Voltammetry of 5-HT Transmission in Psychiatric & Degenerative Disease Models

描述(申请人提供)：已知的5-羟色胺神经递质系统调节情绪、焦虑状态和认知。此外，5-羟色胺传递的改变被认为与情绪和焦虑障碍以及包括阿尔茨海默病在内的神经退行性疾病的病因和治疗有关。该项目旨在利用碳纤维微电极伏安分析方法提供的高时间和空间分辨率来表征与精神疾病和退行性疾病直接相关的三个重要人类和小鼠模型中5-羟色胺神经传递的变化。拟议的研究将：(1)使用高速计时电流法，评估由两个常见启动子多态(5-HTTLPR和rs25531)以及在罕见常见形式的强迫症中发现的lle425Val编码区替代突变驱动的可变5-羟色胺转运体表达的人淋巴母细胞培养物5-羟色胺再摄取动力学的差异；(2)使用快速循环伏安法研究5-羟色胺转运体基因敲除小鼠和脑源性神经营养因子(BDNF)基因敲除小鼠体内5-羟色胺释放和重摄取动力学的变化。我们的主要假设是，在5-羟色胺转运体表达改变的小鼠和人类中，5-羟色胺能神经传递发生了潜在的细微但具有生物学意义的变化。此外，我们的理论认为，5-羟色胺传递的变化是BDNF减少的小鼠5-羟色胺能神经与年龄相关的退行性丢失的原因。我们推测，应用快速电化学方法来检测脑神经传递的这些变化是必要的，这是研究基础和疾病相关过程的基础。这些研究的结果将揭示伏安技术能够在多大程度上区分具有基因改变的基因工程小鼠和人类细胞中改变的5-羟色胺神经传递，这些改变对于增进我们对精神和神经退行性疾病的发病机制和治疗的了解非常重要。

DESCRIPTION (provided by applicant): The serotonin neurotransmitter system is known to regulate emotion, anxiety states and cognition. Moreover, altered serotonin transmission is hypothesized to be involved in the etiology and treatment of mood and anxiety disorders, and neurodegenerative diseases including Alzheimer's disease. This project is designed to take advantage of the high temporal and spatial resolution afforded by carbon fiber microelectrode voltammetry analytical methods to characterize changes in serotonin neurotransmission in three important human and mouse models with direct relevance to psychiatric and degenerative disorders. The proposed research will: (1) Use high-speed chronoamperometry to evaluate differences in the kinetics of serotonin reuptake in human lymphoblast cell cultures derived from individuals with variable serotonin transporter expression driven by two common promoter polymorphisms (5-HTTLPR and rs25531) in combination with an lle425Val coding region substitution mutation found in rare familiar forms of obsessive compulsive disorder; and (2) Employ fast cyclic voltammetry to characterize alterations in the dynamics of serotonin release and reuptake in vivo in serotonin transporter knockout mice and brain-derived neurotrophic factor (BDNF) knockout mice. Our overarching hypothesis states that potentially subtle but biologically important changes in serotonergic neurotransmission occur in mice and humans with altered serotonin transporter expression. Further, we theorize that changes in serotonin transmission underlie age-related degenerative loss of serotonergic innervation in mice with reduced BDNF. We postulate that application of fast electrochemical methods is necessary to detect these changes in brain neurotransmission, which are fundamental to the investigation of basic and disease-related processes. This outcome of these studies will reveal the extent to which voltammetric techniques are able to differentiate altered serotonin neurotransmission in genetically engineered mice and human cells with genetic alterations important for advancing our knowledge of the pathogenesis and treatment of psychiatric and neurodegenerative diseases.