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神经元功能。因此，疾病起源可以追溯到连接大脑的树突状丝状伪足的异常行为并不奇怪。由单个神经元树突延伸的无数丝状伪足在其空间环境中的相互作用产生了人类大脑的显着功能范围。甚至相邻的丝状伪足也会遇到不同的局部微环境，并发展出各自的功能。只有通过克服群体的系综平均和测量具有单丝状伪足分辨率的分子特征，我们才能理解多种内在和外在调节剂的相互作用，并解释包括健康和疾病状态的神经功能谱。特别是，在树枝状乔木的出现和雕刻期间，对丝状伪足的局部调节剂的探测方法存在未满足的需求。这项创新提案通过将我们在设计和制造用于超低密度神经元培养的纳升微流体环境方面的专业知识与我们在神经元细胞生物学方面的专业知识相结合来满足这一需求。我们建议使用微流控装置（FD）环境和高分辨率图像分析来探测特定microRNAs（miRNAs）在发育中海马树突的定位、活性和功能的变化。miRNA是短的非编码RNA，其充当局部蛋白质合成的调节剂，特别是在神经系统的树突发育和局部布线期间。我们的目标是控制微通道内单个树突的结构和功能， 制造FD以分离单个树突。我们将使用这个系统来映射和影响miR125 b在丝状伪足发育过程中的功能，并对神经元刺激做出反应。这组新的研究将解决高分辨率理解海马布线过程中miR125 b的定位、激活和功能的需求。这种方法将为这种假定的调节剂提供新的见解，为研究单个神经元中树突发育的miRNA控制特性提供新的工具，并有助于恢复情感功能障碍，慢性应激，阿尔茨海默病和自闭症模型中缺陷的有效策略。