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Phorbol esters increase dopamine transporter phosphorylation and decrease transport Vmax

R A Huff1, R A Vaughan, M J Kuhar

  • 1Molecular Neurobiology Branch, National Institute on Drug Abuse, Baltimore, MD 21224, USA.

Journal of Neurochemistry
|January 1, 1997
PubMed
Summary
This summary is machine-generated.

Protein kinase C (PKC) activation phosphorylates the dopamine transporter, reducing its function. This phosphorylation may play a role in neuroadaptation within dopaminergic neurons.

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Neuropharmacology

Background:

  • Dopamine transporter (DAT) regulates dopaminergic neurotransmission by clearing dopamine from synapses.
  • DAT function is potentially modulated by phosphorylation due to identified phosphorylation sites.
  • Previous studies showed protein kinase C (PKC) activation decreases dopamine transport Vmax.

Purpose of the Study:

  • To investigate the in vivo phosphorylation of the rat dopamine transporter.
  • To determine if phorbol ester-induced phosphorylation affects dopamine transporter function.
  • To explore the role of PKC in regulating dopamine transporter activity.

Main Methods:

  • Stable expression of rat dopamine transporter in LLC-PK1 cells.
  • Treatment with phorbol esters (e.g., PMA) to activate PKC.
  • Measurement of dopamine uptake and transporter phosphorylation levels.
  • Analysis of staurosporine sensitivity.

Main Results:

  • Phorbol ester treatment increased dopamine transporter phosphorylation threefold in vivo.
  • Phorbol 12-myristate 13-acetate (PMA) reduced dopamine transport Vmax by 35%.
  • Observed parallels in time course, concentration dependency, and staurosporine sensitivity between phosphorylation and Vmax reduction.

Conclusions:

  • Dopamine transporter phosphorylation, likely mediated by PKC, contributes to decreased transporter function.
  • PKC-mediated phosphorylation may be involved in rapid neuroadaptive processes in dopaminergic neurons.
  • This provides a molecular mechanism for regulating dopamine transporter activity.