Genetically encoded sensors for the biogenic amines: watching neuromodulation in action

生物胺的基因编码传感器：观察神经调节的作用

• 批准号: 8934236
• 负责人: Lin Tian
• 金额: $ 38.85万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8934236
• 关键词: Amines; Animal Disease Models; Animal Model; Animals; Axon; Behavior; Behavioral; Binding; Biogenic Amines; Brain; California; Cell membrane; Dendrites; Dendritic Spines; Detection; Development; Diffusion; Distant; Dopamine; Epinephrine; Event; Family; Farming environment; Fishes; Foundations; Glutamates; Goals; Head; Health; Histamine; Human; Image; In Vitro; Individual; Kinetics; Life; Longitudinal Studies; Mammalian Cell; Maps; Measurement; Mental disorders; Methods; Microdialysis; Microscopy; Midbrain structure; Molecular; Mus; Myosin ATPase; Nervous system structure; Neuromodulator; Neurons; Neurosciences; Neurotransmitters; Noise; Norepinephrine; One-Step dentin bonding system; Optics; Paint; Parkinson Disease; Pattern; Performance; Periplasmic Binding Proteins; Population; Presynaptic Terminals; Property; Protein Engineering; Proteins; Rattus; Reporting; Resolution; Rodent; Schizophrenia; Sensitivity and Specificity; Serotonin; Signal Transduction; Slice; Solid; Solutions; Specificity; Structure; Surface; Synapses; Synaptic plasticity; System; Technology; Testing; Therapeutic; Time; Universities; Validation; Vertebral column; addiction; base; brain behavior; calcium indicator; cell type; cytotoxicity; design; directed evolution; experience; fly; high throughput screening; human stem cells; improved; in vivo; nervous system disorder; neural circuit; neuronal cell body; neuroregulation; neurotransmission; optical sensor; relating to nervous system; response; scaffold; screening; sensor; small molecule; success; temporal measurement; tool; treatment strategy; two-photon

 DESCRIPTION (provided by applicant): The goal of this proposal is to develop a toolbox of genetically encoded indicators for biogenic amines, the most important family of neuromodulators. All nervous systems are subject to neuromodulation, which reconfigure the dynamics of neural circuitry by transforming the intrinsic firing properties of targeted neurons and regulating their synaptic plasticity. The altered dynamics of the neuromodulators have been implicated in a number of human neurological and psychiatric diseases, including Parkinson's, schizophrenia and addiction. Biogenic amines are a group of neuromodulators used by all animal brains to regulate the development, structure and function of neural circuits. Although the anatomical characterization and functional significance of biogenic amine projections are understood to a moderate degree, the precise mechanisms by which these molecules exert control over behavior are not fully understood. To decipher the mechanisms by which these molecules exert their influence on the brain and behavior, we must perform sensitive and specific measurements of neuromodulator transients, both broadly (volume modulation) and locally (targeted modulation), with the requisite spatial and temporal resolution, ideally in intac circuits. Existing methods, encompassing microdialysis and cyclic voltammetry, are useful, but not adequate for this task at hand. One potential solution would be to develop genetically encoded indicators based on fluorescent proteins combined with modern microscopy allowing direct and specific measurement of diverse types of neuromodulators with enhanced spatial and temporal resolutions. Recently we have successfully established technology platform for the development of genetically encoded indicators of neural activity, which have led to several high-quality optical probes for simultaneous imaging of large-scale neuronal populations in living animals. Building upon highly optimized platform for sensor sensors and extensive experience in sensor characterization and application in neuroscience, we propose to develop a high-quality toolkit of optical sensors for the biogenic amine neuromodulators, especially for dopamine, the most behavioral pervasive neuromodulator. Our specific aims will start by designing and screening sensors for each of the biogenic amines using combined computational redesign and direct revolution. We will then develop synaptic targeting strategies to display the sensors in dendrites and axons to improve their utility for synaptic imaging. We will finally characterize the performance of these sensors in living neurons and in rat brain slices and demonstrate their capabilities of probing dynamics of dopamine transients in living animals. State-of-the-art sensors for these molecules will facilitate the non-invasive, precise, direct and continual measurement of released neuromodulators at both the synaptic and circuit levels in live model organisms. Such technology advance in optical recordings will facilitate neural circuitry mapping and paint a dynamic picture of neuromodulation systems in regulating neural circuitry and behavior. Given the clear relevance of the biogenic amines to the neurological diseases, these sensors are especially beneficial for long-term studies of human stem cell and animal disease models (specially the Parkinson's disease) and evaluating the effects of candidate therapeutics.

 描述（由申请人提供）： 这项提案的目标是开发一个工具箱的遗传编码指标生物胺，最重要的家庭的神经调质。所有的神经系统都受到神经调节的影响，神经调节通过改变目标神经元的固有放电特性并调节其突触可塑性来重新配置神经回路的动力学。神经调质的动力学改变与许多人类神经和精神疾病有关，包括帕金森氏症、精神分裂症和成瘾。生物胺是一组神经调质，被所有动物大脑用于调节神经回路的发育、结构和功能。虽然生物胺投射的解剖学特征和功能意义被理解到中等程度，但这些分子对行为施加控制的精确机制尚未完全理解。为了破译这些分子对大脑和行为产生影响的机制，我们必须对神经调质瞬变进行灵敏和特异性的测量，包括广泛的（体积调制）和局部的（靶向调制），具有必要的空间和时间分辨率，最好是在intac电路中。现有的方法，包括微透析和循环伏安法，是有用的，但不足以完成这项任务。一个潜在的解决方案是开发基于荧光蛋白的遗传编码指示剂，结合现代显微镜技术，以增强的空间和时间分辨率直接和特异性地测量各种类型的神经调质。最近，我们已经成功地建立了用于开发神经活动的遗传编码指标的技术平台，这导致了几种高质量的光学探针，用于活体动物中大规模神经元群体的同时成像。基于高度优化的传感器传感器平台和在传感器表征和神经科学应用方面的丰富经验，我们建议开发一种高质量的生物胺神经调节剂光学传感器工具包，特别是多巴胺，最普遍的行为神经调节剂。我们的具体目标将开始设计和筛选传感器的每一个生物胺使用相结合的计算重新设计和直接革命。然后，我们将开发突触靶向策略，以显示树突和轴突中的传感器，以提高它们在突触成像中的实用性。我们将最终描述 这些传感器在活神经元和大鼠脑切片中的性能，并证明了它们在活体动物中探测多巴胺瞬变动力学的能力。这些分子的最先进的传感器将有助于在活的模型生物体中的突触和回路水平上对释放的神经调质进行非侵入性、精确、直接和连续的测量。光学记录的这种技术进步将促进神经回路映射，并描绘神经调节系统在调节神经回路和行为中的动态图像。鉴于生物胺与神经系统疾病的明显相关性，这些传感器特别有利于人类干细胞和动物疾病模型（特别是帕金森病）的长期研究以及评估候选疗法的效果。