The brain is characterized by an enormous degree of complexity and diversity of neural networks, making it one of the most complicated organs, and plays essential roles in regulating body and mind, maintaining thinking and other important physiology actions. This optimized method is simpler and easier to perform than the traditional method for the examination of the synaptogenic activities of individual cell-surface proteins in isolation, and provides an unbiased screening platform for synaptogenic proteins. This protocol describes a new co-culture method in which cDNA plasmid is transfected into human embryonic kidney 293T cells using polyetherimide 24 h after cells were mixed with neurons, and immunostaining and confocal imaging are employed for analyzing synaptogenesis. However, controlling a large number of cell pools in co-culture is complicated, creating a potential barrier for high-throughput screening. The artificial synapse formation assay, in which non-neuronal cells and neurons are co-cultured, has been shown to be a powerful system for screening CAMs.

Synaptic cell adhesion molecules (CAMs) are thought to play essential roles in the initiation of the synapse formation process. The proper formation of synapses is essential for brain function.

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