A Genetically Encoded Fluorescent Sensor for Rapid and Specific In Vivo Detection of Norepinephrine
A Genetically Encoded Fluorescent Sensor for Rapid and Specific In Vivo Detection of Norepinephrine
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DOI：
10.1016/j.neuron.2019.02.037
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发表时间：
2019-05-22
期刊：
影响因子：
16.2
通讯作者：
Li, Yulong
中科院分区：
文献类型：
--
作者：
Feng, Jiesi;Zhang, Changmei;Li, Yulong
去甲肾上腺素（NE）是一种重要的生物单胺类神经递质，参与多种生理过程。然而，其精确的动力学和调节仍然很差的特点，部分原因是现有的技术在体内测量NE的限制。在这里，我们开发了一系列基于GPCR激活的NE（GRAB（NE））传感器，对NE具有230%的峰值Delta F/F-0响应、良好的光稳定性、纳摩尔至微摩尔灵敏度、亚秒级动力学和高特异性。病毒或转基因介导的GRAB（NE）传感器的表达能够检测小鼠脑切片蓝斑（LC）中的电刺激诱发的NE释放，活斑马鱼中脑中的隐现诱发的NE释放，以及自由移动小鼠LC和下丘脑中的光遗传学和行为触发的NE释放。因此，GRAB（NE）传感器是用于快速和特异性监测生理和病理过程中体内NE传输的强大工具。
Norepinephrine (NE) is a key biogenic monoamine neurotransmitter involved in a wide range of physiological processes. However, its precise dynamics and regulation remain poorly characterized, in part due to limitations of available techniques for measuring NE in vivo. Here, we developed a family of GPCR activation-based NE (GRAB(NE)) sensors with a 230% peak Delta F/F-0 response to NE, good photostability, nanomolar-to-micromolar sensitivities, sub-second kinetics, and high specificity. Viral- or transgenic-mediated expression of GRAB(NE) sensors was able to detect electrical-stimulation-evoked NE release in the locus coeruleus (LC) of mouse brain slices, looming-evoked NE release in the midbrain of live zebrafish, as well as optogenetically and behaviorally triggered NE release in the LC and hypothalamus of freely moving mice. Thus, GRAB(NE) sensors are robust tools for rapid and specific monitoring of in vivo NE transmission in both physiological and pathological processes.