Pyk2 Overexpression in Postsynaptic Neurons Blocks Aβ 1-42 -induced Synaptotoxicity in a Microfluidic Co-Culture Model

Recent meta-analyses of genome-wide association studies identified a number of genetic risk factors of Alzheimer’s disease (AD); however, little is known about the mechanisms by which they contribute to the pathological process. As synapse loss is observed at the earliest stage of AD, deciphering the impact of AD risk genes on synapse formation and maintenance is of great interest. In this paper, we report a microfluidic co-culture device that physically isolates synapses from pre- and postsynaptic neurons and chronically exposes them to toxic amyloid-beta (Aβ) peptides secreted by model cell lines overexpressing wild-type or mutated (V717I) amyloid precursor protein (APP). Co-culture with cells overexpressing mutated APP exposed the synapses of primary hippocampal neurons to Aβ molecules at nanomolar concentrations and induced a significant decrease in synaptic connectivity, as evidenced by distance-based assignment of postsynaptic puncta to presynaptic puncta. Treating the cells with antibodies that target different forms of Aβ suggested that low molecular weight oligomers are the likely culprit. As proof of concept, we demonstrate that overexpression of Pyk2 –an AD risk gene involved in synaptic plasticity and shown to decrease in AD brains at gene expression and protein levels– selectively in postsynaptic neurons is protective against Aβ-induced synaptotoxicity. In summary, our lab-on-a-chip device provides a physiologically-relevant model of AD-related synaptotoxicity, optimal for assessing the impact of risk genes in pre- and postsynaptic compartments.