Genetically encoded indicators for large-scale sensing of neuromodulatory signaling in behaving animals

用于大规模感知行为动物神经调节信号的基因编码指标

• 批准号: 9533713
• 负责人: Axel Nimmerjahn
• 金额: $ 95.31万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9533713
• 关键词: Acetylcholine; Acute; Address; Affect; Affinity; Amino Acids; Anatomy; Animal Behavior; Animals; Arousal; Arrestins; Attention; Behavior; Behavioral; Biological Assay; Body Regions; Brain; Brain region; Calcium; Catecholamines; Cell Nucleus; Cell Survival; Cells; Cognition; Cognitive; Color; Communication; Communities; Computer Simulation; Corpus striatum structure; Cyclic AMP; Dimensions; Directed Molecular Evolution; Disease; Dopamine; Dorsal; Electric Stimulation; Electrophysiology (science); Emotions; Evolution; Flow Cytometry; G-Protein-Coupled Receptors; GTP-Binding Proteins; Generations; Glutamates; Health; Human; Image; In Vitro; Kinetics; Lead; Ligands; Link; Measurement; Mental Depression; Mental disorders; Microdialysis; Microscopy; Molecular; Mus; Nervous system structure; Neurodegenerative Disorders; Neuroglia; Neuromodulator; Neurons; Neuropeptides; Neurosciences; Noise; Norepinephrine; Opioid; Optics; Output; Parkinson Disease; Pathway interactions; Pattern; Peptide Signal Sequences; Perception; Periodicity; Pharmaceutical Preparations; Play; Population; Process; Property; Proteins; Reporting; Research Personnel; Resolution; Role; Scaffolding Protein; Schizophrenia; Sensitivity and Specificity; Serotonin; Signal Transduction; Site; Site-Directed Mutagenesis; Slice; Specificity; Spinal; Spinal cord injury; Substantia nigra structure; Surface; Synapses; Synaptic Transmission; System; Techniques; Testing; Time; Validation; Viral; Whole-Cell Recordings; Work; addiction; arrestin 2; base; behavior influence; behavior test; beta-arrestin; calcium indicator; dorsal horn; drug of abuse; extracellular; gamma-Aminobutyric Acid; high throughput screening; in vivo; insight; locus ceruleus structure; minimally invasive; monoamine; mouse model; multidisciplinary; nano; nervous system disorder; neural circuit; neuronal patterning; neuroregulation; novel; operation; recruit; sensor; spatiotemporal; success; temporal measurement; trafficking; two-photon; virtual; voltage

Brain functions are executed by intricately coordinated networks of neurons, whose modes of operation are highly sensitive to a constellation of neuromodulators. More specifically, neuromodulators such as dopamine, norepinephrine, serotonin, and acetylcholine exert dramatic control over global brain processes such as arousal, attention, emotion, or cognitive perception. Altered neuromodulator signaling has been linked to neurological and psychiatric disorders such as Parkinson's disease, schizophrenia, depression and addiction. Similarly, opioid neuropeptides play important roles in the modulation of cognition and behavior. While the anatomical structures that produce neuromodulatory signals are well known, little is known about the spatial and temporal evolution of these signals in the innervated brain regions. This is because current measurement techniques, such as microdialysis or cyclic voltammetry, lack the spatial or temporal resolution (and often the molecular specificity) to resolve respective signals. This technical challenge has been a long-standing barrier to our understanding of how neuromodulation alters neural circuit function in order to influence behavior. To address this challenge, this project will develop, validate, and disseminate novel genetically encoded fluorescent proteins for large-scale optical measurement of monoamine neuromodulator and opioid neuropeptide signaling in behaving animals, by bringing together a multi-disciplinary team of investigators with unique and complementary expertise. These sensor proteins have the potential to revolutionize neuroscience in a way similar to genetically encoded indicators for calcium, glutamate, and voltage, which are now in widespread use. Combined with calcium and voltage imaging, neuromodulator sensors will reveal how these systems impinge on cellular and circuit function. In particular, proposed sensors will enable minimally invasive, high-resolution, long-term, and direct measurement of neuromodulator and neuropeptide signaling at synaptic, cellular, and system levels. Sensors for neuromodulatory signaling have remained elusive for a long time. Our team recently developed a first generation of genetically encoded indicators for serotonin (5-HT), norepinephrine (NE), and dopamine (DA) that can report nano- to micromolar concentration changes with high spatial and temporal resolution. Building on this work, the following specific aims are proposed: 1) Optimize and diversify genetically encoded sensors for the monoamines using computational modeling, directed evolution and high-throughput screening; 2) Develop and optimize genetically encoded sensors for opiate neuropeptides using novel protein scaffolds; and 3) Systematically validate the novel neuromodulator and neuropeptide sensors in acute brain slices and behaving animals. Together, this work will provide the neuroscience community with a wide range of well-characterized multi-color indicators for probing the functional role of neuromodulators and neuropeptides in regulating neural circuit function and behavior in both health and disease.

大脑功能是由复杂协调的神经元网络执行的，其操作模式是 对一系列神经调节剂高度敏感。更具体地说，神经调节剂如多巴胺、 去甲肾上腺素、血清素和乙酰胆碱对大脑的整体过程发挥着巨大的控制作用，例如 唤醒、注意力、情绪或认知知觉。神经调节信号的改变与 神经和精神疾病，如帕金森病、精神分裂症、抑郁症和成瘾症。 同样，阿片类神经肽在认知和行为的调节中发挥着重要作用。虽然 产生神经调节信号的解剖结构是众所周知的，但人们对空间神经调节信号知之甚少。 以及这些信号在受神经支配的大脑区域中的时间演变。这是因为电流测量 微透析或循环伏安法等技术缺乏空间或时间分辨率（并且通常 分子特异性）来解析各自的信号。这一技术挑战一直是一个长期存在的障碍 帮助我们理解神经调节如何改变神经回路功能以影响行为。到 为了应对这一挑战，该项目将开发、验证和传播新型基因编码 用于大规模光学测量单胺神经调节剂和阿片类药物的荧光蛋白 通过将多学科研究人员团队聚集在一起，研究行为动物中的神经肽信号传导 独特且互补的专业知识。这些传感器蛋白有可能彻底改变神经科学 以类似于钙、谷氨酸和电压的基因编码指标的方式，这些指标现在已在 广泛使用。与钙和电压成像相结合，神经调节器传感器将揭示这些 系统会影响细胞和电路功能。特别是，所提出的传感器将实现微创、 高分辨率、长期、直接测量突触处的神经调节剂和神经肽信号传导， 蜂窝和系统级别。神经调节信号传感器长期以来一直难以捉摸。我们的 团队最近开发了第一代血清素（5-HT）基因编码指标， 去甲肾上腺素 (NE) 和多巴胺 (DA) 可以报告纳摩尔至微摩尔的浓度变化，并且具有高 空间和时间分辨率。在此工作的基础上，提出以下具体目标： 1）优化 并使用计算模型使单胺基因编码传感器多样化，定向 进化和高通量筛选； 2) 开发和优化阿片类药物基因编码传感器 使用新型蛋白质支架的神经肽； 3）系统地验证新型神经调节剂和 急性脑切片和行为动物中的神经肽传感器。总之，这项工作将提供 神经科学界拥有广泛的特征明确的多色指标，用于探索 神经调节剂和神经肽在调节神经回路功能和行为中的功能作用 健康和疾病。