Simultaneous kinetic analyses of neuronal connectivities

神经元连接的同步动力学分析

• 批准号: 9048554
• 负责人: Ernest Fitch Guignon
• 金额: $ 15.03万
• 依托单位: CIENCIA, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-15 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9048554
• 关键词: Accounting; Affect; Antibodies; Antigens; Area; Autistic Disorder; Automation; Axon; Biological Assay; Biosensor; Brain; Brain region; Cells; Communication; Computer software; Corpus striatum structure; Coupled; Development; Devices; Disease; Extracellular Matrix; Extracellular Matrix Proteins; Fluorescence; Fluorochrome; Geometry; Goals; Gold; Immunofluorescence Immunologic; Infection; Kinetics; Label; Laboratories; Location; Measures; Mental Health; Microfluidics; Microglia; Microscopic; Modeling; Molecular; Monitor; Morphology; Mus; Neurons; Neurotransmitters; Parkinson Disease; Phenotype; Presynaptic Terminals; Proteins; Sampling; Signal Transduction; Site; Spottings; Staging; Substantia nigra structure; Surface Plasmon Resonance; Techniques; Technology; Therapeutic; Therapeutic Intervention; Time; Tissues; Work; antibody conjugate; axonal guidance; base; brain tissue; density; design; experience; fluid flow; follow-up; genetic analysis; instrument; instrumentation; neuroinflammation; neuronal circuitry; neuropsychiatric disorder; neurotoxicity; physical conditioning; postsynaptic; presynaptic; prototype; public health relevance; response; surface plasmon coupled emission; toxicant

 DESCRIPTION (provided by applicant): Aberrant neuronal connectivity is associated with a number of neuropathological and neuropsychiatric diseases/disorders. Inefficient developmental connectivity, misdirected connectivity, or disrupted and degenerated connectivity can account for losses of normal neuronal communication and subsequent losses of mental or physical health. Altered expression of axonal guidance proteins, neuroinflammation, neuronal toxicity and glial disturbances may be involved in inadequate neuronal connectivity. The goal of the work described in this application is to develop an instrument platform that can both enumerate development or loss of axons and the kinetics of these changes in response to molecular and cellular modifiers. The proposed grating coupled surface plasmon resonance (GCSPR) and grating coupled surface plasmon coupled emission (GCSPCE) instrumentation with a microflow biosensor chip will be developed and validated for quantification of developmental axonal outgrowths among punches of brain regional sections as well as loss of connections previously established. Up to five 1-3mm punches can be placed on the 1 cm2 biosensor gold chip precoated with extracellular matrix proteins and specific molecules (guidance proteins) or cells (microglia) spotted at specific locations. Lack or loss of axonal connections will be kinetically monitored by SPR and presynaptic and postsynaptic antigens can be assayed by SPCE with fluorochrome conjugated antibodies to neuronal antigens. The geometrical placement of specific brain regions will be identified so that preferential regional interactions can be quantified. The planned device to assess neuronal circuitry is built upon extensive experience with SPR microarrays that can characterize presence of single cells and their released products. The work described will produce instrumentation capable of providing a more complete and coherent picture of differential neuronal connectivities to provide a more comprehensive view of neuronal communications, which will reveal new opportunities for therapeutic interventions.

 描述（由申请人提供）：异常神经元连接与许多神经病理学和神经精神疾病/障碍相关。低效率的发育连接，错误的连接，或中断和退化的连接可以解释正常神经元通信的损失和随后的精神或身体健康的损失。轴突导向蛋白的表达改变、神经炎症、神经元毒性和神经胶质紊乱可能与神经元连接不足有关。本申请中描述的工作的目标是开发一种仪器平台，其可以计数轴突的发育或损失以及响应于分子和细胞修饰剂的这些变化的动力学。提出的光栅耦合表面等离子体共振（GCSPR）和光栅耦合表面等离子体耦合发射（GCSPCE）仪器与微流生物传感器芯片将开发和验证的定量发育轴突生长之间的冲床的大脑区域部分，以及以前建立的连接的损失。可以将多达五个1- 3 mm的穿孔物放置在预先涂覆有细胞外基质蛋白和特定分子（指导蛋白）或在特定位置点样的细胞（小胶质细胞）的1cm 2生物传感器金芯片上。轴突连接的缺乏或丧失将通过SPR进行动力学监测，突触前和突触后抗原可以通过SPCE用荧光染料缀合的神经元抗原抗体进行测定。将确定特定脑区的几何位置，以便可以量化优先区域相互作用。评估神经元回路的计划装置是建立在SPR微阵列的广泛经验之上的，SPR微阵列可以表征单细胞及其释放产物的存在。所描述的工作将产生能够提供更完整和连贯的图片差分神经元连接的仪器，以提供更全面的神经元通信，这将揭示新的治疗干预的机会。