Multimodal synaptomics reveals coordinated synaptic activity, protein synthesis and reorganization in NMDAR regulation

Complex neuronal circuit functions emerge from local, actively regulated synaptic protein levels that interplay with synaptic neurotransmission across heterogenous synapse populations. Understanding the mechanisms by which chemical and disease-associated genetic perturbations impact neuronal circuit functions requires simultaneous measurement of these factors with single-synapse resolution at population scale. Here, we combine in situ multimodal imaging of local mRNA translation, synaptic multiprotein composition, and synapse activity measured via calcium or glutamate fluxes, within the same spatially resolved synapses. We apply this approach of multimodal synapse profiling to study ketamine plasticity. Results map a causal network of NR2A-depletion-induced changes to synaptic scaffolding and receptor proteins, driven by synaptic activity and local mRNA translation, which translates to Grin2a models of schizophrenia in vitro and in vivo . Thus, multimodal synaptomics can reveal mechanistic neurobiology that underlies chemical and genetic perturbations within the context of scalable neuronal cultures, which can serve as models for human disease and therapeutic development.