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Bead aggregation 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

The synaptic cell adhesion molecule neuroligin bound to beads has been shown to induce the clustering of neurxin and synaptic vesicle proteins in contacting axons [1]. In addition, neurexin-coated beads induced clustering of neuroligin and postsynaptic proteins in contacting dendrites [2]. We employed this assay to demonstrate that the extracellular domain of 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, bound to beads induces presynaptic differentiation in contacting axons of cultured neurons. Presynaptic differentiation was visualized by immunostaining for the presynaptic vesicle protein synaptophysin and vesicular glutamate transporter 1 (VGlut1; a marker for excitatory presynapses). In addition, functional presynaptic differentiation was demonstrated by vesicle turnover experiments, which monitor the uptake of synaptotagmin luminal domain antibodies [3].

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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