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

基本信息

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
项目类别：
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

项目摘要

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）：该提案的目标是开发生物胺的基因编码指示剂工具箱，生物胺是最重要的神经调节剂家族，所有神经系统都受到神经调节的影响，神经调节通过改变神经回路的动力学来重新配置。目标神经元的内在放电特性并调节其突触可塑性，神经调节剂的动力学改变与许多人类神经和精神疾病有关，包括帕金森病、精神分裂症和成瘾。生物胺是所有动物大脑用来调节神经回路的发育、结构和功能的一组神经调节剂，尽管人们对生物胺投射的解剖学特征和功能意义有一定程度的了解，但其确切机制尚不清楚。这些分子对行为的控制尚不完全清楚，为了破译这些分子对大脑和行为产生影响的机制，我们必须对神经调节剂瞬变进行敏感和具体的测量，无论是广泛的还是广泛的。（体积调制）和局部（目标调制），具有必要的空间和时间分辨率，理想情况下在 intac 电路中，包括微透析和循环伏安法，是有用的，但不足以完成当前的任务。开发基于荧光蛋白的基因编码指示剂，结合现代显微镜，可以直接、特异性地测量不同类型的神经调节剂，并具有增强的空间和时间分辨率。最近，我们成功建立了开发技术平台。神经活动的基因编码指标，基于高度优化的传感器传感器平台和神经科学传感器表征和应用的丰富经验，开发出了几种高质量的光学探针，用于对活体动物中的大规模神经群体进行同步成像。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，特别是对于行为最普遍的神经调节剂多巴胺，我们的具体目标将从使用组合计算设计和筛选每种生物胺的传感器开始。然后，我们将开发突触靶向策略来显示树突和轴突中的传感器，以提高其突触成像的实用性。这些传感器在活体神经元和大鼠脑切片中的性能，并证明它们在活体动物中探测多巴胺瞬变动态的能力，这些分子的最先进的传感器将促进非侵入性、精确、直接和连续的测量。鉴于生物源的明确相关性，光学记录方面的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方面的动态图景。由于胺对神经系统疾病的影响，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。