Abstract
The physical properties of the postsynaptic membrane (PSM), including its viscosity, determine its capacity to regulate the net flux of synaptic membrane proteins such as neurotransmitter receptors. To address these properties, we studied the lateral diffusion of glycophosphatidylinositol-anchored green fluorescent protein and cholera toxin bound to the external leaflet of the plasma membrane. Relative to extrasynaptic regions, their mobility was reduced at synapses and even more at inhibitory than at excitatory ones. This indicates a higher density of obstacles and/or higher membrane viscosity at inhibitory contacts. Actin depolymerization reduced the confinement and accelerated a population of fast, mobile molecules. The compaction of obstacles thus depends on actin cytoskeleton integrity. Cholesterol depletion increased the mobility of the slow diffusing molecules, allowing them to diffuse more rapidly through the crowded PSM. Thus, the PSM has lipid-raft properties, and the density of obstacles to diffusion depends on filamentous actin. Therefore, lipid composition and actin-dependent protein compaction regulate viscosity of the PSM and, consequently, the molecular flow in and out of synapses.