Separation of synaptic junctional complexes from thin layers of rabbit cerebral cortex.
Synaptic junctional fractions were separated from rabbit brain by procedures based on combining the methods of Cotman and Taylor [4], Orosz et al. [16, 17] and Lisman et al. [13]. Thin layers of cerebral cortices were homogenized to obtain a crude mitochondrial-synaptosomal fraction. The sedimentation rates of mitochondria and mitochondria containing synaptosomes were increased by raising the density of mitochondria with an insoluble dense formazan deposit inside mitochondria after iodo-nitrotetrazolium treatment. The synaptic plasma membrane fraction isolated by this method contained no mitochondrial contamination. After Triton X-100 treatment the insoluble residues of the detergent were centrifuged through discontinuous sucrose gradients. A great enrichment of morphologically identifiable intact synaptic junctions was observed in some of the obtained interface layers.
D000818:Animals; D002500:Centrifugation, Isopycnic / Q000379:methods; D002540:Cerebral Cortex / Q000032:analysis* / Q000648:ultrastructure; D003902:Detergents; D008928:Mitochondria / Q000032:analysis* / Q000648:ultrastructure; D017830:Octoxynol; D011092:Polyethylene Glycols; D011817:Rabbits; D011869:Radioligand Assay; D013570:Synaptic Membranes / Q000032:analysis* / Q000648:ultrastructure
D003902:Detergents; D011092:Polyethylene Glycols; D017830:Octoxynol