title: Using mammalian GFP reconstitution across synaptic partners (mGRASP) to map synaptic connectivity in the mouse brain.
abstract: Many types of questions in neuroscience require the detection and mapping of synapses in the complex mammalian brain. A tool, mammalian GFP reconstitution across synaptic partners (mGRASP), offers a relatively easy, quick and economical approach to this technically challenging task. Here we describe in step-by-step detail the protocols for virus production, gene delivery, brain specimen preparation, fluorescence imaging and image analysis, calibrated substantially and specifically to make mGRASP-assisted circuit mapping (mGRASPing) practical in the mouse brain. The protocol includes troubleshooting suggestions and solutions to common problems. The mGRASP method is suitable for mapping mammalian synaptic connectivity at multiple scales: microscale for synapse-by-synapse or neuron-by-neuron analysis, and mesoscale for revealing local and long-range circuits. The entire protocol takes 5-6 weeks, including time for incubation and virus expression.
authors.affiliation: 1] Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul, Korea. [2].
authors.forename: Linqing
authors.initials: L
authors.lastname: Feng
authors.affiliation: 1] Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul, Korea. [2] Neuroscience program, University of Science and Technology, Daejeon, Korea. [3].
authors.forename: Osung
authors.initials: O
authors.lastname: Kwon
authors.affiliation: 1] Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul, Korea. [2] Neuroscience program, University of Science and Technology, Daejeon, Korea.
authors.forename: Bokyoung
authors.initials: B
authors.lastname: Lee
authors.affiliation: Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul, Korea.
authors.forename: Won Chan
authors.initials: WC
authors.lastname: Oh
authors.affiliation: 1] Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul, Korea. [2] Neuroscience program, University of Science and Technology, Daejeon, Korea.
authors.forename: Jinhyun
authors.initials: J
authors.lastname: Kim
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001931:Brain Mapping / Q000295:instrumentation / Q000379:methods*; D002138:Calibration; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008322:Mammals; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013569:Synapses / Q000378:metabolism*; D014162:Transfection
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins