C. Saunders et al., Amphetamine-induced loss of human dopamine transporter activity: An internalization-dependent and cocaine-sensitive mechanism, P NAS US, 97(12), 2000, pp. 6850-6855
Citations number
40
Categorie Soggetti
Multidisciplinary
Journal title
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
The dopamine transporter (DAT) is a target of amphetamine (AMPH) and cocain
e. These psychostimulants attenuate DAT clearance efficiency, thereby incre
asing synaptic dopamine (DA) levels. Re-uptake rate is determined by the nu
mber of functional transporters at the cell surface as well as by their tur
nover rate. Here, we present evidence that DAT substrates, including AMPH a
nd DA, cause internalization of human DAT, thereby reducing transport capac
ity. Acute treatment with AMPH reduced the maximal rate of [H-3]DA uptake,
decreased AMPH-induced currents, and significantly redistributed the immuno
fluorescence of an epitope-tagged DAT from the plasma membrane to the cytos
ol in human embryonic: kidney 293 cells. Conversely, DAT inhibitors, such a
s cocaine, mazindol, and nomifensine, when administered with AMPH, blocked
the reduction in [H-3]DA uptake and the redistribution of DAT immunofluores
cence to the cytosol. The reductions of [H-3]DA uptake and AMPH-induced DAT
internalization also were inhibited by coexpression of a dominant negative
mutant of dynamin I (K44A), indicating that endocytosis modulates transpor
t capacity, likely through a clathrin-mediated pathway. With this mechanism
of regulation, acute application of AMPH would reduce DA uptake not only b
y direct competition for uptake, but also by reducing the available cell-su
rface DAT. Moreover, AMPH-induced internalization might diminish the amount
of DAT available for DA efflux, thereby modulating the cytotoxic effects o
f elevated extracellular DA.