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Research Summary

Multiplexed super-resolution imaging of synaptic nanoarchitecture

Regulated neuronal transmission implies a very precise ultrastructural organization of the pre- and postsynapse. Although most molecular players of the synapse are known, their exact nanoscopic arrangement still remains an unsolved puzzle. Our goal is to resolve the molecular makeup and the nanoarchitechture of synaptic specializations, including pools of synaptic vesicles, the vesicle release site (active zone – AZ), and the neurotransmitter reception apparatus (postsynaptic density – PSD). To investigate the precise arrangement of synaptic proteins relative to each other, we employ a multiplex super-resolution approach. Our fully automated 3D direct stochastic optical reconstruction microscopy (dSTORM) setup equipped with a pipetting system enables the acquisition of >15 target proteins at individual synapses with <10 nm localization precision. As our main model system, we use the calyx of Held, a biophysically well-studied giant glutamatergic synapse of the mammalian central nervous system with a well-characterized geometry. Complementing multiplex super-resolution by other approaches available in our laboratory such as electron microscopy, viral gene transfer, and electrophysiology we aim to create a nanoscale molecular map of a glutamatergic synapse and derive functional principles.