The molecular mechanisms underlying MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)-induced dopaminergic (DAergic) neuronal death in vivo are still not fully understood. To investigate the selective DAergic neurotoxicity, we have developed an immunological technique to isolate DAergic synaptosomes from mouse striatal tissues using an antibody against 20 amino acid residues in the extracellular second loop of dopamine transporter (DAT). The DAT protein level in the isolated DAergic synaptosomes was markedly decreased at 16 h after a single injection of 30 mg/kg MPTP, but not in striatal homogenate and crude synaptosomes fraction. GBR-12909, a dopamine uptake inhibitor, completely reversed the MPTP-induced decrease of DAT protein in the DAergic synaptosomes. These results suggest that the isolated DAergic synaptosomes can be useful to identify mechanisms of loss of the nerve terminals.
Copyright © 2010 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.