Imaging synaptogenesis by measuring accumulation of synaptic proteins

Synapse formation and modification involve the successive recruitment of pre- and post-synaptic signaling molecules. The procedure presented here investigates the function of specific synaptic factors in gain- and loss-of-function experiments by measuring protein accumulation at synapses using quantitative immunohistochemistry, confocal microscopy, and image analysis. Although the procedure is described for dissociated cultures of hippocampal neurons, it can be extended to various neuronal cell types in culture.