Structural destabilization of synaptosomal particles by lysis and sequential chemical treatments

Synaptosomes from rat brains were subjected to a sequence of treatments: osmotic lysis, buffered saline wash, nonionic detergent, EGTA and EDTA. After each treatment, particulate samples were fixed in 2% glutaraldehyde-1% formaldehyde and centrifuged to form pellets which were then processed for and examined by electron microscopy. Five morphological classes of synaptic particle were defined in terms of character and presence of synaptic vesicles, flocculent and stranded material, designated as intervesicular scaffolding (IVS), and presynaptic membrane. During osmotic lysis, the presynaptic compartment was altered by loss of most, but not all, small synaptic vesicles, by increase in proportion of large vesicles, and by disappearance of the presynaptic densities. The retention of vesicles was interpreted in terms of IVS struts interconnecting anchorage sites on synaptic vesicles and the presynaptic junctional membrane. Treatment of lysed synaptosomes with nonionic detergent or EGTA resulted in loss of vesicles and IVS from the junctional region in most particles. The apposition of pre-and postsynaptic junctional membranes along the synaptic cleft was disrupted more by EGTA than by detergent. The final result of the sequential treatments was a sediment containing a high proportion of synaptic particles, about half of which had lost their presynaptic junctional membranes.