Transgenic Strategy for Identifying Synaptic Connections in Mice by Fluorescence Complementation (GRASP)

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Transgenic Strategy for Identifying Synaptic Connections in Mice by Fluorescence Complementation (GRASP)

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dc.contributor.author Yamagata, Masahito
dc.contributor.author Sanes, Joshua R.
dc.date.accessioned 2012-04-03T17:23:53Z
dc.date.issued 2012
dc.identifier.citation Yamagata, Masahito, and Joshua R. Sanes. 2012. Transgenic strategy for identifying synaptic connections in mice by fluorescence complementation (GRASP). Frontiers in Molecular Neuroscience 5: 18. en_US
dc.identifier.issn 1662-5099 en_US
dc.identifier.uri http://nrs.harvard.edu/urn-3:HUL.InstRepos:8497447
dc.description.abstract In the “GFP reconstitution across synaptic partners” (GRASP) method, non-fluorescent fragments of GFP are expressed in two different neurons; the fragments self-assemble at synapses between the two to form a fluorophore. GRASP has proven useful for light microscopic identification of synapses in two invertebrate species, \(Caenorhabditis\) \(elegans\) and \(Drosophila\) \(melanogaster\), but has not yet been applied to vertebrates. Here, we describe GRASP constructs that function in mammalian cells and implement a transgenic strategy in which a Cre-dependent gene switch leads to expression of the two fragments in mutually exclusive neuronal subsets in mice. Using a transgenic line that expresses Cre selectively in rod photoreceptors, we demonstrate labeling of synapses in the outer plexiform layer of the retina. Labeling is specific, in that synapses made by rods remain labeled for at least 6 months whereas nearby synapses made by intercalated cone photoreceptors on many of the same interneurons remain unlabeled. We also generated antisera that label reconstituted GFP but neither fragment in order to amplify the GRASP signal and thereby increase the sensitivity of the method. en_US
dc.description.sponsorship Molecular and Cellular Biology en_US
dc.language.iso en_US en_US
dc.publisher Frontiers Research Foundation en_US
dc.relation.isversionof doi:10.3389/fnmol.2012.00018 en_US
dc.relation.hasversion http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3280602/pdf/ en_US
dash.license LAA
dc.subject GRASP en_US
dc.subject GFP en_US
dc.subject synapse en_US
dc.subject retina en_US
dc.subject photoreceptor en_US
dc.subject rod en_US
dc.subject neuroligin en_US
dc.subject neurexin en_US
dc.title Transgenic Strategy for Identifying Synaptic Connections in Mice by Fluorescence Complementation (GRASP) en_US
dc.type Journal Article en_US
dc.description.version Version of Record en_US
dc.relation.journal Frontiers in Molecular Neuroscience en_US
dash.depositing.author Sanes, Joshua R.
dc.date.available 2012-04-03T17:23:53Z

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  • FAS Scholarly Articles [7495]
    Peer reviewed scholarly articles from the Faculty of Arts and Sciences of Harvard University

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