Direct, Real-Time Monitoring of Gliotransmitter Release

描述（由申请人提供）：本提案旨在开发一种纳米级电化学传感器，用于确定胶质递质在中枢神经系统中作为细胞间信号分子的作用。传感平台将结合电化学提供的化学特异性，基于适配体的传感器与纳米级电极提供的精致时空分辨率，并将适用于膜片钳仪器。这项提议的目的有两个。首先，纳米级电化学适配体传感器的制造、表征和优化将导致能够特异性监测星形胶质细胞中三磷酸腺苷（ATP）释放的高性能传感器。其次，传感器将用于确定ATP释放的主要机制，以及ATP释放和机制是否局限于星形胶质细胞的特定区域。本提案的最初重点将是利用纳米传感器对培养的星形胶质细胞进行体外测量的发展。然而，由于传感器在复杂的生物介质中发挥作用，从长远来看，它们将被应用于更多的生理模型（脑切片），并最终优化用于体内，以阐明ATP作为信号分子的作用。这项研究的长期目标是了解神经胶质细胞通讯的分子基础，以加速发现和减轻神经系统疾病的负担。提出的研究将产生一种尖端的工具，能够通过提供理解中枢神经系统功能的方法来改变神经胶质细胞的研究。

DESCRIPTION (provided by applicant): This proposal aims to develop a nanoscale electrochemical sensor that will be used to determine the role gliotransmitters play as intercellular signaling molecules in the central nervous system. The sensing platform will combine the chemical specificity afforded by electrochemical, aptamer-based sensors with the exquisite spatiotemporal resolution afforded by nanoscale electrodes and will be amenable to use with patch clamp instrumentation. The aims of the proposal are two-fold. First, the fabrication, characterization and optimization of nanoscale, electrochemical aptamer-based sensors will lead to high-performance sensors capable of specifically monitoring adenosine triphosphate (ATP) release from astrocytes. Second, sensors will be used to determine the dominant mechanism(s) of ATP release and whether ATP release and mechanisms are localized to specific regions of the astrocyte. The initial focus in this proposal will be the development of in vitro measurements with the nanosensors on cultured astrocytes. However, because the sensors developed function in complex biological media, they will, in the longer term, be applied to more physiological models (brain slices) and ultimately optimized for use in vivo to elucidate the role of ATP as a signaling molecule. The long-term goal of the research is to understand the molecular basis of glial cell communication in order to accelerate discoveries and reduce the burden of nervous system disorders. The proposed studies will yield a cutting edge tool that that is capable of transforming glial cell research by providing methodology to understand function in the central nervous system.