A rapid method for preparing synaptosomes: comparison, with alternative procedures.
A method for the rapid (1-1.5 h) preparation of nerve ending particles (synaptosomes) from rat cerebral cortex is described. The synaptosome fraction has been characterized by quantitative electron microscopy and enzyme distribution studies. By these criteria, the fraction showed a degree of enrichment in synaptic structures which was comparable to that of the standard (4-5 h) preparation, and substantially better than an alternative fast (2-2.5 h) method. On incubation, synaptosomes obtained by the new procedure accumulated a high tissue concentration of potassium and showed a high, linear rate of oxygen uptake. Depolarization by veratrine caused a significant increase in the rate of respiration and in the release of the physiologically active amino acids; glutamate, aspartate and GABA, as well as a significant reduction in tissue potassium. Thus, the new procedure compared favourably with alternative methods as judged by these indices of metabolic and functional performance. The new preparation method has been found to be of value in metabolic studies of synaptosomes prepared from human post-mortem brain.
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002540:Cerebral Cortex / Q000166:cytology*; D004798:Enzymes / Q000378:metabolism; D005260:Female; D008297:Male; D008854:Microscopy, Electron; D010101:Oxygen Consumption; D051381:Rats; D011919:Rats, Inbred Strains; D013574:Synaptosomes / Q000201:enzymology / Q000648:ultrastructure*
D000596:Amino Acids; D004798:Enzymes