ULTRASTRUCTURAL BASIS FOR SYNAPTIC VESICLE RECYCLING IN CALYX OF HELD

持有花萼突触小泡回收的超微结构基础

• 批准号: 8361902
• 负责人: GEORGE A SPIROU
• 金额: $ 3.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361902
• 关键词: Area; Auditory; Brain Stem; Cell Nucleus; Cells; Characteristics; Communication; Complex; Elements; Filament; Funding; Goals; Grant; Growth Cones; Image; Image Analysis; Laboratories; Link; Medial; Mitochondria; Mono-S; Mus; National Center for Research Resources; Nerve; Organelles; Postsynaptic Membrane; Principal Investigator; Recycling; Research; Research Infrastructure; Resolution; Resources; Shapes; Sound Localization; Source; Staging; Structure; Synapses; Synaptic Vesicles; Time; Transmission Electron Microscopy; United States National Institutes of Health; Work; base; cost; digital; electron tomography; fluorescence imaging; metaplastic cell transformation; nerve supply; neuronal cell body; neurotransmission; postnatal; postsynaptic; presynaptic; reconstruction; research study; trapezoid body

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. ULTRASTRUCTURAL BASIS FOR SYNAPTIC VESICLE RECYCLING IN THE CALYX OF HELD The calyx of Held and other large nerve terminals of the auditory brainstem are key elements of sound localization circuitry. Our goal is to reveal structural transformations and cellular communication that characterize contact of the calycigenic growth cone with its target and the early stages of synapse assembly and stabilization at large nerve terminals in the auditory brainstem. Our central hypothesis is that competition among calycigenic inputs precedes expansion of the terminal over the cell body to form a calyx. This hypothesis is based on work from our laboratory that reveals rapid formation of the calyx in mice between postnatal days (P)2 and P4. The calyx contains hundreds to greater than two thousand active zones, depending upon the species, many of which are located nearby specialized organelle complexes termed mitochondrion-associated adherens complexes (MACs). MAC structure had been described previously using standard transmission electron microscopy (sTEM), which revealed filaments tethering the mitochondrion to a punctum adherens that links the pre- and postsynaptic membranes. Confocal fluorescence imaging and electron tomography are being employed to study medial nucleus of the trapezoid body (MNTB) cell innervation during P0P4. This time period precedes and overlaps the formation of immature, cup-shaped calyces that envelop the MNTB cell body. These experiments will pinpoint time periods during which mono-innervation is established between pre- and postsynaptic partners and highlight structural and functional differences that may predict winning and losing inputs. In addition, we will describe structural features of mature calyces that support high-rate neurotransmission that is characteristic of this terminal. Because of the size of the calyx-MNTB contact and the desired high resolution of the reconstructions, serial volumetric imaging of domains of cells will be required. This project will require high-resolution, wide-field, large-area digital recording of images for electron tomography.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 在静置的花萼中进行突触囊泡回收的超微结构基础 听觉脑干的固定和其他大神经末端的花萼是声音定位电路的关键要素。我们的目标是揭示结构转化和细胞通信，这些结构转化和细胞通信表征了钙化生长锥与其靶标的接触以及突触组装的早期阶段和在听觉脑干大神经末端的早期阶段。我们的中心假设是，钙化输入之间的竞争是末端在细胞体上膨胀以形成花萼的竞争。该假设是基于我们实验室的工作，该工作揭示了产后（P）2和P4之间小鼠中花萼的快速形成。乳萼含有数百到大于2000多个活性区，具体取决于该物种，其中许多位于附近的专用细胞体配合物，称为线粒体相关的粘附剂络合物（MAC）。先前使用标准透射电子显微镜（STEM）描述了MAC结构，后者揭示了将线粒体绑定到连接前后突触后膜的点子粘附物上的细丝。 在P0 P4期间，正在使用共聚焦荧光成像和电子断层扫描来研究梯形体（MNTB）细胞神经支配的内侧核。这段时间之前并重叠了包围MNTB细胞体的未成熟，杯形的钙的形成。这些实验将确定在伴侣和突触后伴侣之间建立单一损害的时间段，并突出显示可能预测获胜和输入输入的结构和功能差异。此外，我们将描述支持该终端特征的高速神经传递的成熟钙的结构特征。由于花萼-MNTB接触的大小和重建所需的高分辨率，因此需要对细胞域的串行体积成像。该项目将需要电子断层扫描图像的高分辨率，广阔的大区域数字记录。