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/神经调节剂亚秒控制多巴胺释放的具体机制并不为人所知，很大程度上是因为解决这些重要问题所需的分析方法尚未得到充分发展。 尽管一种技术，即碳纤维微电极的快速扫描循环伏安法，提供了足够的时间分辨率来测量多巴胺等电活性生物胺的释放和吸收，但其在表征这种亚秒级神经递质/神经调节剂相互作用方面的应用尚缺乏。 这种缺陷代表了一个严重的障碍，因为影响外在身体反应和认知事件的大脑信号事件发生在这个亚秒级的时间范围内。 因此，拟议研究的中心目标是开发和应用能够定量研究活组织和体内这些神经递质相互作用的工具。 对于时间兼容的方法，快速扫描循环伏安法和对羟基苯基笼形谷氨酸（pHP-Glu）的光活化（此处用作模型笼式系统）将用于测量多巴胺释放的亚秒响应响应于毫秒时间尺度的外源谷氨酸暴露。 这项研究的总体目标将通过成功完成两个具体目标来实现：（1）开发和验证一种电化学方法来同时定量测量多巴胺释放和光活化谷氨酸；（2）优化组合光活化探针/的构建和使用碳纤维微电极用于测量笼状化合物光活化对体内多巴胺释放事件的影响。 这种方法具有创新性，因为它是第一个在活体组织和动物中同时应用这两种高时间分辨率技术的方法。 这种提出的方​​法的开发和应用具有重要意义，因为它将使实验室研究人员能够测量响应于毫秒时间尺度谷氨酸应用的亚秒时间尺度多巴胺释放。 此外，这项工作应该会产生广泛的影响，因为这些技术的应用可以扩展到包括其他电活性神经递质和神经调节剂的检测，例如血清素、过氧化氢（H2O2）和一氧化氮（NO），以及光激活其他生物活性分子，例如 GABA 和专门设计的合成激动剂/拮抗剂。 重要的是，这项研究与 NIH 的使命直接相关，即寻求有关生命系统的性质和行为的基础知识，并减轻疾病和残疾的负担，适用于但不限于：（1）基础神经生物学研究； (2) 多巴胺相关运动障碍的研究[例如 亨廷顿病（HD）、帕金森病（PD）、抽动秽语综合征（TS）]； (3)成瘾和抑郁症的研究； (4) 研究中枢神经系统活性药物的作用机制。 公共健康相关性：该项目旨在开发一种研究亚秒级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发挥重要作用。 因此，更清楚地了解这些相互作用与 NIH 的使命直接相关，即寻求有关生命系统的性质和行为的基础知识以及减轻疾病和残疾的负担。