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（应用程序提供）：该提案的目的是开发一个针对生物胺的遗传编码指标的工具箱，这是最重要的神经调节剂家族。所有神经系统均受到神经调节的影响，该神经调节通过转换靶向神经元的内在发射特性并控制其合成可塑性来重新配置神经循环的动力学。神经调节剂的动力学改变了，包括帕金森氏症，精神分裂症和成瘾在内的许多人类神经和精神病疾病。生物胺是所有动物大脑使用的一组神经调节剂，用于调节神经环节的发育，结构和功能。尽管将生物胺项目的解剖表征和功能意义理解为现代程度，但这些分子对行为的控制的确切机制尚未完全理解。为了破译这些分子对大脑和行为的影响的机制，我们必须对神经调节剂瞬变进行敏感和特定的测量，无论是广泛的（体积调制）和局部（靶向调制），并具有必要的空间和临时分辨率，在INTAC电路中是理想的。现有的方法包括微透析和循环伏安法是有用的，但对于手头的任务不足。一种潜在的解决方案是基于荧光蛋白与现代显微镜结合的一般编码指标，从而可以直接和特定地测量使用增强的空间和临时分辨率的潜水员类型的神经调节剂。最近，我们成功地建立了技术平台，用于开发神经活性的遗传编码指标，这导致了几个高质量的光学问题，用于简单地对活动物中大规模神经元种群进行简单的成像。在高度优化的传感器传感器平台上，以及在神经科学中的传感器表征和应用方面的丰富经验，我们建议为生物胺神经调节剂开发高质量的光学传感器工具包，尤其是对多巴胺，尤其是行为上最普遍的神经调节剂。我们的具体目的将从使用计算重新设计和直接革命为每个生物胺设计和筛选传感器开始。然后，我们将开发突触靶向策略，以在树突和轴突中显示传感器，以改善其突触成像的效用。我们最终将表征 这些传感器在活性神经元和大鼠脑切片中的性能，并证明了它们在活动物中探测多巴胺瞬变动力学的能力。这些分子的最先进传感器将促进在活模型生物体中突触和电路水平上释放的神经调节剂的非侵入性，精确，直接和连续测量。光学记录中的这种技术进步将有助于神经元电路映射，并在调节神经元电路和行为中描绘神经调节系统的动态图片。鉴于生物胺对神经系统疾病的明显缓解，这些传感器对人类干细胞和动物病模型（特别是帕金森氏病）的长期研究特别有益，并评估了候选治疗的影响。