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.

大脑功能由神经元的复杂协调网络执行，其操作模式为 对神经调节剂的星座高度敏感。更具体地，神经调节剂，例如多巴胺， 去甲肾上腺素，5-羟色胺和乙酰胆碱对全球脑过程的巨大控制（例如 唤醒，注意力，情感或认知感知。神经调节剂信号的改变已连接到 帕金森氏病，精神分裂症，抑郁和成瘾等神经和精神疾病。 同样，阿片类神经肽在认知和行为的调节中起重要作用。而 产生神经调节信号的解剖结构是众所周知的，对空间知之甚少 这些信号在神经支配的大脑区域中的时间演变。这是因为当前的测量 技术，例如微透析或环状伏安法，缺乏空间或时间分辨率（通常是 分子特异性）以解决各自的信号。这种技术挑战一直是一个长期的障碍 为了理解神经调节如何改变神经回路功能以影响行为。到 应对这一挑战，该项目将开发，验证和传播新的遗传编码 单胺神经调节剂和阿片类药物的大规模光学测量的荧光蛋白 通过将一个多学科的研究人员组合在一起，神经肽信号传导 独特而互补的专业知识。这些传感器蛋白有可能改变神经科学 以类似于钙，谷氨酸和电压的遗传编码指标，现在 广泛使用。结合钙和电压成像，神经调节器传感器将揭示这些方式 系统影响细胞和电路功能。特别是，提出的传感器将使最小的侵入性， 突触时神经调节剂和神经肽信号的高分辨率，长期和直接测量， 细胞和系统水平。长期以来，神经调节信号传导的传感器一直难以捉摸。我们的 Team最近开发了第一代的5-羟色胺（5-HT）的遗传编码指标， 去甲肾上腺素（NE）和多巴胺（DA）可以报告纳米摩尔浓度的变化，高 空间和时间分辨率。在这项工作的基础上，提出了以下特定目标：1）优化 并使用计算建模为单胺多样化的遗传编码传感器，定向 进化和高通量筛选； 2）开发和优化遗传编码的传感器以进行阿片类药物 使用新型蛋白支架的神经肽； 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。这项工作将共同提供 神经科学社区，具有广泛特征的多色指标，用于探测 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 健康与疾病。