MACROMOLECULAR ARCHITECTURE OF THE SYNAPSE

突触的大分子结构

• 批准号: 6111979
• 负责人: Thomas S Reese
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6111979
• 关键词: actins; calmodulin dependent protein kinase; cell component structure /function; electron microscopy; freezing; glass; nerve /myelin protein; spectrin; structural biology; synapses

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.

这个新项目分析了分子体系结构 哺乳动物大脑中的突触，最初关注 突触后密度（PSD）。许多分子成分 PSD是已知的，但是PSD是如此巨大（大约10至 必须开发新方法以了解的第九个da） 如何组织其大量分子成分。一个 有希望的新方法首先将PSD连接到玻璃和 然后冷冻并复制它们以进行电子显微镜。 释放之前的操作允许识别和本地化 特定组成部分。其中包括：PSD的方向 在玻璃上通过先前用抗体的玻璃处理一种 它们的表面成分； PSD的部分消化 带洗涤剂的玻璃；和标签，以上任何一个之后 用胶体黄金标记的抗体。迄今为止工作清楚 区分PSD的裂缝，其中NR-1和PSD-95 从CaMKII所定位的细胞质侧进行了定位。 裂缝的消化揭示了形成的晶格状结构 PSD的核心；该晶格标记为PSD-95。这 Spect和psd的Spectin和Actin的分布目前正在 调查，Camkii的分布是 另一个项目。现在显而易见的是 PSD的组件是PSD函数的重要方面。