title: Phorbol esters increase dopamine transporter phosphorylation and decrease transport Vmax.
abstract: Sodium- and chloride-coupled transport of dopamine from synapses into presynaptic terminals plays a key role in terminating dopaminergic neurotransmission. Regulation of the function of the dopamine transporter, the molecule responsible for this translocation, is thus of interest. The primary sequence of the dopamine transporter contains multiple potential phosphorylation sites, suggesting that the function of the transporter could be regulated by phosphorylation. Previous work from this laboratory has documented that phorbol ester activation of protein kinase C (PKC) decreases dopamine transport Vmax in transiently expressing COS cells. In the present report, we document in vivo phosphorylation of the rat dopamine transporter stably expressed in LLC-PK1, cells and show that phosphorylation is increased threefold by phorbol esters. Dopamine uptake is also regulated by phorbol esters in these cells; phorbol 12-myristate 13-acetate (PMA) reduces transport Vmax by 35%. Parallels between the time course, concentration dependency, and staurosporine sensitivity of alterations in transporter phosphorylation and transporter Vmax suggest that dopamine transporter phosphorylation involving PKC could contribute to this decreased transporter function. Phosphorylation of the dopamine transporter by PKC or by a PKC-activated kinase could be involved in rapid neuroadaptive processes in dopaminergic neurons.
authors.affiliation: Molecular Neurobiology Branch, National Institute on Drug Abuse, Baltimore, MD 21224, USA.
authors.forename: R A
authors.initials: RA
authors.lastname: Huff
authors.forename: R A
authors.initials: RA
authors.lastname: Vaughan
authors.forename: M J
authors.initials: MJ
authors.lastname: Kuhar
authors.forename: G R
authors.initials: GR
authors.lastname: Uhl
mesh_terms: D000409:Alanine / Q000493:pharmacokinetics; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000378:metabolism*; D004298:Dopamine / Q000493:pharmacokinetics*; D050483:Dopamine Plasma Membrane Transport Proteins; D018374:LLC-PK1 Cells; D007930:Leucine / Q000493:pharmacokinetics; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
chemical_list: D002352:Carrier Proteins; D050483:Dopamine Plasma Membrane Transport Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D007930:Leucine; D013755:Tetradecanoylphorbol Acetate; D000409:Alanine; D004298:Dopamine