Institute for Cell Biology and Neuroscience, Biologicum, Goethe-University, Frankfurt am Main, Germany.
Institute of Pharmaceutical Chemistry, Cluster of Excellence "Macromolecular Complexes", Goethe-University, Frankfurt am Main, Germany.
Mol Cell Neurosci. 2014 Mar;59:106-18. doi: 10.1016/j.mcn.2014.02.003. Epub 2014 Feb 15.
Neurotransmitter release as well as the structural and functional dynamics of the presynaptic active zone is controlled by proteinaceous components. Here we describe for the first time an experimental approach for the isolation of the presynaptic active zone from individual mouse brains, a prerequisite for understanding the functional inventory of the presynaptic protein network and for the later analysis of changes occurring in mutant mice. Using a monoclonal antibody against the ubiquitous synaptic vesicle protein SV2 we immunopurified synaptic vesicles docked to the presynaptic plasma membrane. Enrichment studies by means of Western blot analysis and mass spectrometry identified 485 proteins belonging to an impressive variety of functional categories. Our data suggest that presynaptic active zones represent focal hot spots that are not only involved in the regulation of neurotransmitter release but also in multiple structural and functional alterations the adult nerve terminal undergoes during neural activity in adult CNS. They furthermore open new avenues for characterizing alterations in the active zone proteome of mutant mice and their corresponding controls, including the various mouse models of neurological diseases.
神经递质的释放以及突触前活性区的结构和功能动态受蛋白质成分的控制。在这里，我们首次描述了一种从单个小鼠大脑中分离突触前活性区的实验方法，这是理解突触前蛋白网络功能清单以及分析突变小鼠中发生变化的前提条件。我们使用一种针对普遍存在的突触小泡蛋白 SV2 的单克隆抗体，免疫纯化与突触前质膜对接的突触小泡。通过 Western blot 分析和质谱分析的富集研究，鉴定出属于多种功能类别的 485 种蛋白质。我们的数据表明，突触前活性区代表焦点热点，不仅参与神经递质释放的调节，还参与成年中枢神经系统神经活动过程中成年神经末梢经历的多种结构和功能改变。它们还为描述突变小鼠和相应对照的活性区蛋白质组的改变开辟了新途径，包括各种神经疾病的小鼠模型。
