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: 8190221
• 负责人: Manfred LINDAU
• 金额: $ 37.62万
• 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/8190221
• 关键词: Adrenal Glands; Affect; Amplifiers; Binding; Biological Assay; Botox; Brain; Catecholamines; Cell Communication; Cell Count; Cell Size; Cells; 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. PUBLIC HEALTH RELEVANCE: Release of neurotransmitters and hormones is regulated by many proteins, signaling molecules and various drugs. Two examples are the drug L-Dopa, used to treat Parkinson's disease, or the cosmetic BoTox treatment. 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 individual transmitter release events. This novel assay platform will 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）：该项目的目的是开发具有带有芯片放大器的可扩展的N X N电化学检测器阵列平台，以大量平行的量子发射机释放事件记录。该测定法将分析的神经生物学过程是胞外增生和发射器释放的过程，这是大脑功能及其他过程中的关键过程之一。要释放的分子存储在膜结合的细胞器中。刺激后，这些囊泡的含量通过融合孔将囊泡腔与细胞外空间联系起来。了解囊泡融合和发射器释放的机制具有广泛的医学意义。在帕金森氏病的治疗中，药物左旋多巴增加了多巴胺能神经元减少的多巴胺释放。另一方面，肉毒杆菌毒素治疗通过减少发射机释放来发挥其作用。除这些示例外，许多其他药物和许多分子操作以各种方式调节发射机释放。发射机释放的调节不仅是通过更改量化事件的数量或频率而发生的，而且还通过调节量化量和从单个囊泡释放的动力学进行调节。为了了解特定的操作影响发射机释放的机制，必须精确测量单个定量释放事件，分析其振幅和时间过程并得出特征参数。通常，通过将碳纤维微电极定位在微观观察下，刺激细胞并记录一系列释放事件，从而进行了这种测量。该项目开发的技术是根据半导体行业进行的，涉及开发CMOS微电源芯片，用于用于芯片录音的单次释放事件的芯片记录，例如氧化发射机分子，例如去甲肾上腺素，多巴胺或5-羟色胺。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，而无需微观观察和操纵，从而提供一个高通量平台来表征分子和药理操纵。该技术将加速旨在理解发射器释放的分子机制及其调节的分子机制，以及通过调节发射机释放来起作用的处理和测试。通过这种方式，这个新颖的测定平台将提供机会来衡量数量释放事件作为神经生物学终点，并为目标识别和药物发现提供管道。 公共卫生相关性：神经递质和激素的释放受许多蛋白质，信号分子和各种药物的调节。有两个例子是用于治疗帕金森氏病的药物L-DOPA或化妆品肉毒杆菌毒素治疗。该项目的目的是开发具有片上放大器的可扩展N X N电化学检测器阵列平台，以大量平行单个发射器释放事件。这个新颖的测定平台将提供机会，以作为神经生物学终点来衡量量化事件，并将为目标识别和药物发现提供管道。