Developing Electrochemical Sensors to Enable Quantitative Measure of Gliotransmitter Release from Astrocytes

开发电化学传感器以定量测量星形胶质细胞释放的胶质递质

• 批准号: 10752836
• 负责人: Ryan J. White
• 金额: $ 39.23万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752836
• 关键词: Affect; Affinity; Age; Architecture; Astrocytes; Binding; Biological; Brain; Brain region; Cell surface; Cells; Chemicals; Chemistry; Collagen; Communication; Corpus striatum structure; DNA; Detection; Development; Diameter; Disease; Electrochemistry; Electrodes; Environment; Exocytosis; Frequencies; Glutamates; Goals; Heterogeneity; Hippocampus; Homeostasis; Hot Spot; Immobilization; Ionomycin; Knowledge; Left; Libraries; Maps; Measurement; Measures; Molecular; Molecular Conformation; Monitor; Neurodegenerative Disorders; Neuroglia; Neurons; Nucleic Acids; Organelles; Physiologic pulse; Physiological; Population; Process; Proteins; Research; Role; Science; Serine; Signal Transduction; Specificity; Structure; Surface; System; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Transcriptional Regulation; aptamer; brain dysfunction; cell type; design; developmental disease; gamma-Aminobutyric Acid; innovation; insight; meter; molecular dynamics; nano; nanomolar; nanoscale; nervous system disorder; neuronal cell body; neurovascular; real time monitoring; response; response to injury; sensor; spatiotemporal; synaptic function; technology development; temporal measurement; three dimensional cell culture; tool; transmission process

PROJECT SUMMARY Objectives. The primary objective of this proposal is to develop chemically specific electrochemical sensors to provide rapid and direct measurements of physiologically relevant chemical messenger release (technology development). The secondary objective is to apply these sensors to determine signaling heterogeneity in astrocytes from different brain regions (biological hypothesis). Significance and Knowledge Gap. Astrocytes act as integrators across many circuits and environments in the brain and how they interact with other cell types can vary in time and space. Rigorous prior research demonstrates that astrocytes are heterogenous, varying by brain region and circuit. Heterogeneity can be affected by transcriptional control related to synapse function, plasticity, molecular transmission and protein machinery and organelles that underly gliotransmission. Astrocyte heterogeneity also affects how this class of cells respond to insults and age and can potentially be predictive of disease vulnerability. Astrocytes have typically been studied as a homogenous population, thus there is a need to study functional heterogeneity of astrocytes (e.g., signaling) to determine functional roles of astrocytes in development, response to injuries, and neurodegenerative disease and how this response influences local circuit function and homeostasis. Solution and Specific Aims. A major barrier to studying gliotransmission is the lack of measurement tools that possess the combined spatiotemporal and chemical specificity to study dynamic molecular signaling from astrocytes. We aim to develop a sensor platform that overcomes this barrier by providing direct and rapid measurement of gliotransmission over broad spatial and temporal ranges. Through a collaborative proposal we propose to leverage the universal and specific chemical detection abilities of electrochemical, aptamer-based (E-AB) sensors with innovative measurement science for rapid determination of gliotransmitter dynamics and heterogeneity. With this new measurement technology, we aim to test the hypothesis that gliotransmitter signaling varies in terms of the frequency, amount, and identity of transmitters released with circuit-, and inter- and intraregional specificity. We will 1) Develop electrochemical, aptamer-based (E-AB) sensors to monitor real-time release of gliotransmitters from a cell population in 3D culture. 2) Develop recessed, microscale E-AB sensors to monitor the release of gliotransmitters from single astrocytes. 3) Develop recessed, nanoscale sensors to monitor the release of gliotransmitters from sub-cellular regions. 4) Select and characterize highly specific structure-switching nucleic acid aptamers as binding partners for glutamate, GABA, and D-serine with nM affinity for use in aims 1-3.

项目摘要 目标。该建议的主要目的是开发化学特定的电化学传感器以提供 快速和直接测量生理相关的化学信使释放（技术开发）。这 次要目标是应用这些传感器来确定来自不同大脑区域的星形胶质细胞中的信号异质性 （生物学假设）。 意义和知识差距。星形胶质细胞充当大脑中许多电路和环境中的集成商 它们与其他单元类型的相互作用如何在时间和空间上有所不同。严格的先前研究表明，星形胶质细胞是 异源，随大脑区域和电路而变化。异质性可能会受到与突触有关的转录控制的影响 功能，可塑性，分子传递和蛋白质机制以及基础旋转传递的细胞器。星形胶质细胞 异质性还会影响这类细胞对侮辱和年龄的反应，并有可能预测疾病 脆弱性。星形胶质细胞通常被研究为同质种群，因此需要研究功能 星形胶质细胞（例如信号传导）的异质性，以确定星形胶质细胞在发育中的功能作用，对伤害的反应， 神经退行性疾病以及该反应如何影响局部回路功能和稳态。 解决方案和特定目的。研究Gliotsimers的主要障碍是缺乏具有的测量工具 合并的时空特异性和化学特异性，用于研究星形胶质细胞的动态分子信号传导。我们旨在发展 一个传感器平台，通过提供直接和快速测量Gliotsermission来克服这一障碍的传感器平台 空间和时间范围。通过协作提案，我们建议利用通用和特定的化学物质 具有创新测量科学的电化学，基于适体的（E-AB）传感器的检测能力 神经闪光灯动力学和异质性的测定。借助这种新的测量技术，我们旨在测试 假设闪光灯闪光灯信号的信号随释放发射机的频率，数量和身份而变化 电路和区域内和区域内特异性。我们将1）开发电化学，基于适体的（E-AB）传感器 监测3D培养中细胞种群中胶质递质的实时释放。 2）发展凹陷，微观e-ab 传感器以监测单个星形胶质细胞中闪光灯的释放。 3）开发嵌入式，纳米级传感器以监测 从亚细胞区域释放胶质递质。 4）选择并表征高度特定的结构开关 核酸适体作为谷氨酸，GABA和D丝氨酸的结合伴侣，具有NM亲和力，可用于AIMS 1-3。