Monitoring Real-Time Neuropeptide Dynamics

监测实时神经肽动态

• 批准号: 9978009
• 负责人: LESLIE A SOMBERS
• 金额: $ 37.89万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978009
• 关键词: Adrenal Glands; Amino Acids; Behavior; Biological; Biological Models; Brain; Catecholamines; Chemical Stimulation; Chemicals; Chromaffin Cells; Collaborations; Communities; Complex; Corpus striatum structure; Data; Decision Making; Detection; Development; Diffusion; Discrimination; Disease; Dopamine; Drug Addiction; Electrochemistry; Electrodes; Endocrine; Engineering; Enkephalins; Extracellular Space; Frequencies; Goals; Health; In Situ; Individual; Knowledge; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Membrane; Methodology; Methods; Microdialysis; Microelectrodes; Mission; Monitor; Motivation; Nafion; Nature; Neuropeptides; Neurotransmitters; Opioid; Opioid Peptide; Peptide Hydrolases; Peptide Signal Sequences; Peptides; Periodicity; Pharmacology; Pharmacotherapy; Physiologic pulse; Physiological; Physiological Processes; Play; Public Health; Rattus; Research; Resolution; Rewards; Role; Sampling; Scanning; Signal Transduction; Slice; Substance abuse problem; Surface; System; Systems Development; Time; Tissues; United States National Institutes of Health; addiction; brain tissue; burden of illness; carbon fiber; cost; design; double walled carbon nanotube; drug of abuse; drug reinforcement; endogenous opioids; evidence base; experimental study; extracellular; hedonic; improved; in vivo; individual response; innovation; insight; motivated behavior; nanoscale; neurotransmission; novel; optogenetics; rapid detection; real time monitoring; response; reward processing; sensor; sensor technology; small molecule; stressor; temporal measurement; therapeutic target; tool

ABSTRACT: The role of mesolimbic opioid peptides in motivated behavior and reward-related decision making is unclear, despite extensive evidence indicating that these molecules are important mediators of hedonic and motivational aspects of reward processing, and the fundamental response to drugs of abuse. This is largely due to a critical gap in understanding when and where these molecules are released, because there is a paucity of detection methods for monitoring opioid peptides in the extracellular space. We have established the feasibility of using fast-scan cyclic voltammetry (FSCV) and carbon- fiber microelectrodes in tissue for the detection of endogenous enkephalin (ENK) fluctuations in real time. The objective of this proposal is to optimize and fully characterize this methodology, so as to provide the community with an established tool that can be used to study the role of the ENKs in complex physiological processes ranging from basic endocrine function to motivation. The first goal is to fully characterize selectivity. We will assess incorporation of well-characterized Nafion composite membranes into the sensor design as a physical means to enhance selective detection. We will also investigate the electrochemistry of each of 20 natural amino acids individually, and use these data in a multivariate approach to identify individual amino acid contributors to the voltammetric signal when these residues are incorporated into longer amino acid chains. The second goal is to optimize sensitivity by systematically investigating electrochemical parameters (scan rates, holding potentials, and sampling frequencies), as well as promising nanoscale electrode materials. Finally, the third goal is to evaluate real-time ENK dynamics in rat adrenal and brain tissue, so as to provide insight into the normal concentration range, extracellular lifetime, and diffusion profile (sphere of influence) of the ENKs, as well as physiological and pharmacological conditions that can induce changes in peptidergic signaling. We will also directly compare the stimulation-response relationship for small molecule transmitters and ENK, using FSCV. This project is a critical step toward our long-term goal of elucidating how the release and clearance dynamics of several neuropeptides and small molecules underlie discrete aspects of motivated behavior. It will clarify outstanding questions regarding the fundamental nature of endogenous opioid peptide signaling, and enable FSCV to be used confidently to reveal critical mechanistic details that will inform evidence-based pharmacotherapies for treating a wide range of disorders, including substance abuse disorders.

摘要： 中脑边缘阿片肽在动机行为和奖励相关决策中的作用是 尽管有大量证据表明这些分子是享乐的重要介质， 奖励处理的动机方面，以及对滥用药物的基本反应。这 很大程度上是由于在理解这些分子何时何地被释放方面存在关键差距， 由于缺乏用于监测细胞外阿片肽的检测方法 空间我们已经建立了使用快速扫描循环伏安法（FSCV）和碳- 组织中的纤维微电极用于真实的检测内源性脑啡肽（ENK）波动。 本提案的目的是优化和充分说明这一方法，以便提供 社区与一个既定的工具，可用于研究的作用，ENK在复杂的生理 从基本的内分泌功能到动机。第一个目标是充分表征 选择性我们将评估将充分表征的Nafion复合膜纳入 传感器设计作为增强选择性检测的物理手段。我们还将调查 每个20天然氨基酸的电化学单独，并使用这些数据在多变量 当这些残基被还原时， 被并入更长的氨基酸链中。第二个目标是通过系统地优化灵敏度， 研究电化学参数（扫描速率、保持电位和采样频率）， 也是很有前途的纳米电极材料。最后，第三个目标是评估实时ENK 大鼠肾上腺和脑组织中的动力学，以便深入了解正常浓度范围， 细胞外寿命和ENK的扩散分布（影响范围），以及生理和 药理学条件可以诱导肽能信号传导的变化。我们也会直接比较 小分子递质和ENK的刺激-反应关系，使用FSCV。这个项目是 这是朝着我们的长期目标迈出的关键一步， 几种神经肽和小分子是动机行为的各个方面的基础。它将 澄清有关内源性阿片肽信号传导的基本性质的悬而未决的问题， 使FSCV能够自信地用于揭示关键的机制细节， 药物疗法用于治疗广泛的病症，包括物质滥用病症。