Departments of Biochemistry and Molecular Medicine, Chemistry, Statistics, Molecular and Cellular Biology, and Physiology and Membrane Biology, the Center for Neuroscience, and Graduate Programs in Molecular, Cellular, and Integrative Physiology, Biochemistry, Molecular, Cellular and Developmental Biology and Neuroscience, University of California, Davis, Davis, CA 95616, USA.
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20174, USA.
Cell. 2020 Dec 23;183(7):1986-2002.e26. doi: 10.1016/j.cell.2020.11.040. Epub 2020 Dec 16.
Serotonin plays a central role in cognition and is the target of most pharmaceuticals for psychiatric disorders. Existing drugs have limited efficacy; creation of improved versions will require better understanding of serotonergic circuitry, which has been hampered by our inability to monitor serotonin release and transport with high spatial and temporal resolution. We developed and applied a binding-pocket redesign strategy, guided by machine learning, to create a high-performance, soluble, fluorescent serotonin sensor (iSeroSnFR), enabling optical detection of millisecond-scale serotonin transients. We demonstrate that iSeroSnFR can be used to detect serotonin release in freely behaving mice during fear conditioning, social interaction, and sleep/wake transitions. We also developed a robust assay of serotonin transporter function and modulation by drugs. We expect that both machine-learning-guided binding-pocket redesign and iSeroSnFR will have broad utility for the development of other sensors and in vitro and in vivo serotonin detection, respectively.
血清素在认知中起着核心作用，是大多数精神疾病药物的靶点。现有的药物疗效有限；创造更好的版本将需要更好地了解血清素能电路，这一直受到我们无法以高时空分辨率监测血清素释放和转运的阻碍。我们开发并应用了一种由机器学习指导的结合口袋重新设计策略，创建了一种高性能、可溶性、荧光血清素传感器（iSeroSnFR），能够光学检测毫秒级的血清素瞬变。我们证明，iSeroSnFR 可用于在恐惧条件反射、社交互动和睡眠/觉醒转变期间检测自由行为小鼠中的血清素释放。我们还开发了一种用于检测血清素转运体功能和药物调节的可靠方法。我们预计，机器学习指导的结合口袋重新设计和 iSeroSnFR 将分别在其他传感器的开发以及体外和体内血清素检测方面具有广泛的用途。
