Evaluation of GABA uptake in subcellular fractions of bovine frontal cortex and brainstem

Abstract

GABA uptake as well as the distribution of GAT-1, GAT-2 and GAT-3 were investigated in bovine brain membrane fractions. GABA uptake was characterised by kinetic constants and IC50-values for a series of known inhibitors in subcellular fractions of frontal cortex and brainstem obtained by subsequent centrifugations on sucrose gradients. Additionally, the immunoreactivity for rGAT-1, rGAT-2 and rGAT-3 antibodies was studied in these fractions. The pharmacological profile for GABA uptake inhibition as well as results from immunoblotting indicated that GABA uptake in a selected subcellular fraction of frontal cortex (P2B) is almost exclusively due to GAT-1 whereas GABA uptake performed with a selected subcellular fraction of brainstem (P2A) in the presence of NNC 711 is mainly attributable to GAT-3.