Feng, Linqing Kwon, Osung Lee, Bokyoung Oh, Won Chan Kim, Jinhyun 2024-01-20T08:34:28Z 2024-01-20T08:34:28Z 2022-01-10 2014-10 1754-2189 https://pubs.kist.re.kr/handle/201004/126267 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. English NATURE PUBLISHING GROUP NERVOUS-SYSTEM NEURONS ELECTROPORATION EXPRESSION NEOCORTEX SYNAPSES SINGLE Using mammalian GFP reconstitution across synaptic partners (mGRASP) to map synaptic connectivity in the mouse brain Article 10.1038/nprot.2014.166 1 NATURE PROTOCOLS, v.9, no.10, pp.2425 - 2437 NATURE PROTOCOLS 9 10 2425 2437 scie scopus 000343227800011 2-s2.0-84923833833 Biochemical Research Methods Biochemistry & Molecular Biology Article NERVOUS-SYSTEM NEURONS ELECTROPORATION EXPRESSION NEOCORTEX SYNAPSES SINGLE