Isolation of Synaptic Vesicles

Synaptic vesicles are the most abundant secretory organelle in eukaryotic neural cells. Synaptic vesicles are physically distinct from other membrane-bound organelles because they are small, spherical, and highly uniform in size with a diameter of about 40 nm. Synaptic vesicles also have a characteristically low density because water and phospholipids account for 88% of an individual synaptic vesicle's weight. The homogeneous size and density of the synaptic vesicle are characteristics that are exploited in most synaptic vesicle isolation procedures. Synaptic vesicles are purified by isopycnic/velocity sedimentation and size-based purification schemes. However, protocols differ in the tissue source of vesicles, the way the tissue is homogenized, and the way the vesicles are fractionated. This unit describes two well-characterized and widely used synaptic vesicle isolation procedures that are capable of generating membrane fractions that are 20 to 30 times enriched in synaptic vesicles.