Probing Rotational Viscosity in Synaptic Vesicles

The synaptic vesicle (SV) is a central organelle in neurotransmission, and previous studies have suggested that SV protein 2 (SV2) may be responsible for forming a gel-like matrix within the vesicle. Here we measured the steady-state rotational anisotropy of the fluorescent dye, Oregon Green, within individual SVs. By also measuring the fluorescence lifetime of Oregon Green in SVs, we determined the mean rotational viscosity to be 16.49 +/- 0.12 cP for wild-type (WT) empty mice vesicles (i.e., with no neurotransmitters), 11.21 +/- 0.12 cP for empty vesicles from SV2 knock-out mice, and 11.40 +/- 0.65 cP for WT mice vesicles loaded with the neurotransmitter glutamate (Glu). This measurement shows that SV2 is an important determinant of viscosity within the vesicle lumen, and that the viscosity decreases when the vesicles are filled with Glu. The viscosities of both empty SV2 knock-out vesicles and Glu-loaded WT vesicles were significantly different from that of empty WT SVs (p < 0.05). This measurement represents the smallest enclosed volume in which rotational viscosity has been measured thus far.