Imaging synapse formation using novel microfluidic platforms

使用新型微流体平台对突触形成进行成像

• 批准号: 8306755
• 负责人: Deyu Li
• 金额: $ 23.37万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8306755
• 关键词: Actins; Address; Axon; Brain; Cell Communication; Cell Culture Techniques; Cells; Coculture Techniques; Communication; Complex; Cytoskeleton; DNA Sequence Rearrangement; Dendrites; Dendritic Spines; Development; Device Designs; Devices; Environment; Excitatory Synapse; Family; Fluorescence Resonance Energy Transfer; Guanosine Triphosphate Phosphohydrolases; Image; Indium; Individual; Intercellular Junctions; Lead; Life; Mediating; Microfluidic Microchips; Microfluidics; Microscopy; Modification; Molecular; Neuraxis; Neuroglia; Neurons; Peripheral Nervous System; Play; Presynaptic Terminals; Process; Proteins; Regulation; Resolution; Role; Signal Transduction; Staging; Structure; Synapses; Techniques; Technology; Testing; Time; Vertebral column; base; cognitive function; design; image processing; in vivo; innovation; interest; nervous system disorder; neuromuscular; novel; photoactivation; postsynaptic; research study; rho; rho GTP-Binding Proteins; selective expression; spatiotemporal; synaptic function; synaptogenesis

DESCRIPTION (provided by applicant): Synapses are highly specialized cell-cell junctions that mediate communication between neurons. These structures are composed of pre- and post-synaptic terminals and are the basis for the complex circuitry found in the brain. Most postsynaptic terminals of excitatory synapses take the form of dendritic spines, which are actin-rich protrusions that emanate from the dendrite shaft. Not surprisingly, the formation and plasticity of dendritic spines and synapses play a central role in cognitive function and abnormalities in these structures are associated with a number of neurological disorders. Despite the importance of spines and synapses in the central nervous system, the molecular mechanisms that regulate the formation of these structures are not well understood. A limitation toward identifying key molecules that regulate spine and synapse formation has been the great difficulty in observing synapses as they form. We are developing novel microfluidic devices that will allow us to dynamically observe forming synapses (Specific Aim I). Several innovations in the design of these devices will significantly enhance our ability to image the early steps of synapse formation with high spatial and temporal resolution. In Specific Aim II, we will apply this technology to examining the spatiotemporal dynamics of actin during synaptic assembly. In addition, we will test our hypothesis that the activity of Rho family GTPases, which are key regulators of actin, is critical in the initial assembly and maturation of synapses. For these experiments, we will use cutting-edge microscopy technologies, including FRAP, photoactivation, and FRET to examine actin dynamics and regulation during synapse formation. The development of these microfluidic platforms will be of great interest and benefit to neurobiologist by providing a platform for identifying the key molecular signals that regulate the assembly of synapses.

描述（由申请人提供）：突触是高度专业化的细胞间连接，介导神经元之间的通信。 这些结构由突触前和突触后末梢组成，是大脑中复杂电路的基础。 大多数兴奋性突触的突触后末端呈树突棘的形式，树突棘是从树突轴发出的富含肌动蛋白的突起。 毫不奇怪，树突棘和突触的形成和可塑性在认知功能中发挥着核心作用，这些结构的异常与许多神经系统疾病有关。 尽管棘和突触在中枢神经系统中很重要，但调节这些结构形成的分子机制尚不清楚。 识别调节脊柱和突触形成的关键分子的限制是观察突触形成的巨大困难。 我们正在开发新型微流体装置，使我们能够动态观察突触的形成（具体目标 I）。 这些设备设计中的几项创新将显着增强我们以高空间和时间分辨率对突触形成的早期步骤进行成像的能力。 在Specific Aim II中，我们将应用这项技术来检查突触组装过程中肌动蛋白的时空动态。 此外，我们将检验我们的假设，即 Rho 家族 GTP 酶（肌动蛋白的关键调节因子）的活性对于突触的初始组装和成熟至关重要。 在这些实验中，我们将使用尖端显微镜技术，包括 FRAP、光激活和 FRET 来检查突触形成过程中的肌动蛋白动力学和调节。 这些微流体平台的开发将引起神经生物学家的极大兴趣并受益，因为它们提供了一个识别调节突触组装的关键分子信号的平台。