3D FINE STRUCTURE OF THE NEUROMUSCULAR JUNCTION

描述（来自申请人的摘要）为了检验假设， 在神经肌肉接头处构成突触器的蛋白质具有 一个具体而有序的安排，PI和他的同事已经开始 开发使用电磁层析成像的方法， 组织切片的重建。 从这种重建中获得的数据 提供了丰富的新的定量信息来源， 突触器的组成和组织小于4nm 三轴分辨率。 免疫金标记的位置 突触蛋白也可以被精确地确定。 的具体目标 该提案中概述的实验是： 1. 评估不同的组织制备方法、数据收集方法， 重建和代表性的研究精细结构的 神经肌肉突触器的三维模型。 2. 表征装置的3D精细结构。 3. 为了在3D中映射特定突触的免疫金标记的位置， 蛋白质与器官的特定结构成分。 神经肌肉的分子结构的详细知识 连接对于理解突触相关机制至关重要 传输和维护。 它也是必要的，以确定如何 神经肌肉接头在个体发育过程中形成，并在创伤后再生 以及用于表征发病机制。 EM层析成像提供了 获得这种信息的机会在一个显着较高的水平， 分辨率比目前可用的任何其他方法或 眼前的地平线。 此外，由于它也可以用于研究3D 身体任何组织的精细结构，对其进行系统研究的方法 在神经肌肉接头处的应用，如这里所提出的，应该提供 这些见解将加速其他人进入这一新兴领域， 研究方法

DESCRIPTION (from the applicant's abstract) To test the hypothesis that proteins composing the synaptic apparatus at the neuromuscular junction have a specific and orderly arrangements, the PI and his colleagues have begun developing ways of using EM tomography for making accurate 3D reconstructions of tissue sections. Data obtained from such reconstructions furnish a rich new source of quantitative information about the structural composition and organization of the synaptic apparatus to less than 4nm resolution in all three axes. The position of immunogold labels for synaptic proteins can also be accurately determined. The specific aims of the experiments outlines in this proposal are: 1. To assess different methods of tissue preparation, data collection, reconstruction and representation for examining fine structure of the neuromuscular synaptic apparatus in 3D. 2. To characterize the 3D fine structure of the apparatus. 3. To map in 3D the position of immunogold labels for specific synaptic proteins to specific structural components of the apparatus. A detailed knowledge of the molecular architecture of the neuromuscular junction is essential for understanding the mechanisms involved in synaptic transmission and maintenance. It is also necessary for determining how the neuromuscular junction forms during ontogenesis and regenerates after trauma and for characterizing mechanisms of pathogenesis. EM tomography offers the opportunity of obtaining such information at a significantly higher level of resolution than that provided by any other method currently available or on the immediate horizon. Moreover, since it can also be used to study the 3D fine structure of any tissue in the body, a systematic approach to its application at the neuromuscular junction, as proposed here, should provide insights that will expedite the entry of others into this nascent field of research methodology.