate embryogenesis. We developed digital scanned laser light sheet fluorescence microscopy and recorded nuclei localization and movement in entire wild-type and mutant zebrafish embryos over the first 24 hours of development. Multiview in vivo imaging at 1.5 billion voxels per minute provides "digital embryos," that is, comprehensive databases of cell positions, divisions, and migratory tracks. Our analysis of global cell division patterns reveals a maternally defined initial morphodynamic symmetry break, which identifies the embryonic body axis. We further derive a model of germ layer formation and show that the mesendoderm forms from one-third of the embryo's cells in a single event. Our digital embryos, with 55 million nucleus entries, are provided as a resource.
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              Sensitive red protein calcium indicators for imaging neural activity

              Hod Dana, Boaz Mohar, Yi Sun … (2016)
              Genetically encoded calcium indicators (GECIs) allow measurement of activity in large populations of neurons and in small neuronal compartments, over times of milliseconds to months. Although GFP-based GECIs are widely used for in vivo neurophysiology, GECIs with red-shifted excitation and emission spectra have advantages for in vivo imaging because of reduced scattering and absorption in tissue, and a consequent reduction in phototoxicity. However, current red GECIs are inferior to the state-of-the-art GFP-based GCaMP6 indicators for detecting and quantifying neural activity. Here we present improved red GECIs based on mRuby (jRCaMP1a, b) and mApple (jRGECO1a), with sensitivity comparable to GCaMP6. We characterized the performance of the new red GECIs in cultured neurons and in mouse, Drosophila, zebrafish and C. elegans in vivo. Red GECIs facilitate deep-tissue imaging, dual-color imaging together with GFP-based reporters, and the use of optogenetics in combination with calcium imaging. DOI: http://dx.doi.org/10.7554/eLife.12727.001
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                Author and article information

                Journal
                Journal ID (nlm-ta): F1000Res
                Journal ID (iso-abbrev): F1000Res
                Journal ID (pmc): F1000Research
                Title: F1000Research
                Publisher: F1000Research (London, UK )
                ISSN (Electronic): 2046-1402
                Publication date (Electronic): 11 April 2017
                Publication date Collection: 2017
                Volume: 6
                Electronic Location Identifier: F1000 Faculty Rev-459
                Affiliations
                [1 ]Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France
                [2 ]Centre National de la Recherche Scientifique UMR8104, Illkirch, France
                [3 ]Institut National de la Santé et de la Recherche Médicale, U964, Illkirch, France
                [4 ]Université de Strasbourg, Illkirch, France
                Author notes

                Competing interests: The authors declare that they have no competing interests.

                Author information
                http://orcid.org/0000-0002-8924-732X
                Article
                DOI: 10.12688/f1000research.10617.1
                PMC ID: 5389412
                PubMed ID: 28413613
                SO-VID: c52cb471-7afa-4966-abf8-00319eef44e7
                Copyright © Copyright: © 2017 Chow RWY and Vermot J
                License:

                This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                Date accepted : 6 April 2017
                Funding
                Funded by: Fondation pour la RechercheMédicale
                Award ID: DEQ20140329553
                Funded by: AgenceNationale de la Recherche
                Award ID: ANR-15-CE13-0015-01
                Award ID: ANR-12-ISV2-0001-01
                Award ID: ANR-10-LABX-0030-INRT
                Funded by: European Molecular Biology Organization Young Investigator Program
                Funded by: European Research Council
                Award ID: 682938
                Funded by: Investissementsd’Avenir
                Award ID: ANR-10-IDEX-0002-02
                JV and RC are supported by Fondation pour la Recherche Médicale (DEQ20140329553). JV is supported by the Agence Nationale de la Recherche (ANR) (ANR-15-CE13-0015-01 and ANR-12-ISV2-0001-01), the European Molecular Biology Organization Young Investigator Program, European Research Council consolidator grant Evalve (682938), and grant ANR-10-LABX-0030-INRT, a French State fund managed by the ANR under the frame program Investissements d’Avenir labeled ANR-10-IDEX-0002-02.
                Categories
                Subject: Review
                Subject: Articles
                Subject: Animal Genetics
                Subject: Behavioral Neuroscience
                Subject: Cardiovascular Physiology/Circulation
                Subject: Cell Signaling
                Subject: Chemical Biology of the Cell
                Subject: Developmental Evolution
                Subject: Developmental Molecular Mechanisms
                Subject: Genomics
                Subject: Morphogenesis & Cell Biology
                Subject: Neurodevelopment
                Subject: Neuronal & Glial Cell Biology
                Subject: Neuronal Signaling Mechanisms
                Subject: Pattern Formation
                Subject: Physiogenomics
                Subject: Protein Chemistry & Proteomics
                Subject: Stem Cells & Regeneration

                Keywords: embryogenesis,cre,optogenetic technology,fluorescent proteins
                Data availability:
                Keywords: embryogenesis, cre, optogenetic technology, fluorescent proteins

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