MACROMOLECULAR ARCHITECTURE OF THE SYNAPSE

这个新项目分析了分子结构。 对哺乳动物大脑中突触的研究，最初的重点是 突触后密度(PSD)。它的许多分子成分 PSD是已知的，但PSD是如此巨大(大约10到 第九大)认为必须开发新的方法来理解 它的众多分子成分是如何组织起来的。一个 前景看好的新方法开始于将PSD连接到玻璃和 然后冷冻并复制它们进行电子显微镜检查。 释放之前的操作允许识别和本地化 特定的部件。这些问题包括：私营部门司的定位 在玻璃上预先处理含有一种抗体的玻璃 表面成分；部分消化PSD上的 含洗涤剂的玻璃；以及在上述任何一项之后贴上标签 用胶体金标记的抗体。到目前为止的工作很清楚 区分PSD的裂侧，其中NR-1和PSD-95 定位于CaMKII所在的细胞质一侧。 对裂隙侧的消化揭示了形成的晶格状结构 PSD的核心；这个格子为PSD-95贴上了沉重的标签。这个 幽灵蛋白和肌动蛋白在PSD中的分布目前正在 已调查，CaMKII的分布是 另一个项目。现在很明显，地球表面的空间分布 PSD组件是PSD功能的一个重要方面。

This new project analyzes the molecular architecture of synapses in the mammalian brain with an initial focus on the postsynaptic density (PSD). Many molecular components of the PSD are known, but the PSD is so enormous (approximately 10 to the ninth Da) that new methods had to be developed to understand how its numerous molecular components are organized. A promising new approach begins by attaching PSDs to glass and then freezing and replicating them for electron microscopy. Manipulations prior to freeing allow identification and localization of particular components. These include: orientation of the PSDs on the glass by prior treatment of the glass with antibodies to one of their surface components; partial digestion of the PSDs on the glass with detergents; and labelling, after either of the above with antibodies marked by colloidal gold. Work to date clearly distinguishes the cleft side of the PSD, where NR-1 and PSD-95 are localized, from the cytoplasmic side where CaMKII is localized. Digestion of the cleft side reveals a lattice-like structure that forms the core of the PSD; this lattice labels heavily for PSD-95. The distributions of spectrin and actin with the PSD are currently being investigated, and the distribution of CaMKII is the subject of another project. It is now apparent that the spatial distributions of components of the PSD are an important aspect of PSD function.