Quantification of morpho-functional synaptic connectivity in dissociated neuronal cultures

Most neurological disorders share a phenotype of synaptic dysfunction. High-content microscopy is indispensable in the drug discovery process but the sensitivity relies on the accuracy of mature synapse detection. We present a workflow whereby synchronous calcium bursting is measured in GCaMP6f-transduced neurons. The same cultures are then subjected to Proximity Ligation Assay (PLA) to label mature synapses, the neurite network and nuclei. The integration of functional and morphological information provides a rich fingerprint of neuronal connectivity, deployable in different experimental conditions.