Mixed-culture assays for analyzing neuronal synapse formation

被引:118
作者
Biederer, Thomas [1 ]
Scheiffele, Peter [2 ]
机构
[1] Yale Univ, Dept Mol Biophys & Biochem, New Haven, CT 06520 USA
[2] Columbia Univ, Ctr Neurobiol & Behav, Dept Physiol & Cellular Biophys, New York, NY 10032 USA
关键词
D O I
10.1038/nprot.2007.92
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:670 / 676
页数:7
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