Methods for Measuring Sub-second Neurotransmitter Interactions

测量亚秒级神经递质相互作用的方法

• 批准号: 8386454
• 负责人: Michael A. Johnson
• 金额: $ 21.96万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8386454
• 关键词: 4-hydroxyphenylacetic acid; AMPA Receptors; Address; Affect; Agonist; Amino Acid Neurotransmitters; Animals; Behavior; Biogenic Amines; Biological; Brain; Caliber; Cognitive; Corpus striatum structure; Detection; Development; Dopamine; Event; Excitatory Amino Acids; Exposure to; Fiber Optics; Generations; Gilles de la Tourette syndrome; Glutamates; Harvest; Huntington Disease; Hydrogen Peroxide; Impairment; Knowledge; Laboratories; Life; Light; Measurement; Measures; Mental Depression; Mercury; Methodology; Methods; Microelectrodes; Mission; Modeling; Movement Disorders; Mus; Nature; Nervous System Physiology; Neurobiology; Neuromodulator; Neurotransmitters; Nitric Oxide; Organism; Parkinson Disease; Pathway interactions; Physiologic pulse; Play; Rattus; Reaction; Regimen; Research; Research Personnel; Resolution; Role; Scanning; Serotonin; Signal Transduction; Site; Slice; System; Techniques; Technology; Testing; Time; Tissues; United States National Institutes of Health; Work; addiction; animal tissue; awake; burden of illness; carbon fiber; design; disability; empowered; gamma-Aminobutyric Acid; human disease; in vivo; innovation; insight; millisecond; neurochemistry; neurotransmitter release; photoactivation; receptor; research study; response; tool; uptake

DESCRIPTION (provided by applicant): The particular mechanisms underlying the sub-second control of dopamine release by glutamate and other neurotransmitters/neuromodulators are not well-known largely because the analytical methodology required to address these important questions has not been sufficiently developed. Although one technique, fast-scan cyclic voltammetry at carbon-fiber microelectrodes, offers sufficient temporal resolution to measure the release and uptake of electroactive biogenic amines such as dopamine, its use in characterizing such sub-second neurotransmitter/neuromodulator interactions is lacking. This deficiency represents a serious roadblock because signaling events in the brain that influence outward physical responses and cognitive events occur within this sub-second time regime. Therefore, the central aim of the proposed research is to develop and apply tools that will allow for the quantitative study of these neurotransmitter interactions in living tissues and in vivo. To temporally-compatible methods, fast-scan cyclic voltammetry and the photoactivation of the p- hydroxyphenyl caged form of glutamate (pHP-Glu), used here as a model caged system, will be applied to measure the sub-second response of dopamine release in response to millisecond timescale exposures to exogenous glutamate. The overall objective of this proposed research will be accomplished by successfully completing two specific aims: (1) Develop and validate an electrochemical method to quantitatively measure dopamine release and photoactivated glutamate simultaneously and (2) Optimize the construction and use of a combined photoactivation probe/carbon-fiber microelectrode for measuring the impact of caged compound photoactivation on dopamine release events in vivo. This approach is innovative because it is among the first to simultaneously apply these two high temporal resolution techniques in living tissues and animals. The development and application of this proposed methodology is significant because it will enable laboratory researchers to measure sub-second timescale dopamine release in response to ms-timescale glutamate application. Moreover, this work should have a broad impact since the application of these techniques can be expanded to include the detection of other electroactive neurotransmitters and neuromodulators, such as serotonin, hydrogen peroxide (H2O2), and nitric oxide (NO), and the photoactivation of other bioactive molecules, such as GABA and specifically designed synthetic agonists/antagonists. Importantly, this research directly relates to the NIH mission of seeking fundamental knowledge about the nature and behavior of living systems and reducing the burdens of illness and disability in that applies to, but is not limited to: (1) fundamental neurobiological studies; (2) studies of dopamine-related movement disorders [e.g. Huntington's disease (HD), Parkinson's disease (PD), Tourette's syndrome (TS)]; (3) studies of addiction and depression; and (4) studies addressing the mechanisms of action of CNS-active pharmacological agents. PUBLIC HEALTH RELEVANCE: This project aims at developing a method to study sub-second neurotransmitter interactions. These interactions likely play significant roles in the propagation of human disease. Therefore, a clearer understanding of these interactions has direct relevance to the NIH mission of seeking fundamental knowledge about the nature and behavior of living systems and reducing the burdens of illness and disability.

描述（由申请人提供）：谷氨酸和其他神经递质/神经调节剂对多巴胺释放的亚秒控制的基本机制并不是众所周知的，因为解决这些重要问题所需的分析方法尚未充分发展。 尽管一种技术是一种技术，即在碳纤维微电极上的快速扫描循环伏安法提供了足够的时间分辨率，可以测量诸如多巴胺等电活性生物胺的释放和吸收，其用于表征这种次级神经释放剂/神经递质/神经调节剂相互作用。 这种缺陷代表了一个严重的障碍，因为在这个次秒时，大脑中的信号事件会影响外向物理反应和认知事件。 因此，拟议的研究的核心目的是开发和应用工具，以允许对生物组织和体内这些神经递质相互作用进行定量研究。 To temporally-compatible methods, fast-scan cyclic voltammetry and the photoactivation of the p- hydroxyphenyl caged form of glutamate (pHP-Glu), used here as a model caged system, will be applied to measure the sub-second response of dopamine release in response to millisecond timescale exposures to exogenous glutamate. The overall objective of this proposed research will be accomplished by successfully completing two specific aims: (1) Develop and validate an electrochemical method to quantitatively measure dopamine release and photoactivated glutamate simultaneously and (2) Optimize the construction and use of a combined photoactivation probe/carbon-fiber microelectrode for measuring the impact of caged compound photoactivation on dopamine release events in vivo. 这种方法具有创新性，因为它是在生物组织和动物中同时同时应用这两种高时空技术的人之一。 这种提出的方​​法的开发和应用是重要的，因为它将使实验室研究人员能够根据MS-Timesscale谷氨酸的应用来测量下秒的多巴胺释放。 此外，这项工作应该具有广泛的影响，因为这些技术的应用可以扩展到包括对其他电活性神经递质和神经调节剂的检测，例如5-羟色胺过氧化氢（H2O2）和一氧化氮（NO）（no），以及其他生物活性分子的光活性，例如其他gaba和特定的gaba and gaba and gaba and gaba and gaba and gaba and toctife。 重要的是，这项研究直接涉及NIH的使命，即寻求有关生活系统的性质和行为的基本知识，并减轻适用的疾病和残疾负担，但不限于：（1）基本神经生物学研究； （2） 多巴胺相关运动障碍的研究[例如 亨廷顿氏病（HD），帕金森氏病（PD），图雷特综合症（TS）]； （3）成瘾和抑郁症的研究； （4）研究CNS活性药理剂作用机理的研究。 公共卫生相关性：该项目旨在开发一种研究次数神经递质相互作用的方法。 这些相互作用可能在人类疾病的传播中起着重要作用。 因此，对这些互动的更清晰的了解与NIH的使命是直接相关的，即寻求有关生命系统性质和行为并减轻疾病和残疾负担的基本知识。