MACROMOLECULAR ARCHITECTURE OF THE SYNAPSE

这个新项目分析了 的突触在哺乳动物的大脑与最初的重点放在 突触后密度（PSD）。许多分子组成的 PSD是已知的，但是PSD是如此巨大（大约10到100个）。 第九天）必须开发新的方法来理解 其众多的分子组成是如何组织的。一 一种很有前途的新方法是将PSD贴在玻璃上， 然后冷冻并复制它们用于电子显微镜检查。 释放前的操作允许识别和定位 特定的组件。这些问题包括： 在玻璃上预先用一种抗体处理玻璃 其表面成分;部分消化的PSD上的 玻璃与洗涤剂;和标签后，上述任何一个与 胶体金标记的抗体。工作日期明确 区分PSD的裂侧，其中NR-1和PSD-95 从CaMKII定位的细胞质侧定位。 对裂面的消化揭示了一种格子状结构， PSD的核心;这个格子标记为PSD-95。的 血影蛋白和肌动蛋白与PSD的分布目前正在 研究，CaMKII的分布是研究的主题。 另一个项目。现在很明显， PSD的组件是PSD功能的重要方面。

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.