A Scalable N x N Electrochemical Detector Array Platform for Analysis of Quantal

用于量子分析的可扩展 N x N 电化学检测器阵列平台

• 批准号: 8322641
• 负责人: Manfred LINDAU
• 金额: $ 37.06万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8322641
• 关键词: Adrenal Glands; Affect; Amplifiers; Binding; Biological Assay; Botox; Brain; Catecholamines; Cell Communication; Cell Count; Cell Size; Cells; ChIP-on-chip; Characteristics; Chromaffin Cells; Cosmetics; Cytoplasmic Granules; Deposition; Development; Dopamine; Electrodes; Event; Exocytosis; Extracellular Space; Frequencies; Goals; Hormones; Individual; Industry; Kinetics; Levodopa; Measurement; Measures; Medical; Membrane; Methods; Microelectrodes; Microscopic; Modification; Molecular; Names; Neurobiology; Neurons; Neurotransmitters; Noise; Norepinephrine; Organelles; PC12 Cells; Parkinson Disease; Performance; Pharmaceutical Preparations; Positioning Attribute; Process; Property; Proteins; Regulation; Research; Resolution; Semiconductors; Series; Serotonin; Signal Transduction; Signaling Molecule; Solutions; Surface; Technology; Testing; Time; Vesicle; Work; base; carbon fiber; detector; dopaminergic neuron; drug discovery; interest; meetings; neurotransmitter release; novel; response; sensor

DESCRIPTION (provided by applicant): The goal of this project is the development of a scalable n x n electrochemical detector array platform with on- chip amplifiers for massively parallel recordings of quantal transmitter release events. The neurobiological process that this assay will analyze is the process of exocytosis and transmitter release, one of the key processes in brain function and beyond. The molecules to be released are stored at high concentration in membrane-bound organelles. Upon stimulation the contents of these vesicles are released in quantal events through a fusion pore that connects the vesicular lumen to the extracellular space. Understanding the mechanisms of vesicle fusion and transmitter release is of broad medical significance. In the treatment of Parkinson's disease the drug levodopa increases dopamine release from the reduced number of dopaminergic neurons. On the other hand, BoTox treatment exerts its effect through the reduction of transmitter release. In addition to these examples, many other drugs and many molecular manipulations modulate transmitter release in various ways. This regulation of transmitter release occurs not only via changing the number or frequency of quantal release events but also via modulation of quantal size and of the kinetics of release from individual vesicles. To understand the mechanism by which a specific manipulation affects transmitter release it is therefore necessary to perform precise measurements of individual quantal release events, analyze their amplitude and time course and derive characteristic parameters. Conventionally, such measurements are performed by positioning a carbon fiber microelectrode close to a cell (such as a chromaffin cell, a dopaminergic or serotonergic neuron or a PC12 cell) under microscopic observation, stimulate the cell and record a series of release events. The technology developed in this project is adapted from the semiconductor industry and involves the development of a CMOS microelectronic chip for on-chip recordings of single quantal release events of oxidizable transmitter molecules such as noradrenaline, dopamine, or serotonin. The technology will allow the simultaneous recording of single vesicle release events from hundreds of cells without the need for microscopic observation and manipulation, and will thereby provide a high-throughput platform to characterize molecular and pharmacological manipulations. The technology will accelerate the research aimed at understanding the molecular mechanisms of transmitter release and its modulation as well as the development and testing of treatments that act through the modulation of transmitter release. In this way this novel assay platform would provide opportunities to measure quantal release events as neurobiological endpoints and will provide a pipeline for target identification and drug discovery.

描述（由申请人提供）：该项目的目标是开发一个可扩展的n × n电化学探测器阵列平台，该平台带有片上放大器，用于大规模并行记录量子发射器释放事件。本实验将分析的神经生物学过程是胞吐和递质释放的过程，这是大脑功能及其他方面的关键过程之一。被释放的分子以高浓度储存在膜结合细胞器中。刺激后，这些囊泡的内容物通过连接囊泡腔和细胞外空间的融合孔以量子方式释放。了解囊泡融合和递质释放的机制具有广泛的医学意义。在帕金森氏症的治疗中，药物左旋多巴增加了多巴胺能神经元数量减少的多巴胺释放。另一方面，肉毒杆菌素治疗通过减少递质释放来发挥作用。除了这些例子，许多其他药物和许多分子操作以各种方式调节递质释放。这种对递质释放的调节不仅通过改变量子释放事件的数量或频率发生，而且还通过对量子大小和单个囊泡释放动力学的调节发生。为了理解特定操作影响发射器释放的机制，因此有必要对单个量子释放事件进行精确测量，分析其幅度和时间过程并推导特征参数。通常，这种测量是通过将碳纤维微电极靠近细胞（如染色质细胞、多巴胺能或血清素能神经元或PC12细胞）放置在显微镜下观察，刺激细胞并记录一系列释放事件来完成的。该项目开发的技术改编自半导体行业，涉及开发CMOS微电子芯片，用于在芯片上记录可氧化传递分子（如去甲肾上腺素、多巴胺或血清素）的单量子释放事件。该技术将允许同时记录来自数百个细胞的单个囊泡释放事件，而不需要显微镜观察和操作，因此将提供一个高通量平台来表征分子和药理学操作。这项技术将加速旨在了解递质释放及其调节的分子机制的研究，以及通过调节递质释放来发挥作用的治疗方法的开发和测试。通过这种方式，这种新的分析平台将提供机会来测量定量释放事件作为神经生物学终点，并将为目标识别和药物发现提供管道。