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Volume 17, Number 3,
Issue of February 1, 1997
pp. 960-974
Copyright ©1997 Society for Neuroscience
Multiple Ionic Conductances of the Human Dopamine Transporter:
The Actions of Dopamine and Psychostimulants
Received Aug. 29, 1996; revised Nov. 12, 1996; accepted Nov. 18, 1996.
Mark S. Sonders1, 2,
Si-Jia Zhu3,
Nancy R. Zahniser3,
Michael P. Kavanaugh2, and
Susan G. Amara1, 2
1 The Howard Hughes Medical Institute and
2 Vollum Institute, Oregon Health Sciences University,
Portland, Oregon 97201, and 3 Department of Pharmacology,
University of Colorado Health Sciences Center, Denver, Colorado 80262
Electrophysiological and pharmacological studies of a cloned human
dopamine transporter (hDAT) were undertaken to investigate the
mechanisms of transporter function and the actions of drugs at this
target. Using two-electrode voltage-clamp techniques with hDAT-expressing Xenopus laevis oocytes, we show that
hDAT can be considered electrogenic by two criteria. (1) Uptake of hDAT substrates gives rise to a pharmacologically appropriate
"transport-associated" current. (2) The velocity of DA uptake
measured in oocytes clamped at various membrane potentials was
voltage-dependent, increasing with hyperpolarization. Concurrent
measurement of transport-associated current and substrate flux in
individual oocytes revealed that charge movement during substrate
translocation was greater than would be expected for a transport
mechanism with fixed stoichiometry of 2 Na+ and 1 Cl per DA+ molecule. In addition to the
transport-associated current, hDAT also mediates a constitutive leak
current, the voltage and ionic dependencies of which differ markedly
from those of the transport-associated current. Ion substitution
experiments suggest that alkali cations and protons are carried by the
hDAT leak conductance. In contrast to the transport-associated
functions, the leak does not require Na+ or
Cl, and DAT ligands readily interact with the transporter
even in the absence of these ions. The currents that hDAT mediates
provide a functional assay that readily distinguishes the modes of
action of amphetamine-like "DA-releasing" drugs from cocaine-like
translocation blockers. In addition, the voltage dependence of DA
uptake suggests a mechanism through which presynaptic DA autoreceptor
activation may accelerate the termination of dopaminergic
neurotransmission in vivo.
Key words:
Na+/Cl-dependent;
carrier;
cocaine;
amphetamine;
methamphetamine;
methylphenidate;
MPP+;
uptake;
release;
Xenopus oocyte;
psychomotor stimulant
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International Union of Pharmacology. XIX. The IUPHAR Receptor Code: A Proposal for an Alphanumeric Classification System
Pharmacol. Rev.,
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[Abstract]
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S. R. Jones, R. R. Gainetdinov, R. M. Wightman, and M. G. Caron
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J. Neurosci.,
March 15, 1998;
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[Abstract]
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S.-J. Zhu, M. P. Kavanaugh, M. S. Sonders, S. G. Amara, and N. R. Zahniser
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[Abstract]
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A. Galli, C.I. Petersen, M. deBlaquiere, R.D. Blakely, and L.J. DeFelice
Drosophila Serotonin Transporters Have Voltage-Dependent Uptake Coupled to a Serotonin-Gated Ion Channel
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[Abstract]
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Y. Cao, S. Mager, and H. A. Lester
H+ Permeation and pH Regulation at a Mammalian Serotonin Transporter
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[Abstract]
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J. Lotharius and K. L. O'Malley
The Parkinsonism-inducing Drug 1-Methyl-4-phenylpyridinium Triggers Intracellular Dopamine Oxidation. A NOVEL MECHANISM OF TOXICITY
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[Abstract]
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M. Quick and B. R. Stevens
Amino Acid Transporter CAATCH1 Is Also an Amino Acid-gated Cation Channel
J. Biol. Chem.,
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[Abstract]
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Y. G. Ni, J.-G. Chen, A. Androutsellis-Theotokis, C.-J. Huang, E. Moczydlowski, and G. Rudnick
A Lithium-induced Conformational Change in Serotonin Transporter Alters Cocaine Binding, Ion Conductance, and Reactivity of Cys-109
J. Biol. Chem.,
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[Abstract]
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N. MacAulay, A. Bendahan, C. J. Loland, T. Zeuthen, B. I. Kanner, and U. Gether
Engineered Zn2+ Switches in the gamma -Aminobutyric Acid (GABA) Transporter-1. DIFFERENTIAL EFFECTS ON GABA UPTAKE AND CURRENTS
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[Abstract]
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D. H. Feldman, W. R. Harvey, and B. R. Stevens
A Novel Electrogenic Amino Acid Transporter Is Activated by K+ or Na+, Is Alkaline pH-dependent, and Is Cl--independent
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[Abstract]
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