Mitogen-activated protein kinase regulates dopamine transporter surface expression and dopamine transport capacity

José A. Morón, Irina Zakharova, Jasmine V. Ferrer, Gerald A. Merrill, Bruce Hope, Eileen M. Lafer, Zhi Cheng Lin, Jia Bei Wang, Jonathan A. Javitch, Aurelio Galli, Toni S. Shippenberg

Research output: Contribution to journalArticle

194 Scopus citations

Abstract

The dopamine transporter (DAT) regulates the clearance of dopamine (DA) released into the extracellular space and is an important site on which psychostimulants act to produce their effects. Here, we show that mitogen-activated protein kinase (MAPK) regulates the transport capacity and intracellular trafficking of DAT. Incubation of striatal synaptosomes or epitope-tagged human DAT (hDAT) human embryonic kidney (HEK) 293 cells with the MAPK kinase (MEK) inhibitors 1,4-diamino-2,3-dicyano-1,4-bis(o-aminophenylmercapto) butadiene and 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one decreased DA uptake in a concentration- and time-dependent manner. Kinetic studies revealed a decrease in the capacity of transport (Vmax but no change in Km. Immunoblotting confirmed labeling of p42 and p44 MAPK in untreated striatal synaptosomes and HEK 293 cells, consistent with constitutive MAPK activation, and the inhibitors used decreased MAPK phosphorylation. Biotinylation and confocal imaging studies showed that MAPK inhibition promoted the clathrin-associated redistribution of hDAT from the plasma membrane to the cytosol. In contrast, transient transfection of hDAT-expressing cells with constitutively active MEK increased the Vmax of DA transport without altering Km. However, only a small increase in hDAT cell surface expression was seen. These data demonstrate an involvement of the MAPK cascade in regulating DAT transport capacity in striatum and that inhibition of this cascade decreases DAT cell surface expression in HEK 293 cells. Furthermore, they highlight the potential role of MAPK as a presynaptic mechanism that regulates DA signaling.

Original languageEnglish
Pages (from-to)8480-8488
Number of pages9
JournalJournal of Neuroscience
Volume23
Issue number24
StatePublished - Sep 17 2003
Externally publishedYes

Keywords

  • Biotinylation
  • Dopamine transporter
  • Epitope-tagged-hDAT
  • HEK 293 cells
  • MAPK
  • Trafficking

Fingerprint Dive into the research topics of 'Mitogen-activated protein kinase regulates dopamine transporter surface expression and dopamine transport capacity'. Together they form a unique fingerprint.

  • Cite this

    Morón, J. A., Zakharova, I., Ferrer, J. V., Merrill, G. A., Hope, B., Lafer, E. M., Lin, Z. C., Wang, J. B., Javitch, J. A., Galli, A., & Shippenberg, T. S. (2003). Mitogen-activated protein kinase regulates dopamine transporter surface expression and dopamine transport capacity. Journal of Neuroscience, 23(24), 8480-8488.