Optogenetic reporters: Fluorescent protein-based genetically encoded indicators of signaling and metabolism in the brain.
Optogenetic reporters: Fluorescent protein-based genetically encoded indicators of signaling and metabolism in the brain.
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DOI：
10.1016/b978-0-444-59426-6.00012-4
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发表时间：
2012
影响因子：
--
通讯作者：
Yellen, Gary
中科院分区：
文献类型：
--
作者：
Tantama, Mathew;Hung, Yin Pun;Yellen, Gary
关键词：
荧光蛋白质技术已经发展到包括基因编码的生物传感器，可以监测离子水平、代谢物、酶活性以及蛋白质构象，甚至膜电压。它们非常适合于活细胞显微镜和定量分析，并且可以用于多种成像模式，包括单光子或双光子荧光强度或寿命显微镜。虽然还不是很完整，但现在已经有大量的遗传编码的报告，可以用来监测神经元和神经胶质生物学的许多方面，这些生物传感器可以用来可视化突触传递和活动依赖的信号在体外和体内。在这篇综述中，我们介绍了工程生物传感器的设计策略，包括使用循环排列的荧光蛋白设计传感器和利用荧光蛋白之间的荧光共振能量转移(FRET)设计传感器。我们还提供了感知小离子(例如，pH、氯、锌)、代谢物(例如，谷氨酸、葡萄糖、三磷酸腺苷、cAMP、脂类代谢物)、信号通路(例如，G蛋白偶联受体、Rho GTP酶)、酶活性(例如，蛋白激酶A、半胱氨酸天冬氨酸氨基转移酶)和活性物种的指示剂的例子。我们的重点是这些基因编码的指示物被应用于与大脑相关的研究并与活细胞荧光显微镜一起使用的例子。
Fluorescent protein technology has evolved to include genetically-encoded biosensors that can monitor levels of ions, metabolites, and enzyme activities as well as protein conformation and even membrane voltage. They are well suited to live-cell microscopy and quantitative analysis, and they can be used in multiple imaging modes, including one or two-photon fluorescence intensity or lifetime microscopy. Although not nearly complete, there now exists a substantial set of genetically-encoded reporters that can be used to monitor many aspects of neuronal and glial biology, and these biosensors can be used to visualize synaptic transmission and activity-dependent signaling in vitro and in vivo. In this review we present an overview of design strategies for engineering biosensors, including sensor designs using circularly-permuted fluorescent proteins and using fluorescence resonance energy transfer (FRET) between fluorescent proteins. We also provide examples of indicators that sense small ions (e.g., pH, chloride, zinc), metabolites (e.g., glutamate, glucose, ATP, cAMP, lipid metabolites), signaling pathways (e.g., G protein coupled receptors, Rho GTPases), enzyme activities (e.g., protein kinase A, caspases), and reactive species. We focus on examples where these genetically-encoded indicators have been applied to brain-related studies and used with live-cell fluorescence microscopy.