Serotonin Transporter Kinetics In Vivo by Microdialysis/Capillary UPLC

通过微透析/毛细管 UPLC 测定体内血清素转运蛋白动力学

• 批准号: 7599178
• 负责人: STEPHEN G. WEBER
• 金额: $ 19.19万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7599178
• 关键词: Animals; Antidepressive Agents; Anxiety; Appetitive Behavior; Behavior; Biological; Blood capillaries; Brain; Cell membrane; Chemistry; Cognition; Communication; Complex; Coupled; Detection; Disease; Electron Transport; Extracellular Space; Genetic Variation; Goals; High Pressure Liquid Chromatography; High temperature of physical object; Human; Kinetics; Measurement; Measures; Methodology; Methods; Microdialysis; Moods; Motor Activity; Mus; Nervous System Physiology; Neuraxis; Neurotransmitters; Organism; Oxidation-Reduction; Peptides; Personality Traits; Pharmaceutical Preparations; Play; Population; Predisposition; Psychopathology; Psychophysiology; Rewards; Role; Sampling; Selective Serotonin Reuptake Inhibitor; Serotonin; Signal Transduction; Speed; Stress; System; Therapeutic; Time; analytical method; awake; capillary; capillary liquid chromatography; clinically significant; depression; extracellular; in vivo; meetings; monoamine; neurotransmission; novel; pressure; presynaptic; public health relevance; response; reuptake; serotonin transporter; uptake

DESCRIPTION (provided by applicant): We propose to develop an analytical method using capillary high-performance liquid chromatography at high temperature and pressure coupled to photoluminescence detection following electron transfer (UPLC-PFET) following online microdialysis sampling to enable fast serotonin neurotransmitter measurements in the brains of awake behaving animals. This method will provide substantial improvements in the speed and limits of detection over current methods (e.g., HPLC-EC, CE-LIF). It will be broadly applicable to other monoamine neurotransmitters and metabolites and peptides. The brain serotonin neurotransmitter system regulates many important psychophysiological functions including mood, anxiety states, cognition, reward-related behavior, motor activity and appetitive behavior. The presynaptic plasma membrane serotonin transporter (SERT) takes up serotonin from the extracellular space and plays a central role in regulating serotonergic neurotransmission. It is the primary target for the most widely prescribed class of antidepressant and anti-anxiety medications } the serotonin reuptake inhibitors (SRIs). In humans, reduced SERT expression driven by a commonly occurring gene variant has been associated with increases in anxiety-related personality traits. and susceptibility to stress-associated depression. To investigate effects of decreased SERT, we have produced mice that express reduced SERT. Decreased SERT in mice results in increased anxiety-like behavior similar to that observed in humans. Despite the key role that serotonin uptake plays, the kinetics of the uptake of endogenous serotonin has never been measured We will employ the optimized UPLC-PFET method developed here to measure these kinetics. PUBLIC HEALTH RELEVANCE: To understand the brain's internal communications, we need analytical methods that are absolutely selective and also fast. There are no general approaches that have both attributes. We will develop one, and apply it to a question about serotonin - the key player in anxiety, depression and related disorders.

描述（由申请人提供）：我们建议在高温下使用毛细血管高性能液相色谱法开发分析方法，并在电子传输后与光致发光检测结合的压力在线微透析采样后，以启用快速的血清素神经释放剂测量，以启用快速的血清素神经释放剂测量。此方法将对当前方法（例如HPLC-EC，CE-LIF）的检测速度和限制进行实质性改善。它将广泛适用于其他单胺神经递质，代谢产物和肽。脑血清素神经递质系统调节许多重要的心理生理功能，包括情绪，焦虑状态，认知，与奖励相关的行为，运动活动和食用性行为。突触前质膜血清素转运蛋白转运蛋白（SERT）从细胞外空间中占据5-羟色胺，并在调节血清素能神经传递方面起着核心作用。它是最广泛规定的抗抑郁药和抗焦虑药的主要目标}羟色胺再摄取抑制剂（SRIS）。在人类中，由常见发生的基因变异驱动的SERT表达降低与焦虑相关人格特征的增加有关。以及对压力相关抑郁症的敏感性。为了研究SERT降低的影响，我们产生了表达降低SERT的小鼠。小鼠的SERT减少导致焦虑样行为的增加，类似于人类观察到的行为。尽管5-羟色胺摄取作用的关键作用，但从未测量过内源5-羟色胺的动力学，我们将采用此处开发的优化的UPLC-PFET方法来测量这些动力学。 公共卫生相关性：要了解大脑的内部通信，我们需要绝对选择性且快速的分析方法。没有两个属性的一般方法。我们将开发一个，并将其应用于有关5-羟色胺的问题 - 焦虑，抑郁症和相关疾病的关键人物。

项目成果

Capillary ultrahigh performance liquid chromatography with elevated temperature for sub-one minute separations of basal serotonin in submicroliter brain microdialysate samples.

• DOI: 10.1021/ac102200q
• 发表时间: 2010-12-01
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Liu, Yansheng;Zhang, Jing;Xu, Xiaomi;Zhao, Moe K.;Andrews, Anne M.;Weber, Stephen G.
• 通讯作者: Weber, Stephen G.