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

描述（由申请人提供）：已知血清素神经递质系统调节情绪，焦虑状态和认知。此外，血清素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法分析方法提供的高时间和空间分辨率来表征与精神和退行性疾病直接相关的三种重要人类和小鼠模型中血清素神经传递的变化。本研究将：(1)利用高速时电流法评估人类淋巴母细胞培养物中血清素再摄取动力学的差异，这些培养物来源于两种常见的启动子多态性（5-HTTLPR和rs25531）和罕见的强迫症中发现的lle425Val编码区替代突变驱动的血清素转运体表达变化的个体；(2)采用快速循环伏安法表征血清素转运蛋白敲除小鼠和脑源性神经营养因子（BDNF）敲除小鼠体内血清素释放和再摄取的动态变化。我们的首要假设是，血清素能神经传递的潜在微妙但生物学上重要的变化发生在血清素转运体表达改变的小鼠和人类身上。此外，我们推测，在BDNF减少的小鼠中，血清素传递的变化是血清素能神经支配的年龄相关退行性丧失的基础。我们假设应用快速电化学方法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在多大程度上能够区分基因工程小鼠和具有遗传改变的人类细胞中改变的血清素神经传递，这对提高我们对精神和神经退行性疾病的发病机制和治疗的认识至关重要。

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.