Influence of synapsin I on synaptic vesicles: an analysis by force-volume mode of the atomic force microscope and dynamic light scattering.

Synaptic vesicles (SVs) are small neuronal organelles that store neurotransmitters and release them by exocytosis into the synaptic cleft for signal transmission between nerve cells. They consist of a highly curved membrane composed of different lipids containing several proteins with specific functions. A family of abundant extrinsic SV proteins, the synapsins, interact with SV proteins and phospholipids and play an important role in the regulation of SV trafficking and stability. We investigated the interactions of one these proteins with the SV membrane using atomic force microscope and dynamic light scattering. We examined SVs isolated from rat forebrain both under native conditions and after depletion of endogenous synapsin I. We used the atomic force microscope in two modes: imaging mode for characterizing the shape and size of SVs, and force-volume mode for characterizing their stiffness. Synapsin-depleted SVs were larger in size and showed a higher tendency to aggregate than native vesicles, although their stiffness was not significantly different. Because synapsins are believed to cross-link SV to each other and to the actin cytoskeleton, we also measured the SV aggregation kinetics induced by synapsin I by dynamic light scattering and atomic force microscopy and found that the addition of synapsin I promotes a rapid aggregation of SVs. The data indicate that synapsin directly affects SV stability and aggregation state and support the physiological role of synapsins in the assembly and regulation of SV pools within nerve terminals.