Kim, Jinhyun Zhao, Ting Petralia, Ronald S. Yu, Yang Peng, Hanchuan Myers, Eugene Magee, Jeffrey C. 2024-01-20T15:34:15Z 2024-01-20T15:34:15Z 2021-09-05 2012-01 1548-7091 https://pubs.kist.re.kr/handle/201004/129687 The GFP reconstitution across synaptic partners (GRASP) technique, based on functional complementation between two nonfluorescent GFP fragments, can be used to detect the location of synapses quickly, accurately and with high spatial resolution. The method has been previously applied in the nematode and the fruit fly but requires substantial modification for use in the mammalian brain. We developed mammalian GRASP (mGRASP) by optimizing transmembrane split-GFP carriers for mammalian synapses. Using in silico protein design, we engineered chimeric synaptic mGRASP fragments that were efficiently delivered to synaptic locations and reconstituted GFP fluorescence in vivo. Furthermore, by integrating molecular and cellular approaches with a computational strategy for the three-dimensional reconstruction of neurons, we applied mGRASP to both long-range circuits and local microcircuits in the mouse hippocampus and thalamocortical regions, analyzing synaptic distribution in single neurons and in dendritic compartments. English NATURE PUBLISHING GROUP EXPRESSION CIRCUIT BRAIN NEUREXINS SYNAPSES PROTEINS SINGLE ARCHITECTURE MEMBRANE mGRASP enables mapping mammalian synaptic connectivity with light microscopy Article 10.1038/NMETH.1784 1 NATURE METHODS, v.9, no.1, pp.96 - U139 NATURE METHODS 9 1 96 U139 scie scopus 000298667000037 2-s2.0-84856418376 Biochemical Research Methods Biochemistry & Molecular Biology Article EXPRESSION CIRCUIT BRAIN NEUREXINS SYNAPSES PROTEINS SINGLE ARCHITECTURE MEMBRANE