Serotonin Transporter Kinetics In Vivo by Microdialysis/Capillary UPLC

描述（由申请人提供）：我们提议开发一种分析方法，该方法在高温高压下使用毛细管高效液相色谱法，在在线微透析采样后结合电子转移后光致发光检测（UPLC-PFET），以快速测量清醒行为动物大脑中的5-羟色胺神经递质。该方法将在速度和检测极限方面提供比当前方法（例如，HPLC-EC，CE-LIF）。它将广泛适用于其他单胺神经递质和代谢物和肽。脑5-羟色胺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、奖励相关行为、运动活动和食欲行为。突触前质膜5-羟色胺转运体（SERT）从细胞外空间摄取5-羟色胺，并在调节多巴胺能神经传递中起核心作用。它是最广泛处方的抗抑郁和抗焦虑类药物血清素再摄取抑制剂（SRI）的主要靶点。在人类中，由一种常见的基因变异驱动的SERT表达减少与焦虑相关的人格特征增加有关。和易受压力相关抑郁症的影响。为了研究减少的SERT的影响，我们已经产生了表达减少的SERT的小鼠。在小鼠中SERT降低导致类似于在人类中观察到的焦虑样行为增加。尽管5-羟色胺摄取起着关键作用，但内源性5-羟色胺摄取的动力学从未被测量过。我们将采用本文开发的优化的UPLC-PFET方法来测量这些动力学。 公共卫生相关性：为了了解大脑的内部通信，我们需要绝对有选择性且快速的分析方法。不存在具有这两种属性的通用方法。我们将开发一个，并将其应用于一个关于血清素的问题-焦虑，抑郁和相关疾病的关键球员。

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.