Partial separation of synaptosomes accumulating 4-aminobutyrate or glutamate by zonal centrifugation on a discontinuous sucrose gradient.
Rat cerebral cortex slices were incubated with 1muM [3H]4-aminobutyrate or [3H]L-glutamine. The subcellular distribution of the accumulated labelled substances were determined by fractionating the nuclei-free homogenates on a 5-step discontinuous sucrose density gradient in a B XIV zonal rotor. The gradient was designed to separate the synaptosomes into 3 subpopulations of increasing density. The patterns of distribution of [3H]4-aminobutyrate and [3H-a1L-glutamate in the three synaptosomal peaks were distinctly different. This indicates the presence of separate types of nerve ending accumulating these two potential neurotransmitters, which are known to be metabolically closely linked through the enzyme glutamate decarboxylase. Recentrifugation of the 3 synaptosomal peaks on flat step gradients in a swingout rotor did not result in any further enrichment in transmitter-specific synaptosomes.
D000613:Aminobutyrates / Q000378:metabolism*; D000818:Animals; D002458:Cell Fractionation; D002501:Centrifugation, Zonal; D002540:Cerebral Cortex / Q000648:ultrastructure*; D005968:Glutamate Decarboxylase / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism*; D066298:In Vitro Techniques; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D051381:Rats; D013574:Synaptosomes / Q000201:enzymology / Q000378:metabolism*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D000613:Aminobutyrates; D005971:Glutamates; D005680:gamma-Aminobutyric Acid; D007770:L-Lactate Dehydrogenase; D005968:Glutamate Decarboxylase