Charting the nanotopography of inner hair cell synapses using MINFLUX nanoscopy

For us to hear, the cochlea encodes sounds into neural signals at synapses of inner hair cells (IHCs) and the auditory nerve with remarkable fidelity. To achieve the high rates of temporally precise synaptic transmission over long periods of time, IHCs employ sophisticated ribbon-type active zones (AZ). In order for us to understand synaptic sound encoding, we need to decipher the underpinning molecular topography of these synapse which had remained challenging due to technological limitations. In our study, we applied 3-dimensional minimal flux optical nanoscopy (3D MINFLUX) to mouse IHC synapses to chart the position of key pre- and postsynaptic proteins with single digit nanometre resolution of imaging. We demonstrate that nanoclusters of channels and interacting proteins govern the topography of AZs and postsynaptic densities (PSDs). We count synaptic proteins, their nanoclusters and determine their spatial organization feeding into computational modelling of AZ function. In conclus..., , # Charting the nanotopography of inner hair cell synapses using MINFLUX nanoscopy Dataset DOI: [10.5061/dryad.qfttdz0v3](10.5061/dryad.qfttdz0v3) ## Description of the data and file structure This dataset was generated for the study *“Charting the nanotopography of inner hair cell synapses using MINFLUX nanoscopy”*, which aimed to elucidate the three-dimensional nanoscale organisation of key synaptic proteins at inner hair cell (IHC) ribbon synapses. Using MINFLUX nanoscopy, we acquired high-precision localisations to chart the organisation of key pre- and post-synaptic proteins, including RIBEYE, RBP2, CaV1.3, Piccolino, Homer1, GluA2, and VGLUT3 to generate a molecular atlas of these specialised sensory synapses. The data was processed to extract molecular coordinates, analyse spatial clustering patterns, and derive quantitative descriptors of synaptic architecture and nanocluster organisation. ### Files and variables #### File: Alpha_shape_volume.xlsx **Description:** Volume of...,