Elucidating Synaptic Regulators via High-Throughput Morphology Characterization

通过高通量形态学表征阐明突触调节因子

• 批准号: 9213710
• 负责人: Adriana San Miguel
• 金额: $ 24.9万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9213710
• 关键词: Address; Affect; Age; Aging; Algorithms; Alzheimer' s Disease; Animals; Autistic Disorder; Automation; Behavioral; Biological; Biology; Caenorhabditis elegans; Chimeric Proteins; Cognitive; Computer Vision Systems; Cues; Data; Data Analyses; Defect; Descriptor; Deterioration; Development; Disease; Docking; Electron Microscopy; Engineering; Environment; Epigenetic Process; Exposure to; Fluorescence; Genes; Genetic; Genetic Screening; Goals; Health; Heterogeneity; Huntington Disease; Image; Image Analysis; Impaired cognition; Institution; Ion Channel; Label; Lead; Life; Light; Link; Locomotion; Longevity; Measures; Memory; Mental Depression; Mentors; Methods; Microfluidics; Microscopy; Modeling; Mood Disorders; Morphology; Motor; Movement; Mutation; Nematoda; Nerve Degeneration; Neurophysiology - biologic function; Neurosciences; Neurotransmitters; Parkinson Disease; Pathology; Pathway interactions; Pattern; Phase; Physiological; Play; Population; Preclinical Drug Evaluation; Presynaptic Terminals; Research; Role; Sampling; Schizophrenia; Signal Transduction; Site; Synapses; Synaptic Cleft; Synaptic Transmission; Synaptic Vesicles; Syncope; System; Techniques; Technology; Testing; Training; Work; age related; base; career development; cognitive function; excitatory neuron; in vivo; innovation; nervous system disorder; neurotransmission; neurotransmitter release; neurotransmitter uptake; normal aging; optogenetics; post-doctoral training; postsynaptic; presynaptic; receptor; relating to nervous system; response; synaptic function; technology development; tissue fixing; tool

DESCRIPTION (provided by applicant): Many neurological diseases occur in the absence of neurodegenerative pathology, such as neurotransmission disorders. Deficient neurotransmission is a hallmark of many neurological diseases, such as depression, schizophrenia and autism. Moreover, deterioration in synaptic function also appears during ageing, accompanied by a decline in cognitive and behavioral function. Synaptic strength and plasticity, important in cognitive functions such as memory, are affected by synaptic activity. However, the regulators of synaptic strength, either activity-dependent or independent, are far from understood. Here, I propose to explore how environmental cues and ageing can affect synaptic morphology in the nematode C. elegans, and how this correlates to synaptic function and activity. In C. elegans, which is an excellent model for neuroscience, synaptic sites can be observed with fluorescent fusion proteins. However, fluorescently labeled synaptic sites are small and faint, and obtaining large number of high-content data poses many experimental limitations. The main goal of this proposal is to elucidate regulators of synaptic function through multidimensional morphological profiling of synaptic sites. This work is composed of a mentored and an independent research phase. During the mentored phase, I will develop tools that allow high-throughput quantitative multidimensional profiling of synaptic morphology in large populations of animals. These tools are based on combining microfluidics, automation, and computer vision methods for image analysis, which enable streamlined quantitative morphological characterization of synaptic sites. In addition to this, tools to determine synaptic function in the motor circuit through the quantification of locomotive activity and controlled stimulation of excitatory neurons will be developed to correlate synaptic morphology to function. The mentored phase will also include extensive training in various aspects: technology development, biology and neuroscience, and career development. The scientific environment at the institution of the mentored phase is highly collaborative and provides excellent support in terms of facilities and intellectual opportunities. During the independent research phase, I plan to apply the developed tools during the mentored phase to uncover how environmental cues can affect synaptic function through activity dependent or independent mechanisms. I will also utilize these tools to study synaptic decline during aging, and how exposure to environmental regulators of synaptic function during development can alter synaptic decline during aging. In this way, we will be able to address a biological question unapproachable with conventional methods. This approach is innovative not only because the technology developed will greatly increase the throughput and quality of characterization, but also because it proposes studying the links between form and function in synaptic sites. This project is significant because it will enable streamlined morphological studies in neuroscience, which should lead to uncovering pathways, genes and potential therapies for synaptic function deficiencies due to disease or age-related decline.

描述（由申请人提供）：许多神经疾病在没有神经退行性病理（例如神经传递疾病）的情况下发生。缺乏神经传递是许多神经系统疾病的标志，例如抑郁症，精神分裂症和自闭症。此外，在衰老期间，突触功能的恶化也会出现，伴随着认知和行为功能的下降。突触强度和可塑性在认知功能（例如记忆）中很重要，受到突触活动的影响。但是，依赖活性或独立的突触强度的调节剂远非理解。在这里，我建议探索环境线索和衰老如何影响线虫秀丽隐杆线虫中的突触形态，以及该如何与突触功能和活动相关。在秀丽隐杆线虫（C. C. c. ofe）中，这是神经科学的极好模型，可以使用荧光融合蛋白观察突触部位。但是，荧光标记的突触部位小而微弱，获得大量的高含量数据构成了许多实验局限性。该提案的主要目标是通过 突触部位的多维形态分析。这项工作由一个指导和独立的研究阶段组成。在指导阶段，我将开发工具，以允许大量动物种群中突触形态的高通量定量多维分析。这些工具基于组合图像分析的微流体，自动化和计算机视觉方法，该方法可以简化突触部位的定量形态特征。除此之外，还将开发通过量化机车活性和受控刺激的兴奋性神经元的工具来确定运动电路中的突触功能，以将突触形态与功能相关联。指导阶段还将包括各个方面的广泛培训：技术开发，生物学和神经科学以及职业发展。指导阶段机构的科学环境是高度协作的，并在设施和智力机会方面提供了出色的支持。在独立研究阶段，我计划在指导阶段应用开发的工具，以发现环境线索如何通过依赖于活动或独立机制影响突触功能。我还将利用这些工具来研究衰老期间的突触下降，以及在发育过程中对突触功能的环境调节剂的暴露如何改变衰老期间的突触下降。这样，我们将能够通过传统方法解决一个无法接近的生物学问题。这种方法具有创新性，这不仅是因为开发的技术会大大提高表征的吞吐量和质量，还因为它提出了研究突触部位中形式和功能之间的联系。该项目具有重要意义，因为它将在神经科学中进行简化的形态学研究，这应该导致由于疾病或与年龄相关的下降而导致突触功能缺陷的途径，基因和潜在疗法。