Subfractionation of Ficoll Gradient Purified Synaptosomes: GABA Uptake by the Subfractions

Abstract

The classical synaptosomes preparation purified on discontinuous Ficoll gradient has been sufractionated into three further subfractions by introducing more Ficoll density layers.

Among the three subfractions, the lightest one was the one which was more difficult to obtain in reproducible amounts and was only partially characterized in terms of labelled GABA uptake. The heaviest one most probably is largely made up of partially damaged nerve endings. The central one was the most reproducible in terms of yield and labelled GABA uptake and actually was the one we studied more thoroughly in terms of morphology, labelled GABA uptake and its pharmacology. A comparison has been made with the classical “total” purified synaptosomes fraction.

An interesting result of these experiments is a paradoxical effect of the glial uptake inhibitor β-alanine. This substance appears to favor, in both “total” and “central” fraction synaptosomes, a redistribution of taken up GABA from contaminating glia to actual nerve endings.

Keywords:

• Rat brain
• Synaptosomes
• Ficoll gradients
• Subfractionation
• GABA uptake