THREE DIMENSIONAL CHARACTERIZATION OF PNC IN MAMMALIAN CELL NUCLEI

这项工作的重点是结构功能的分析 脑内已识别的突触前神经元和突触后神经元之间的关系 映射的感官系统。我们计划用IVEM来做连续剧 初级传入和感觉中间神经元的重建 确定突触的位置、数量和类型 这些细胞。我们还将使用3-D重建，使用序列 体层摄影术估计大脑皮质的体积和功能尺寸 在两种细胞类型的特定解剖区域中的突触。 中间神经元特定区域大小的测量 将被用来估算隔室的膜表面积 模特儿研究。因为这些细胞很大，而且 重要的是对细胞的许多不同区域进行采样，常规 薄片显微镜将使这项任务非常耗时。 而且很难。在IVEM中使用更厚的部分的能力将 使我们能够在必要的情况下快速获得此信息 精确度。IVEM的研究将补充正在进行的分析 该系统处于光学显微镜级别。我们还计划使用 用IVEM研究新突触的形成和生长 在发育过程中和在特定条件下饲养的动物中的传入 刺激环境。这个项目的工作曾短暂中断 当雅各布博士和米勒博士搬到蒙大拿州建立新的 他们是计算生物学中心的联合主任。 然而，我们已经开始分析荧光标记的 在共聚焦显微镜上的标本，并希望扩大这种分析 到电子显微镜的水平在不久的将来。此外，我们 正在研究用于执行自动骨架分析的软件。

This work focuses on the analysis of structure function relationships between identified pre and postsynaptic neurons in a mapped sensory system. We plan to use the IVEM to do serial reconstructions of primary afferents and sensory interneurons to determine the location, number and type of synapses on and between these cells. We will also use 3-D reconstructions using serial tomography to estimate the volume and functional dimensions of synapses in specific anatomical regions of both cell types. Measurements of the dimensions of specific regions of interneurons will be used to estimate membrane surface area for compartmental modeling studies. Since these cells are quite large and it is important to sample many different areas of the cell, conventional thin section microscopy would make this task extremely time consuming and difficult. The ability to use thicker sections in the IVEM will enable us to obtain this information rapidly and with the necessary accuracy. The IVEM studies will compliment an ongoing analysis of this system at the light microscope level. We also plan to use the IVEM to study the formation and growth of new synapses on primary afferents during development and in animals reared in specific stimulus environments. Work on this project was interrupted briefly while Drs. Jacob and Miller moved to Montana to establish the new Center for Computational Biology, of which they are co-directors. Nevertheless, we have begun analysis of fluorescently labeled specimens on the confocal microscope and hope to extend this analysis to the electron microscopic level in the near future. In addition, we are investigating software for performing automated skeletonization.