Monitoring neurochemical signaling dynamics in the lymph node

监测淋巴结中的神经化学信号动态

• 批准号: 10460564
• 负责人: Ashley E Ross
• 金额: $ 38.79万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-14 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10460564
• 关键词: Anions; Autoimmunity; Brain; Buffers; Carbon; Catecholamines; Cell Communication; Cells; Chemistry; Chronic; Communication; Data; Dendritic Cells; Detection; Development; Disease; Electrodes; Fostering; Goals; Health; Immune; Immune response; Immune system; Immunity; Immunomodulators; Immunotherapy; Inflammation; Knowledge; Lead; Lymph Node Tissue; Malignant Neoplasms; Measurement; Measures; Melatonin; Mental Depression; Methods; Microelectrodes; Microfluidics; Mission; Monitor; Neurohormones; Neuroimmune; Neuroimmunomodulation; Neurons; Neurophysiology - biologic function; Neurotransmitters; Nitrogen; Norepinephrine; Organ; Outcome; Periodicity; Peripheral; Public Health; Regulation; Research; Resistance; Resolution; Scanning; Signal Transduction; Slice; Surface; TNF gene; Technology; Testing; Time; Tissues; United States National Institutes of Health; Work; analytical tool; base; biological systems; carbon fiber; gastrointestinal; gut-brain axis; immunoregulation; improved; innovation; lymph nodes; mesenteric lymph node; multiplex detection; nerve supply; neurochemistry; programs; real time monitoring; receptor; sensor; spatiotemporal; targeted treatment; temporal measurement; tool

PROJECT SUMMARY Neurochemical signaling within immune organs, like the lymph node, remains challenging to probe with existing technology yet knowing the mechanisms and function of this signaling would positively impact our understanding of immunity. Our long-term goal is to understand neurochemical regulated immunity during inflammation, autoimmunity, and even depression. To achieve this goal, new analytical tools are needed which can capture rapid neurochemical signaling in intact immune organs with high spatial resolution. The specific objective of this proposal is to develop and validate methods using fast-scan cyclic voltammetry (FSCV) at carbon-based microelectrodes to detect norepinephrine, ATP, and melatonin in slices of the mesenteric lymph node (mLN). All three neurochemicals are important for either triggering or suppressing immune responses within the gut-immune system; however, the dynamics and mechanisms by which they function are not understood. The rationale for this proposal is that the development of new tools to monitor rapid neurochemical signaling in an intact mLN will provide knowledge of neuroimmune communication dynamics in the gut which could lead to sophisticated neurochemical-targeted therapies for gastrointestinal inflammation and an improved understanding of the gut-brain axis. The proposal will be completed by the following three specific aims: (1) Develop innovative electrochemical methods to detect and validate neuronal norepinephrine release in live mLN slices, (2) Develop anion-exchange doped carbon-fiber microelectrodes for sensitive ATP detection in the mLN, and (3) Develop fouling-resistant sensors for multiplexed detection of melatonin with catecholamines in the mLN. We will pursue these aims with an innovative approach combining the power of fast-scan cyclic voltammetry’s high temporal resolution and spatial resolution with detection in live slices of the lymph node. This work is also innovative because new carbon electrodes and surface chemistries will be developed for targeted-analyte detection. This work is significant because the tools developed will help shift the paradigm that immunomodulation is slow and will impact our understanding of neuroimmune communication mechanisms and dynamics, specifically within the gut-immune system. Tools to detect rapid concentrations fluctuations in norepinephrine, ATP, and melatonin are also significant because they are not only involved in immunomodulation in the immune system, but are heavily involved in signaling throughout the body. The tools are translatable to any biological system. The expected outcome is a new toolbox for high temporal resolution detection of neurochemicals in the lymph node which will lead to an improved understanding of the mechanism and function of neurochemical signaling in spatially-resolved regions of mLN during conditions of health and inflammation. This work will have a positive impact on how neuroimmune communication is studied, and will advance current knowledge of neurotransmitter regulated immunity leading to advancements in targeted immunotherapies for autoimmunity, cancer, and disease.

项目概要 免疫器官（如淋巴结）内的神经化学信号传导仍然具有挑战性，无法用现有技术进行探测 技术但了解这种信号传导的机制和功能将对我们的理解产生积极影响 我们的长期目标是了解炎症期间神经化学调节的免疫， 为了实现这一目标，需要新的分析工具来快速捕获。 具有高空间分辨率的完整免疫器官中的神经化学信号传导。 是开发和验证在碳基微电极上使用快速扫描循环伏安法（FSCV）的方法 检测肠系膜淋巴结 (mLN) 切片中的去甲肾上腺素、ATP 和褪黑激素 所有三种神经化学物质。 对于触发或抑制肠道免疫系统内的免疫反应很重要； 它们发挥作用的动力和机制尚不清楚。 开发新工具来监测完整 mLN 中的快速神经化学信号传导将提供以下知识： 肠道中的神经免疫通讯动态可能导致复杂的神经化学靶向 该提案将促进胃肠道炎症的治疗以及加深对肠脑轴的了解。 通过以下三个具体目标来完成：（1）开发创新的电化学方法来检测和 验证活 mLN 切片中神经元去甲肾上腺素的释放，(2) 开发阴离子交换掺杂碳纤维 用于 mLN 中灵敏 ATP 检测的微电极，以及 (3) 开发用于多路复用的防污传感器 检测 mLN 中的褪黑激素和儿茶酚胺 我们将通过创新方法实现这些目标。 将快速扫描循环伏安法的高时间分辨率和空间分辨率的强大功能与检测相结合 这项工作也具有创新性，因为新的碳电极和表面。 这项工作将开发用于目标分析物检测的化学方法，因为所开发的工具非常重要。 将有助于改变免疫调节缓慢的范式并将影响我们对神经免疫的理解 通讯机制和动态，特别是在肠道免疫系统内的快速检测工具。 去甲肾上腺素、ATP 和褪黑激素的浓度波动也很显着，因为它们不仅 参与免疫系统的免疫调节，但也大量参与全身的信号传导。 这些工具可应用于任何生物系统，预期结果是一个用于高时间的新工具箱。 分辨率检测淋巴结中的神经化学物质，这将有助于更好地了解 健康条件下 mLN 空间分辨区域神经化学信号传导的机制和功能 这项工作将对神经免疫通讯的研究产生积极影响，并将产生积极影响。 当前对神经递质调节免疫的了解导致了靶向治疗的进步 针对自身免疫、癌症和疾病的免疫疗法。