Multiplexed neurochemical methods to understand adenosine neuromodulation

多重神经化学方法了解腺苷神经调节

• 批准号: 10538604
• 负责人: B. JILL VENTON
• 金额: $ 60.17万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10538604
• 关键词: Acetylcholine; Adenosine; Biological; Brain; Calcium; Calcium Signaling; Cells; Color; Detection; Development; Diffusion; Disease; Disease model; Dopamine; Electrodes; Event; Fiber; Future; Genetic; Glutamates; Goals; Ischemia; Knowledge; Measurement; Measures; Mediating; Methods; Microelectrodes; Monitor; Mus; Nature; Neuromodulator; Neurons; Neurotransmitters; Parkinson Disease; Periodicity; Phase; Photometry; Play; Reaction Time; Regulation; Research; Research Personnel; Resolution; Role; Scanning; Signal Transduction; Slice; Stroke; Techniques; Technology; Testing; Therapeutic; Therapeutic Uses; Time; Traumatic Brain Injury; Universities; Virginia; Work; analytical tool; carbon fiber; design; in vivo; insight; model organism; neurochemistry; neuroprotection; neuroregulation; neurotransmission; new technology; real time monitoring; receptor; sensor; therapy development; tool

PROJECT SUMMARY What is the role of adenosine as a rapid modulator of neurotransmission and how can we harness its power for potential therapeutic use? To answer questions such as these, we need analytical tools that can measure multiple neurochemicals simultaneously with high temporal and spatial resolution. Our lab pioneered fast-scan cyclic voltammetry (FSCV) for adenosine, and discovered spontaneous, transient adenosine signaling that lasts only a few seconds. However, the range and effects of rapid adenosine neuromodulation are not well understood. Genetically-encoded sensors have recently been developed for neurotransmitter and calcium detection that offer high sensitivity, selectivity, and spatial resolution. While they can monitor a wide variety of neurochemicals, and not just electroactive molecules, there are still limited colors to detect different analytes. FSCV combined with genetically-encoded sensors would be advantageous to detect the neuromodulator adenosine and measure its downstream effects on dopamine and glutamate neurotransmission, as well as neuronal activity. The long-term goal of my lab is to develop new tools for monitoring real-time neuromodulation in the brain. The goal of this project is to develop multiplexed tools to understand neurochemical interactions and apply these tools to understand adenosine modulation of glutamate, dopamine, and calcium. The central hypothesis is that rapid adenosine release provides transient, but spatially localized, modulation of neurotransmitters in the brain. In the first Aim, we will develop multichannel FSCV, with an array of four electrodes, to determine how far adenosine diffuses in brain slices and the range of its neuromodulatory effects on dopamine release. In the second Aim, we will combine FSCV with genetically-encoded fluorescent sensors to probe the spatial and temporal profile of adenosine (measured with FSCV) modulation of dopamine (measured with GRABDA) or glutamate (measured with iGluSnFR). In the third Aim, we will combine multichannel FSCV and in vivo fiber photometry measurements of genetically-encoded sensors. We will demonstrate in vivo detection of adenosine, dopamine, and calcium changes to probe adenosine neuromodulation of neurotransmission and neuronal activity simultaneously. This research is significant because it develops tools that are broadly applicable for multiplexing neurotransmitter and neuromodulator measurements, harnessing the combined strengths of FSCV and genetically-encoded sensors. It is also significant because multiplexed tools will provide an unprecedented picture of the temporal and spatial dynamics of adenosine neuromodulation. The biological impact is understanding the rapid and local nature of adenosine neuromodulation, which is important for designing adenosine-based therapeutics for diseases such as Parkinson’s, ischemia, or traumatic brain injury where adenosine could be neuroprotective. The multiplexed tools could be applied to monitoring many other neurochemical interactions, in brain slices or in vivo, and will advance the field of neurochemical monitoring beyond one neurochemical at a time sensing.

项目概要 腺苷作为神经传递快速调节剂的作用是什么？我们如何利用它的力量 潜在的治疗用途？为了回答这些问题，我们需要能够测量的分析工具 我们的实验室开创了快速扫描技术，同时具有高时间和空间分辨率。 循环伏安法（FSCV）检测腺苷，并发现持续的自发、瞬时腺苷信号传导 然而，快速腺苷神经调节的范围和效果尚不清楚。 最近开发了用于神经递质和钙检测的基因编码传感器，可提供 高灵敏度、选择性和空间分辨率，同时它们可以监测多种神经化学物质，并且 不仅仅是电活性分子，用于检测不同分析物的 FSCV 颜色仍然有限。 基因编码传感器将有利于检测神经调节剂腺苷并测量其 对多巴胺和谷氨酸神经传递以及神经元活动的下游影响。 我实验室的目标是开发用于监测大脑实时神经调节的新工具。 该项目是开发多重工具来理解神经化学相互作用并将这些工具应用于 了解腺苷对谷氨酸、多巴胺和钙的调节作用的中心假设是快速。 腺苷释放对大脑中的神经递质提供短暂但空间局部的调节。 第一个目标是，我们将开发多通道 FSCV，具有四个电极阵列，以确定腺苷的距离 在第二个目标中，脑切片中的扩散及其对多巴胺释放的神经调节作用的范围。 我们将 FSCV 与基因编码荧光传感器结合起来，探测 腺苷（用 FSCV 测量） 多巴胺（用 GRABDA 测量）或谷氨酸（用 GRABDA 测量）的调节 在第三个目标中，我们将结合多通道 FSCV 和体内光纤光度测量。 我们将演示腺苷、多巴胺和钙的体内检测。 同时探测腺苷神经传递和神经元活动的变化。 研究意义重大，因为它开发了广泛适用于多路复用神经递质的工具 和神经调节剂测量，利用 FSCV 和基因编码的综合优势 这也很重要，因为多路复用工具将提供前所未有的时间图像。 腺苷神经调节的生物学影响是理解快速和局部的。 腺苷神经调节的性质，这对于设计基于腺苷的疗法非常重要 帕金森病、缺血或脑外伤等疾病，腺苷可以起到神经保护作用。 多重工具可用于监测脑切片或脑切片中的许多其他神经化学相互作用。 体内，并将推动神经化学监测领域超越一次传感一种神经化学物质。