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: 10365275
• 负责人: B. JILL VENTON
• 金额: $ 62.94万
• 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/10365275
• 关键词: Acetylcholine; Adenosine; Animal Model; Biological; Brain; Calcium; Calcium Signaling; Cells; Color; Detection; Development; Diffuse; 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; base; carbon fiber; design; in vivo; insight; 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.

项目摘要 腺苷作为神经传递的快速调节剂的作用是什么？ 潜在的治疗用途？为了回答这些问题，我们需要分析工具， 同时具有高时间和空间分辨率的多种神经化学物质。我们的实验室开创了快速扫描 循环伏安法（FSCV）的腺苷，并发现自发的，短暂的腺苷信号，持续 只有几秒钟。然而，腺苷快速神经调节的范围和作用还不清楚。 最近已经开发出用于神经递质和钙检测的遗传编码传感器， 高灵敏度、选择性和空间分辨率。虽然他们可以监测各种各样的神经化学物质， 不仅仅是电活性分子，仍然存在有限的颜色来检测不同的分析物。FSCV结合 基因编码的传感器将有利于检测神经调节剂腺苷并测量其浓度。 对多巴胺和谷氨酸神经传递以及神经元活动的下游影响。长期 我实验室的目标是开发新的工具来监测大脑中的实时神经调节。这个目标 一个项目是开发多路复用的工具来了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。核心假设是， 腺苷释放提供脑中神经递质的瞬时但空间定位的调节。在 第一个目标，我们将开发多通道FSCV，具有四个电极的阵列，以确定腺苷 在脑切片中扩散，以及其对多巴胺释放的神经调节作用范围。在第二个目标中， 我们将联合收割机FSCV与基因编码的荧光传感器相结合， 腺苷（用FSCV测量）调节多巴胺（用GRABDA测量）或谷氨酸（用GABDA测量） iGluSnFR）。在第三个目标中，我们将结合联合收割机多通道FSCV和在体纤维光度测量 基因编码的传感器。我们将演示腺苷、多巴胺和钙的体内检测 改变以同时探测神经传递和神经元活动的腺苷神经调节。这 这项研究意义重大，因为它开发了广泛适用于多路神经递质的工具， 和神经调质测量，利用FSCV和基因编码的 传感器.这也是重要的，因为多路复用工具将提供一个前所未有的图片的时间 和腺苷神经调节的空间动力学。生物学的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治疗剂是重要的， 例如帕金森氏症、局部缺血或创伤性脑损伤等疾病，其中腺苷可以起到神经保护作用。 多路复用工具可以应用于监测许多其他神经化学相互作用，在大脑切片或 体内，并将推进神经化学监测领域超越一次一种神经化学物质的传感。