Real-Time Detection of Glutamate using Templated Polymers as Shape-Changing Target Receptors

使用模板聚合物作为变形目标受体实时检测谷氨酸

• 批准号: 10195790
• 负责人: Edward Song
• 金额: $ 17.96万
• 依托单位: UNIVERSITY OF NEW HAMPSHIRE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10195790
• 关键词: Address; Adenosine; Affinity; Alzheimer' s Disease; Benchmarking; Binding; Binding Sites; Brain; Brain Diseases; Catecholamines; Chemicals; Communication; Complement; Data; Detection; Development; Diffusion; Dopamine; Ensure; Environment; Enzymes; Extracellular Space; Foundations; Frequencies; Future; Glutamate Receptor; Glutamates; Goals; Hippocampus (Brain); In Vitro; Label; Length; Longevity; Measurement; Measures; Mental disorders; Methods; Microdialysis; Microelectrodes; Modeling; Molecular; Molecular Conformation; Monitor; Morphology; Neurofibrillary Tangles; Neurons; Neurotransmitters; Outcome; Oxidation-Reduction; Parkinson Disease; Pathway interactions; Performance; Periodicity; Physiologic pulse; Physiological; Polymers; Process; Public Health; Reaction Time; Reporter; Research; Resolution; Sampling; Scanning; Serotonin; Shapes; Signal Transduction; Slice; Specificity; Spectrum Analysis; Stimulus; Synapses; Synaptic Cleft; System; Techniques; Technology; Time; Transducers; Validation; Work; aptamer; autism spectrum disorder; base; brain tissue; clinical application; copolymer; design; detection limit; electric impedance; extracellular; ferrocene; improved; in vivo; in vivo monitoring; liquid chromatography mass spectrometry; millisecond; monomer; mouse model; nervous system disorder; neurochemistry; novel; novel strategies; patch clamp; polymerization; real time monitoring; receptor; receptor binding; sensor; sensor technology; success; temporal measurement; uptake

PROJECT SUMMARY / ABSTRACT There is a great demand for a new sensing technology that can monitor neurochemicals in brain continuously in real-time with high temporal resolution. While electrochemical detection of electroactive neurotransmitters (such as dopamine and serotonin) have been successful using high frequency voltammetric methods (such as fast- scan cyclic voltammetry), measurement of non-electroactive species (such as glutamate) still remain a great challenge for achieving real-time monitoring with high time resolution. The goal of this project is to implement a new electrochemical sensing platform that can monitor glutamate, a well-known non-electroactive neurotransmitter, with physiologically relevant detection range and high temporal resolution. The proposed sensing approach can serve as an alternative to enzymatic sensing or microdialysis which have some limitations with respect to time resolution and stability. Motivated in part by the aptamer-based electrochemical sensors, the proposed sensing mechanism utilizes a novel synthetic target receptor as both a target recognition unit and a signal transducer. The glutamate receptor is formed by a single-chain stimuli-responsive templated polymer that binds specifically to its template molecule, namely, glutamate. Furthermore, upon selective target recognition, the polymer undergoes conformation change (from linear to folded shape). This change in polymer morphology can be electrochemically detected through the use of a redox reporter (such as ferrocene) attached to the polymer. For validation of the proposed sensing approach, the developed sensor will be compared against two well-established methods, a patch clamp and a microdialysis technique. A patch clamp system is ideal for measuring fast dynamics (milliseconds) of chemical exchange at the synaptic cleft of the neurons, however, lacks chemical specificity. A microdialysis in conjunction with liquid chromatography and mass spectrometry provides exceptional chemical specificity, however, the temporal resolution is poor (on the order of minutes). It is expected that the developed sensor platform will be able to bridge the gap between these two existing methods and to provide versatility in sensing performances. The goals of this project will be achieved by pursuing the following specific aims: (1) optimization of the templated polymer-based glutamate receptor to meet the desired performance metrics; and (2) validation of the developed glutamate sensor in a physiological environment. The successful outcome of this project will be the development of a new general platform technology for detection of neurochemicals in real-time with a time resolution that is sufficient for studying synaptic communications as well as for monitoring chemicals in the extracellular regions in the brain tissue.

项目概要/摘要 人们迫切需要一种能够连续监测大脑中神经化学物质的新型传感技术。 实时且具有高时间分辨率。虽然电化学检测电活性神经递质（例如 如多巴胺和血清素）已成功使用高频伏安法（例如快速伏安法） 扫描循环伏安法），非电活性物质（例如谷氨酸）的测量仍然很重要 实现高时间分辨率实时监控的挑战。该项目的目标是实施 新的电化学传感平台可以监测谷氨酸，一种众所周知的非电活性物质 神经递质，具有生理相关的检测范围和高时间分辨率。拟议的 传感方法可以作为酶传感或微透析的替代方法，但有一些局限性 关于时间分辨率和稳定性。部分受到基于适配体的电化学传感器的推动， 所提出的传感机制利用新型合成目标受体作为目标识别单元和 信号传感器。谷氨酸受体由单链刺激响应模板聚合物形成 特异性结合其模板分子，即谷氨酸。此外，根据选择性目标 识别后，聚合物会发生构象变化（从线性到折叠形状）。聚合物的这种变化 形态可以通过使用附加的氧化还原报告剂（例如二茂铁）进行电化学检测 到聚合物。为了验证所提出的传感方法，开发的传感器将与 两种行之有效的方法，膜片钳和微透析技术。膜片钳系统非常适合 然而，测量神经元突触间隙化学交换的快速动态（毫秒）， 缺乏化学特异性。微透析与液相色谱和质谱联用 提供了特殊的化学特异性，但是时间分辨率很差（大约分钟）。它 预计开发的传感器平台将能够弥补这两种现有方法之间的差距 并提供传感性能的多功能性。该项目的目标将通过追求 具体目标如下：（1）优化基于模板聚合物的谷氨酸受体以满足期望的 绩效指标； (2)在生理环境中验证所开发的谷氨酸传感器。这 该项目的成功成果将是开发一种新的通用平台技术来检测 实时神经化学物质的时间分辨率也足以研究突触通讯 至于监测脑组织细胞外区域的化学物质。