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（由申请人提供）：空间定义的亚细胞异质性决定了神经元功能，因此，疾病起源可以追溯到连接大脑的无数丝状伪足的异常行为也就不足为奇了。单个神经元的树突在其空间环境中延伸，产生了人类大脑的一系列非凡的功能，甚至相邻的丝状伪足也会遇到不同的局部微环境并发育出个体。只有通过克服群体的整体平均并以单丝足分辨率测量分子特征，我们才能了解不同的内在和外在调节因子的相互作用，并解释涵盖健康和疾病状态的神经功能谱。在树突乔木的出现和雕刻过程中，对丝状伪足局部调节剂的探测方法的需求未得到满足。这项创新提案通过整合我们在设计和制造纳升微流体环境方面的专业知识来满足这一需求。凭借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装置（¿ FD）环境和高分辨率图像分析，以探测海马树突中特定 microRNA (miRNA) 的定位、活性和功能的变化 miRNA 是短的、非编码的发育中的 RNA，充当局部蛋白质合成的调节剂，尤其是在树突形成和发育过程中。我们的目标是控制神经系统微通道内单个树突的结构和功能。 捏造 ¿我们将使用该系统来绘制和影响 miR125b 在丝状伪足发育过程中的功能以及响应谷氨酸刺激的需求。这种方法将为研究 miRNA 对单个神经元树突状细胞生成的控制特性提供新的见解。恢复情感功能障碍、慢性压力、阿尔茨海默病和自闭症模型缺陷的有效策略。