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Volume 17, Number 14, Issue of July 15, 1997 pp. 5255-5262
Copyright ©1997 Society for Neuroscience

Immunogold Localization of the Dopamine Transporter: An Ultrastructural Study of the Rat Ventral Tegmental Area

Received Dec. 10, 1996; revised March 11, 1997; accepted March 13, 1997.

Melissa J. Nirenberg1, June Chan1, Roxanne A. Vaughan2, George R. Uhl3, 4, Michael J. Kuhar5, and Virginia M. Pickel1

1 Department of Neurology and Neuroscience, Cornell University Medical College, New York, New York 10021, Branches of 2 Neuroscience and 3 Molecular Neurobiology, National Institute on Drug Abuse, Baltimore, Maryland 21224, 4 Departments of Neurology and Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21224, and 5 Neuroscience Division, Yerkes Regional Primate Center, Emory University, Atlanta, Georgia 30322

The dopamine transporter (DAT) plays an important role in the plasmalemmal reuptake of dopamine and, thus, in the termination of normal dopaminergic neurotransmission. DAT is also a major binding site for cocaine and other stimulants, the psychoactive effects of which are associated primarily with the inhibition of dopamine reuptake within mesocorticolimbic dopaminergic neurons. We used electron microscopy with an anti-peptide antiserum directed against the N-terminal domain of DAT to determine the subcellular localization of this transporter in the rat ventral tegmental area (VTA), the region that contains the cell bodies and dendrites of these dopaminergic neurons. We show that in the VTA, almost 95% of the DAT immunogold-labeled profiles are neuronal perikarya and dendrites, and the remainder are unmyelinated axons. Within perikarya and large proximal dendrites, almost all of the DAT immunogold particles are associated with intracellular membranes, including saccules of Golgi and cytoplasmic tubulovesicles. In contrast, within medium- to small-diameter dendrites and unmyelinated axons, most of the DAT gold particles are located on plasma membranes. In dually labeled tissue, peroxidase reaction product for the catecholamine-synthesizing enzyme tyrosine hydroxylase is present in DAT-immunoreactive profiles. These findings suggest that intermediate and distal dendrites are both the primary sites of dopamine reuptake and the principal targets of cocaine and related psychostimulants within dopaminergic neurons in the VTA.

Key words: dopamine; transporter; uptake; ultrastructure; ventral tegmental area; midbrain; mesocorticolimbic; electron microscopy; immunogold; dendritic release; plasma membrane; cocaine; amphetamine; neurotoxicity




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