Monitoring rapid guanosine signaling during ischemia

监测缺血期间的快速鸟苷信号传导

• 批准号: 10331885
• 负责人: Ashley E Ross
• 金额: $ 37.6万
• 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/10331885
• 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.

项目摘要 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， 现有的技术，但知识的动力学，调节机制，和功能的地方鸟苷 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， 然而，到目前为止，鸟苷在脑中的作用机制和动力学仍然是未知的。 悬而未决此外，了解鸟苷信号作为缺血功能的变化程度， 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 通过开发一种新的方法， 微流体平台，以在脑切片的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）记录鸟苷与毫秒到秒的时间分辨率， 对鸟苷调节的机制提供了重要的见解。测量局部鸟苷动力学在 具有显著改善的时空分辨率的损伤部位将提供大脑的关键信息， 立即响应当地的破坏。这一建议符合我们开发分析工具以检测 并了解损伤期间大脑中动态神经化学调节的炎症。这样做的理由 建议是，这些工具将提供动力学知识，机制，和功能的快速鸟苷 第一次在缺血期间大脑中发出信号，这可能进一步告知鸟苷的发展， 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） 开发用于向脑递送时空控制和持续局灶性缺血的微流体平台 （2）探讨海马区鸟苷快速释放和清除的机制， 缺血的严重程度和位置，以及（3）表征快速鸟苷信号对局部腺苷的影响 局部缺血时的变化。我们将通过使用新的微流体技术， 用于将缺血时间控制地传递到脑切片的平台， 鸟苷信号转导与FSCV的关系。这项工作意义重大，因为这些研究将使 对大脑在不同缺血持续时间内对缺血的即时反应的非凡机械见解 和严重程度，这将直接影响未来的脑损伤治疗策略。这些工具可以翻译成 任何生物系统来研究局部组织反应。预期的结果是一个新的平台， 第一次在大脑中的内源性鸟苷信号传导和鸟苷的深入了解 调节和神经调节。这项工作将对鸟苷的研究产生积极的影响 并将大大推进鸟苷在大脑即时损伤反应中的作用的知识。