Monitoring rapid guanosine signaling during ischemia

监测缺血期间的快速鸟苷信号传导

• 批准号: 10182458
• 负责人: Ashley E Ross
• 金额: $ 37.22万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10182458
• 关键词: Adenosine; Brain; Brain Injuries; Coupled; Development; Devices; Fostering; Frequencies; Future; Glucose; Goals; Guanosine; Health; Hippocampus (Brain); Hypoxia; Infarction; Inflammation; Injury; Ischemia; Knowledge; Length; Location; Measures; Methods; Microelectrodes; Microfluidic Microchips; Microfluidics; Mission; Monitor; Nervous System Trauma; Neuronal Injury; Outcome; Oxygen; Parahippocampal Gyrus; Periodicity; Play; Public Health; Purine Nucleosides; Purines; Regulation; Research; Resolution; Role; Scanning; Severities; Signal Transduction; Site; Slice; Techniques; Technology; Testing; Therapeutic; Time; Tissues; United States National Institutes of Health; Work; analytical tool; biological systems; carbon fiber; dentate gyrus; deprivation; design; improved; in vivo; innovation; insight; ischemic injury; millisecond; neurochemistry; neuroprotection; neuroregulation; novel; programs; receptor; response; spatiotemporal; targeted treatment; temporal measurement; therapeutic target; tool

PROJECT SUMMARY Measuring dynamic guanosine signaling at the site of focal ischemia over time remains challenging to probe with existing technology yet knowledge of the dynamics, regulatory mechanisms, and function of local guanosine fluctuations during ischemia would positively impact our understanding of the brain’s immediate local neuroprotective response. Guanosine is a nucleoside purine which has been postulated to play a potent restorative role after ischemic injury; however, to date, the mechanism and dynamics of guanosine action in the brain remains unresolved. Additionally, knowledge of the extent to which guanosine signaling changes as a function of ischemia duration and severity would provide critical insight into guanosine’s role as a neuroprotector. We propose to solve a significant gap in the understanding of guanosine signaling dynamics during focal ischemia by developing a microfluidic platform to initiate sustained local oxygen-glucose deprivation in a sub-region of a brain slice and using fast-scan cyclic voltammetry (FSCV) recording of guanosine with millisecond-to-second temporal resolution to provide critical insight into the mechanisms of guanosine regulation. Measuring local guanosine dynamics at the site of injury with significantly improved spatiotemporal resolution will provide critical information of the brain’s immediate local damage response. This proposal fits within our long-term goal to develop analytical tools to detect and understand dynamic neurochemical-regulated inflammation in the brain during injury. The rationale for this proposal is that these tools will provide knowledge of the dynamics, mechanism, and function of rapid guanosine signaling in the brain during ischemia for the first time which could further inform the development of guanosine- targeted therapies for neurological injury. The proposal will be completed by the following three specific aims: (1) Develop microfluidic platforms for delivery of spatiotemporally controlled and sustained focal ischemia to brain slices, (2) Characterize the mechanism of rapid guanosine release and clearance in the hippocampus as a function of ischemia severity and location, and (3) Characterize the impact of rapid guanosine signaling on local adenosine changes during focal ischemia. We will pursue these aims with an innovative approach by using novel microfluidic platforms for time-controlled delivery of ischemia to brain slices coupled to rapid electrochemical recording of guanosine signaling with FSCV for the first time. This work is significant because these studies will enable extraordinary mechanistic insight into the brain’s immediate response to ischemia over varying ischemia durations and severities which will directly impact future therapeutic strategies for brain injury. The tools are translatable to any biological system to study local tissue responses. The expected outcome is a new platform to investigate rapid endogenous guanosine signaling in the brain for the first time and an in-depth understanding of guanosine regulation and neuromodulation during ischemia. This work will have a positive impact on how guanosine is studied and will significantly advance knowledge of guanosine’s role in the brains immediate damage response.

项目概要 随着时间的推移，测量局灶性缺血部位的动态鸟苷信号传导仍然具有挑战性 现有技术以及对本地鸟苷的动力学、调节机制和功能的了解 缺血期间的波动将积极影响我们对大脑直接局部的理解 神经保护反应。鸟苷是一种核苷嘌呤，被认为具有有效的恢复作用 缺血性损伤后的作用；然而，迄今为止，鸟苷在大脑中作用的机制和动力学仍然存在 未解决。此外，了解鸟苷信号传导随缺血而变化的程度 持续时间和严重程度将为鸟苷作为神经保护剂的作用提供重要的见解。我们建议解决 通过开发一种方法，对局灶性缺血期间鸟苷信号动力学的理解存在显着差距 微流体平台在脑切片的一个子区域中启动持续的局部氧葡萄糖剥夺并使用 快速扫描循环伏安法（FSCV）以毫秒至秒的时间分辨率记录鸟苷 提供对鸟苷调节机制的重要见解。测量局部鸟苷动态 时空分辨率显着提高的损伤部位将提供大脑的关键信息 立即进行局部损害响应。该提案符合我们开发分析工具来检测的长期目标 并了解受伤期间大脑中动态神经化学调节的炎症。这样做的理由 建议这些工具将提供快速鸟苷的动力学、机制和功能的知识 首次在缺血期间大脑中发出信号，这可以进一步为鸟苷的发育提供信息 神经损伤的靶向治疗。该提案将通过以下三个具体目标来完成：（1） 开发微流体平台，用于向大脑输送时空控制和持续的局灶性缺血 切片，(2) 将海马中鸟苷快速释放和清除的机制表征为函数 缺血严重程度和位置，以及 (3) 表征快速鸟苷信号传导对局部腺苷的影响 局灶性缺血时的变化。我们将通过使用新型微流体以创新方法实现这些目标 用于时间控制地将缺血传递到脑切片并结合快速电化学记录的平台 首次使用 FSCV 进行鸟苷信号传导。这项工作意义重大，因为这些研究将使 对不同缺血持续时间内大脑对缺血的即时反应的非凡机制洞察 以及严重程度将直接影响未来脑损伤的治疗策略。这些工具可翻译为 研究局部组织反应的任何生物系统。预期结果是建立一个快速调查的新平台 首次研究大脑内源性鸟苷信号传导并深入了解鸟苷 缺血期间的调节和神经调节。这项工作将对鸟苷的研究产生积极影响 并将显着增进人们对鸟苷在大脑即时损伤反应中作用的认识。