Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation.
Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation.
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DOI：
10.1038/ncomms10024
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发表时间：
2015-12-04
影响因子：
16.6
通讯作者：
Gallio M
中科院分区：
文献类型：
--
作者：
Macpherson LJ;Zaharieva EE;Kearney PJ;Alpert MH;Lin TY;Turan Z;Lee CH;Gallio M
确定神经回路内各个连接的活动模式可以深入了解大脑功能的计算过程。在这里，我们开发了通过果蝇跨突触荧光互补来标记活性突触的新策略。首先，我们证明突触短蛋白-GRASP嵌合体可作为体内突触的强大活性依赖性标记物。接下来，我们创建青色和黄色变体，实现跨突触的活动依赖性、多色荧光重建 (X-RASP)。我们的系统首次允许根据突触（而不是整个神经元）的活动，以多种颜色对同一动物进行回顾性标记。由于单个突触通常充当大脑中的计算单元，因此我们的方法将促进使用现有技术无法实现的实验设计。此外，我们的策略很容易适应任何遗传系统中的电路图谱。 绘制电路内突触活动的能力将有助于阐明行为的神经基础。在这里，麦克弗森等人。报告用多色荧光标签专门标记果蝇中活跃突触的新策略。
Determining the pattern of activity of individual connections within a neural circuit could provide insights into the computational processes that underlie brain function. Here, we develop new strategies to label active synapses by trans-synaptic fluorescence complementation in Drosophila. First, we demonstrate that a synaptobrevin-GRASP chimera functions as a powerful activity-dependent marker for synapses in vivo. Next, we create cyan and yellow variants, achieving activity-dependent, multi-colour fluorescence reconstitution across synapses (X-RASP). Our system allows for the first time retrospective labelling of synapses (rather than whole neurons) based on their activity, in multiple colours, in the same animal. As individual synapses often act as computational units in the brain, our method will promote the design of experiments that are not possible using existing techniques. Moreover, our strategies are easily adaptable to circuit mapping in any genetic system. The ability to map the activity of synapses within a circuit will help in elucidating the neural basis of behaviour. Here, Macpherson et al. report new strategies to specifically label active synapses in Drosophila with multi-colour fluorescence tags.