High Resolution Analysis of miR125b in Dendrites via Microfluidic Devices

通过微流体装置对树突中的 miR125b 进行高分辨率分析

• 批准号: 8571230
• 负责人: Martha U Gillette
• 金额: $ 23.79万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8571230
• 关键词: Address; Affective; Aging; Alzheimer' s Disease; Autistic Disorder; Back; Behavior; Binding; Biological Assay; Biological Process; Brain; Cell physiology; Cellular biology; Chronic stress; Complex; Cues; Defect; Dendrites; Development; Developmental Therapeutics Program; Disease; Disease Progression; Engineering; Environment; FMRP; Filopodia; Fragile X Syndrome; Functional RNA; Functional disorder; Future; Glutamates; Goals; Growth; Heterogeneity; Hippocampus (Brain); Human; Image; Image Analysis; Individual; Knowledge; Maintenance; Maps; Measures; Mediating; Mental Retardation; Methods; MicroRNAs; Microfabrication; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Molecular Profiling; Morphology; Nervous System Physiology; Nervous system structure; Neurites; Neurons; Neurosciences; Outcome; Play; Population; Process; Property; Protein Biosynthesis; Protocols documentation; Public Health; Regulation; Research; Resolution; Role; Schizophrenia; Science; Signal Transduction; Stimulus; Structure; System; Techniques; Technology; Therapeutic; Variant; Vertebral column; Work; age related; density; design; developmental disease; experience; innovation; insight; nanolitre; neuron development; new technology; novel; novel diagnostics; protein expression; public health relevance; research study; response; synaptogenesis; tool

DESCRIPTION (provided by applicant): Spatially defined sub-cellular heterogeneity determines neuron function. Thus, it is not surprising that disease origins can be traced back to the aberrant behavior(s) of dendritic filopodia that wire the brain. Interactions of the myriad filopodia extended by dendrites of individual neurons in their spatial contexts generate the remarkable range of functionalities of the human brain. Even adjacent filopodia encounter distinct local micro-environments and develop individual functionalities. Only by overcoming ensemble averaging of populations and measuring molecular signatures with single- filopodium resolution can we understand the interplay of the diverse intrinsic and extrinsic regulators, and explain the spectrum of neurological functions encompassing healthy and disease states. In particular, there is an unmet need for ways of probing of local regulators of filopodia during the emergence and sculpting of the dendritic arbor. This innovation proposal addresses this need by integrating our expertise in designing and fabricating nanoliter microfluidic environments for ultra-low density neuronal cultures with our expertise in the cell biology of neurons. We propose to use microfluidic device (¿FD) environments and high resolution image analysis to probe changes in localization, activity, and function of specific microRNAs (miRNAs) in developing hippocampal dendrites. miRNAs are short, non-coding RNAs that act as regulators of local protein synthesis, especially during dendrogenesis and local wiring of the nervous system. Our objective is to control the structure and function of individual dendrites within micro-channels of fabricated ¿FDs to isolate individual dendrites. We will use this system to map and influence miR125b functioning in filopodia during their development and in response to glutamatergic stimulation. This novel set of studies will address the need for understanding with high resolution the localization, activation, and function of miR125b during wiring of the hippocampus. This approach will provide new insights on this putative regulator, new tools for studying properties of miRNA control of dendrogenesis in single neurons, and contribute to effective strategies for restoring defects in models of affective dysfunctions, chronic stress, Alzheimer's disease, and autism.

描述（适用提供）：空间定义的亚细胞异质性决定了神经元功能。这是毫不奇怪的是，疾病起源可以追溯到大脑丝线的树突状丝状疾病的异常行为。各个神经元在其空间环境中的树突扩展的无数丝状疾病的相互作用产生了人脑的显着功能范围。即使相邻的丝状疾病也会遇到不同的局部微环境并发展个体功能。只有通过克服人群的平均合奏并用单丝苯基分辨率测量分子特征，我们才能理解潜水员的固有和外在调节剂的相互作用，并解释神经功能的频谱，包括健康和疾病状态。特别是，在出现和雕刻树突状植物园期间，未满足探测菲洛修的局部调节剂的需求。这项创新提案通过整合我们在设计和制造纳米素的微流体环境方面的专业知识来满足这一需求，以实现超低密度神经元培养物与我们在神经元细胞生物学方面的专业知识。我们建议使用微流体设备（FD）环境和高分辨率图像分析来探测特定microRNA（miRNA）在开发海马树突中的定位，活性和功能的变化。 miRNA是简短的非编码RNA，可作为局部蛋白质合成的调节剂，尤其是在神经系统的树突生成和局部接线期间。我们的目标是控制单个树突的结构和功能 制造的»fds以隔离单个树突。我们将使用该系统来绘制和影响MiR125b在发育过程中在其发育过程中的功能并响应谷氨酸能刺激。这项新的研究集将解决高分辨率在海马接线期间MiR125b的定位，激活和功能的必要性。这种方法将为这种推定的调节剂提供新的见解，用于研究单个神经元中树突生成的特性的新工具，并为恢复情感功能障碍，慢性压力，阿尔茨海默氏病和自闭症模型的缺陷有效策略有效。