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Neuron-fibroblast coculture assay to demonstrate presynaptic differentiation induced by the NGL family of cell adhesion molecules

External protocol Created on 03 May 2014

Authors

Seho Kim and Eunjoon Kim

Summary

Expression of synaptic cell adhesion molecules, such as neuroligin, neurexin and SynCAM, in nonneural cells can induce pre- or postsynaptic differentiation in contacting neurites of cocultured neurons [1-3]. We used this assay to demonstrate that NGL, a family of cell adhesion molecules that associates with the netrin-G family of cell adhesion molecules and the postsynaptic scaffolding protein PSD-95, can induce presynaptic differentiation in contacting axons of cocultured neurons. Presynaptic differentiation was visualized by immunostaining for the presynaptic marker synapsin I. In addition, functional presynaptic differentiation was demonstrated by vesicle turnover experiments, which monitor the uptake of synaptotagmin luminal domain antibodies [4].

Further details

The protocol was published on Protocol Exchange in 2006. To see the entire protocol, click on the source link.

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