mGRASP enables mapping mammalian synaptic connectivity with light microscopy.
mGRASP enables mapping mammalian synaptic connectivity with light microscopy.
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DOI：
10.1038/nmeth.1784
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发表时间：
2011-12-04
期刊：
影响因子：
48
通讯作者：
Magee, Jeffrey C.
中科院分区：
文献类型：
--
作者：
Kim, Jinhyun;Zhao, Ting;Petralia, Ronald S.;Yu, Yang;Peng, Hanchuan;Myers, Eugene;Magee, Jeffrey C.
基于两个非荧光GFP片段之间的功能互补的GFP跨突触伴侣重建（GRASP）技术可用于快速、准确和高空间分辨率地检测突触的位置。该方法先前已应用于线虫和果蝇，但需要大量的修改用于哺乳动物的大脑。我们开发了哺乳动物GRASP（mGRASP）通过优化跨膜分裂GFP载体哺乳动物突触。使用计算机蛋白设计，我们设计了嵌合突触mGRASP片段，其被有效地递送到突触位置并在体内重建GFP荧光。此外，通过整合分子和细胞的方法与神经元的三维重建的计算策略，我们应用mGRASP的远程电路和本地微电路在小鼠海马和丘脑皮质区，分析突触分布在单个神经元和树突状细胞室。
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.
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