Nanoscale Subsynaptic Domains Underlie the Organization of the Inhibitory Synapse

Inhibitory synapses mediate the majority of synaptic inhibition in the brain, thereby controlling neuronal excitability, firing, and plasticity. Although essential for neuronal function, the central question of how these synapses are organized at the subsynaptic level remains unanswered. Here, we use three-dimensional (3D) super-resolution microscopy to image key components of the inhibitory postsynaptic domain and presynaptic terminal, revealing that inhibitory synapses are organized into nanoscale subsynaptic domains (SSDs) of the gephyrin scaffold, GABA(A)Rs and the active-zone protein Rab3-interacting molecule (RIM). Gephyrin SSDs cluster GABA(A)R SSDs, demonstrating nanoscale architectural interdependence between scaffold and receptor. GABA(A)R SSDs strongly associate with active-zone RIM SSDs, indicating an important role for GABA(A)R nanoscale organization near sites of GABA release. Finally, we find that in response to elevated activity, synapse growth is mediated by an increase in the number of postsynaptic SSDs, suggesting a modular mechanism for increasing inhibitory synaptic strength.