Microfluidic Platform for Probing Ceramide Channels

用于探测神经酰胺通道的微流控平台

• 批准号: 7641919
• 负责人: Don L DeVoe
• 金额: $ 21.33万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7641919
• 关键词: Address; Affect; Apoptosis; Area; Basic Science; Behavior; Binding; Binding Sites; Biological; Biophysics; Buffers; Caliber; Cell membrane; Cell physiology; Cells; Ceramides; Commit; Cytosol; Data; Data Collection; Decision Making; Defect; Detection; Development; Devices; Diffusion; Dimensions; Disease; Electric Capacitance; Electrodes; Electronics; Electrophysiology (science); Elements; Entropy; Experimental Designs; Film; Gel; Heating; Human Resources; In Situ; Individual; Investigation; Ion Channel; Ions; Kinetics; Knowledge; Lead; Lipids; Location; Manuals; Measurement; Measures; Mechanics; Membrane; Microfluidics; Mitochondria; Mitochondrial Proteins; Molecular; Molecular Probes; Monitor; Nature; Noise; Operative Surgical Procedures; Pattern; Performance; Perfusion; Phospholipids; Polymers; Process; Property; Protein Family; Proteins; Pulse Pressure; Reagent; Regulation; Reporting; Research; Research Personnel; Resistance; Role; Side; Silicon; Site; Sodium Chloride; Solutions; Sphingolipids; Structure; System; Technology; Teflon; Temperature; Testing; Thermodynamics; Time; Training; Transport Process; Variant; analytical tool; aqueous; base; cost; design; enthalpy; flexibility; inhibitor/antagonist; instrumentation; macromolecule; novel; pressure; prevent; public health relevance; research study; response; single molecule; small molecule; temperature jump; time use; tool; voltage; voltage clamp

DESCRIPTION (provided by applicant): A microfluidic platform is proposed that will aid in investigating processes involved in lipid channel formation and dissolution, leading to a deeper understanding of molecular transport across cell membranes, and ultimately contribute to our knowledge of biological channels and their relationships to disease processes. The project will leverage recent results from our team, including the first demonstration of single-molecule detection within a microfluidic ion channel chip using a single membrane-bound biological ion channel as a conductometric sensing element, with real-time control of analyte concentration and buffer conditions on either side of the channel. In this proposal, the proof-of-concept microfluidic platform will be extended to demonstrate in situ phospholipid membrane formation, integrated supporting gels and microporous films for enhanced membrane stability, automated multiplexed lipid channel measurements, and thin film thermal control at the membranes to enable the observation of the relationships between enthalpy and membrane formation, channel formation and dynamics, and analyte/channel interactions. A range of experiments that are not feasible or extremely time-consuming with existing technology will be conducted, with a focus on investigating channels formed by ceramide, a sphingolipid implicated in apoptosis. The structure and function of ceramide channels will be probed using the microfluidic system, with controlling factors manipulated to investigate their impacts on the regulation of channel size and stability. This effort will result in a novel and unique electrophysiology platform which may be applied to a wide range of fundamental and applied investigations of biological channels, together with experimental results that will expand our understanding of an important lipid channel system involved in programmed cell death. PUBLIC HEALTH RELEVANCE: A unique experimental platform is proposed that will elucidate the processes involved inbiological membrane channel formation and dissolution, leading to a deeper understanding of molecular transport across cell membranes, and ultimately contribute to our knowledge of biological channels and their relationships to disease processes.

描述（由申请人提供）：提出了一种微流体平台，该平台将有助于研究脂质通道形成和溶解所涉及的过程，从而更深入地了解跨细胞膜的分子运输，并最终有助于我们了解生物通道及其与疾病过程的关系。该项目将利用我们团队的最新成果，包括首次演示微流控离子通道芯片内的单分子检测，使用单个膜结合生物离子通道作为电导传感元件，实时控制通道两侧的分析物浓度和缓冲条件。在该提案中，概念验证微流控平台将扩展到演示原位磷脂膜形成、用于增强膜稳定性的集成支撑凝胶和微孔膜、自动多重脂质通道测量以及膜上的薄膜热控制，以能够观察焓与膜形成、通道形成与动力学以及分析物/通道之间的关系 互动。将进行一系列现有技术不可行或极其耗时的实验，重点是研究神经酰胺（一种与细胞凋亡有关的鞘脂）形成的通道。将使用微流体系统探测神经酰胺通道的结构和功能，并操纵控制因素来研究它们对通道尺寸和稳定性调节的影响。这项工作将产生一个新颖而独特的电生理学平台，该平台可应用于生物通道的广泛基础和应用研究，以及实验结果，将扩大我们对参与程序性细胞死亡的重要脂质通道系统的理解。公共健康相关性：提出了一个独特的实验平台，该平台将阐明生物膜通道形成和溶解所涉及的过程，从而更深入地了解跨细胞膜的分子运输，并最终有助于我们了解生物通道及其与疾病过程的关系。