Multiplexed Sensing and Control of Neuromodulators and Peptides in the Awake Brain

清醒大脑中神经调节剂和肽的多重传感和控制

• 批准号: 10731789
• 负责人: Mark L Andermann
• 金额: $ 25.94万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10731789
• 关键词: 3-Dimensional; Acetylcholine; Acute; Adenosine; Arousal; Axon; Bathing; Behavioral; Boston; Brain; Brain region; Calibration; Cannulas; Cells; Cerebrospinal Fluid; Chemicals; Child; Chronic; Clinical; Closure by clamp; Cognitive; Collaborations; Complement; Corticotropin-Releasing Hormone; Cultured Cells; Disease; Dopamine; Dose; Drug Delivery Systems; Exhibits; Fiber; Fiber Optics; Fluorescence; Green Fluorescent Proteins; Harvest; Head; Histamine; Hour; Hydrogels; Hypothalamic structure; Image; Implant; Individual; Liquid substance; Locomotion; Measurement; Measures; Medial; Melatonin; Mental disorders; Methods; Microdialysis; Microfluidics; Monitor; Mus; Nature; Neuromodulator; Neurons; Neurosciences; Neurosciences Research; Norepinephrine; Oxytocin; Pattern; Peptides; Pharmaceutical Preparations; Photometry; Photons; Preoptic Areas; Refractive Indices; Retinal blind spot; Robotics; Serotonin; Signal Transduction; Somatostatin; Technology; Testing; Time; Vasoactive Intestinal Peptide; Vasopressins; awake; behavior test; brain cell; brain tissue; cellular imaging; cost effective; experimental study; fluorescence imaging; high dimensionality; improved; in vivo; intraperitoneal; lateral ventricle; lens; nervous system disorder; neural patterning; neuropsychiatry; neuroregulation; novel; optical fiber; optical sensor; optogenetics; preference; quantitative imaging; rational design; response; sensor; side effect; social; tool; two-photon; virtual reality

Summary Imbalanced levels of neuromodulators and other chemical signals contribute to a host of neurological disorders. Yet, previous studies describing these effects often examine only one molecule at a time, and typically provide a static description of signal levels in the brain or in the cerebrospinal fluid (CSF) that bathes all neurons. In reality, dozens of signals exhibit dynamic changes across states such as quiet waking and social or non-social arousal, which are altered in disease. The tracking and manipulation of patterns of neural activity has been critical to recent neuroscience progress. We lack analogous tools for estimation and control of dynamic patterns of neuromodulatory signals, which could revolutionize the study of brain states and effectively restore healthy states across neurologic and psychiatric disorders. Moreover, we do not understand how any given neuropsychiatric drug dynamically influences the levels of endogenous neuromodulators and peptides in the CSF or brain, thus impeding the rational design of optimal drug delivery strategies to maximize efficacy and minimize side effects. These blind spots are due to technical limitations: while cellular imaging and optogenetics have enabled ever-increasing precision in tracking and manipulation of brain cells, we lack the ability to accurately (i) record or (ii) control multiple neuromodulatory signals simultaneously in real time. We are overcoming the first challenge by developing novel methods for multiplexed, quantitative imaging of a panel of green fluorescent protein-based optical sensors of disease-relevant neuromodulatory signals (Aim 1): vasopressin, oxytocin, somatostatin, dopamine, norepinephrine, serotonin, acetylcholine, histamine, melatonin, corticotropin-releasing factor, vasoactive intestinal peptide, and adenosine. Briefly, sets of cultured cells expressing individual sensors are combined in a 3D hydrogel sensor array applied to the front of a gradient refractive index (GRIN) lens, which is inserted into the CSF or brain tissue of an awake, head-fixed mouse via a chronic cannula. Estimates of signal concentration using 3D two-photon imaging of the sensor array are then calibrated via post-hoc robotic dipping of the same sensor array into varying concentrations of each neuromodulator ex vivo. Once we have established this approach to track neuromodulatory composition across hours or days and across behavioral states (Aim 1), we will use closed-loop delivery methods to control dynamic patterns of up to a dozen neuromodulatory signals in the brain in awake mice and evaluate which patterns drive behavioral preference or avoidance (Aim 2).These experiments benefit from the use of fluorescence lifetime and well as fluorescence intensity measurements, allowing quantitative assessment of fluid composition across extended periods of time (hours to days) with minimal effects of bleaching. Together, these tools offer a novel, holistic framework for the study and control of multiple neuromodulators in the brain. The sensitive, real-time, multiplexed readout of signals in small volumes complements microdialysis and enables closed-loop control with applications to most domains of basic and clinical neuroscience research.

概括 神经调节剂和其他化学信号水平不平衡会导致一系列神经系统疾病。 然而，之前描述这些效应的研究通常一次只检查一个分子，并且通常提供 对大脑或沐浴所有神经元的脑脊液 (CSF) 中信号水平的静态描述。在 现实中，数十种信号在不同状态下表现出动态变化，例如安静醒来、社交或非社交 觉醒，在疾病中会改变。神经活动模式的跟踪和操纵已经被 对最近的神经科学进展至关重要。我们缺乏类似的工具来估计和控制动态模式 神经调节信号，这可能会彻底改变大脑状态的研究并有效恢复健康 神经和精神疾病的状态。此外，我们不明白任何给定的 神经精神药物动态影响内源性神经调节剂和肽的水平 CSF 或大脑，从而阻碍了最佳药物输送策略的合理设计，以最大限度地提高疗效和 最大限度地减少副作用。这些盲点是由于技术限制造成的：而细胞成像和光遗传学 脑细胞的追踪和操纵精度不断提高，但我们缺乏能力 实时准确地（i）记录或（ii）同时控制多个神经调节信号。我们是 通过开发对一组细胞进行多重定量成像的新方法来克服第一个挑战 基于绿色荧光蛋白的疾病相关神经调节信号光学传感器（目标 1）： 加压素、催产素、生长抑素、多巴胺、去甲肾上腺素、血清素、乙酰胆碱、组胺、褪黑激素、 促肾上腺皮质激素释放因子、血管活性肠肽和腺苷。简而言之，培养细胞组 表示各个传感器组合在 3D 水凝胶传感器阵列中，应用于梯度的前面 折射率（GRIN）透镜，通过一个固定头部的清醒小鼠的脑脊液或脑组织 慢性插管。然后使用传感器阵列的 3D 双光子成像来估计信号浓度 通过事后机器人将同一传感器阵列浸入不同浓度的每种传感器中进行校准 离体神经调节剂。一旦我们建立了这种方法来追踪神经调节成分 小时或天以及跨行为状态（目标 1），我们将使用闭环交付方法来控制动态 清醒小鼠大脑中多达十几种神经调节信号的模式，并评估哪些模式驱动 行为偏好或回避（目标 2）。这些实验受益于荧光寿命和 以及荧光强度测量，可以定量评估整个液体成分 长时间（几小时到几天）漂白效果最小。这些工具共同提供了一种新颖的、 研究和控制大脑中多种神经调节剂的整体框架。敏感的人， 小体积信号的实时、多重读出补充了微透析并使得 闭环控制，适用于基础和临床神经科学研究的大多数领域。