Monitoring Real-Time Neuropeptide Dynamics

监测实时神经肽动态

基本信息

批准号：
9383172
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
项目类别：
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/9383172
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 Technology Time Tissues United States National Institutes of Health addiction brain tissue burden of illness carbon fiber cost design drug of abuse drug reinforcement endogenous opioids evidence base experimental study extracellular hedonic improved in vivo innovation insight motivated behavior multi walled carbon nanotube nanoscale neurotransmission novel optogenetics rapid detection response reward processing sensor small molecule stressor temporal measurement therapeutic target tool

项目摘要

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.

抽象的：中脑边缘阿片肽在动机行为和奖励相关决策中的作用是尚不清楚，尽管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对这些分子释放的时间和地点的理解存在重大差距，因为缺乏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实时检测内源性脑啡肽（ENK）波动。该提案的目的是优化并充分表征该方法，以便提供社区拥有可用于研究 ENK 在复杂生理中的作用的既定工具从基本内分泌功能到动机的过程。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。我们还将调查分别对 20 种天然氨基酸中的每一种进行电化学分析，并在多变量中使用这些数据当这些残基被并入更长的氨基酸链。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），如以及有前景的纳米电极材料。最后，第三个目标是评估实时ENK 大鼠肾上腺和脑组织的动态，以便深入了解正常浓度范围， ENK 的细胞外寿命和扩散特征（影响范围），以及生理和可以诱导肽能信号变化的药理学条件。我们也会直接比较使用 FSCV 研究小分子递质和 ENK 的刺激-反应关系。这个项目是这是实现我们长期目标的关键一步，即阐明释放和清除动态几种神经肽和小分子是动机行为的不同方面的基础。它将澄清有关内源性阿片肽信号传导基本性质的悬而未决的问题，以及使 FSCV 能够被自信地使用来揭示关键的机制细节，从而为基于证据的信息提供信息用于治疗多种疾病的药物疗法，包括药物滥用疾病。