Mixed-culture assays for analyzing neuronal synapse formation
Mixed-culture assays for analyzing neuronal synapse formation
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DOI：
10.1038/nprot.2007.92
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Scheiffele, Peter
中科院分区：
文献类型：
--
作者：
Biederer, Thomas;Scheiffele, Peter
脊椎动物中枢神经系统中突触的组装需要跨突触间隙的双向信号传导，这种信号传导指导突触前膜和突触后膜结构域的分化。生化和遗传学研究已经确定了几种定位于突触且可能参与组织突触结构的黏附分子和信号分子。由于在此过程中有大量独立的信号相互协作，理解单个蛋白质如何对突触组织起作用变得复杂。本实验方案描述了一种检测系统，该系统允许单独检测单个细胞表面蛋白质的促突触形成活性。除了原代神经元培养物的制备和生长所需的时间（6 - 14天）外，该检测的实施和分析速度很快，大约需要2天。利用该检测方法，近期的研究表明，单个突触黏附复合物能够在很大程度上指导突触分化，并为突触发生的细胞生物学机制提供了新的见解。
The assembly of synapses in the vertebrate central nervous system requires bidirectional signaling across the synaptic cleft that directs the differentiation of pre- and postsynaptic membrane domains. Biochemical and genetic studies have identified several adhesion and signaling molecules that localize to synapses and might participate in organizing synaptic structures. Understanding how individual proteins contribute to synaptic organization is complicated by the fact that there are significant numbers of separate signals that cooperate in this process. This protocol describes an assay system that permits examination of synaptogenic activities of individual cell-surface proteins in isolation. Besides the time needed for preparation and growth of primary neuronal cultures (6-14 days), the execution and analysis of the assay is rapid, requiring approximately 2 days. Using this assay, recent studies revealed that single synaptic adhesion complexes can direct a remarkable degree of synaptic differentiation and provided new insights into the cell biological mechanisms of synaptogenesis.