Mixed-culture assays for analyzing neuronal synapse formation
Abstract
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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Heterologous expression of key synaptogenic proteins in non-neuronal cells allows them to receive projections from neurons and subsequently induce morphological and functional presynaptic differentiation in the contacting axons.
“It has been shown previously that heterologous expression of key synaptogenic proteins in non-neuronal cells allows them to receive projections from neurons and subsequently induce morphological and functional presynaptic differentiation in the contacting axons (Biederer and Scheiffele, 2007).”
From Comprehensive expression analyses of neural cell-type-specific miRNAs identify new determinants of the specification and maintenance of neuronal phenotypes (2013) · cites this paper as Biederer and Scheiffele, 2007
Neuroligin 1 possesses synaptogenic activity.
“Neuroligin 1 is among the neuronal proteins to possess such synaptogenic activity (Scheiffele et al., 2000;Biederer and Scheiffele, 2007)”
From Comprehensive expression analyses of neural cell-type-specific miRNAs identify new determinants of the specification and maintenance of neuronal phenotypes (2013) · cites this paper as Biederer and Scheiffele, 2007
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Self-citationYamagata, M., Sanes, J. R., & Weiner, J. A. (2003) · Current Opinion in Cell Biology · doi:10.1016/s0955-0674(03)00107-8 · Cited by 374 · see citing works
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Dual proteome-scale networks reveal cell-specific remodeling of the human interactome
Huttlin, E. L., Bruckner, R. J., Navarrete-Perea, J., Cannon, J. R., Baltier, K., Gebreab, F., Gygi, M. P., Thornock, A., Zarraga, G., Tam, S., et al. (2020) · doi:10.1101/2020.01.19.905109 · Cited by 960 · see citing works
Truncating mutations in NRXN2 and NRXN1 in autism spectrum disorders and schizophrenia
Gauthier, J., Siddiqui, T. J., Huashan, P., Yokomaku, D., Hamdan, F. F., Champagne, N., Lapointe, M., Spiegelman, D., Noreau, A., Lafrenière, R. G., et al. (2011) · Human Genetics · doi:10.1007/s00439-011-0975-z · Cited by 286 · see citing works
Protein tyrosine phosphatases PTPδ, PTPσ, and LAR: presynaptic hubs for synapse organization
Takahashi, H., & Craig, A. M. (2013) · Trends in Neurosciences · doi:10.1016/j.tins.2013.06.002 · Cited by 257 · see citing works
Jovičić, A., Roshan, R., Moisoi, N., Pradervand, S., Moser, R., Pillai, B., & Luthi-Carter, R. (2013) · The Journal of Neuroscience · doi:10.1523/jneurosci.0600-12.2013 · Cited by 251 · see citing works
Elfn1 Regulates Target-Specific Release Probability at CA1-Interneuron Synapses
Sylwestrak, E. L., & Ghosh, A. (2012) · Science · doi:10.1126/science.1222482 · Cited by 151 · see citing works
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Pooled genetic perturbation screens with image-based phenotypes
Feldman, D., Funk, L., Le, A., Carlson, R. J., Leiken, M. D., Tsai, F., Soong, B., Singh, A., & Blainey, P. C. (2022) · Nature Protocols · doi:10.1038/s41596-021-00653-8 · Cited by 59 · see citing works
- Authors
- Biederer, T., & Scheiffele, P.
- Venue
- Nature Protocols
- Published
- 2007-03
- Volume
- 2
- Issue
- 3
- Pages
- 670-676
- ISSN
- 1754-2189
- 1750-2799
- Cited by
- 143
- References
- 2
- Language
- en
- Licence
- www.springer.com/tdm (tdm)
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