Journal
NATURE PROTOCOLS
Volume 2, Issue 3, Pages 670-676Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2007.92
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Funding
- NIDA NIH HHS [R01 DA018928] Funding Source: Medline
- NINDS NIH HHS [R01 NS532830] Funding Source: Medline
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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.
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