PMID- 16181646
OWN - NLM
STAT- MEDLINE
DCOM- 20060310
LR  - 20141120
IS  - 0028-3908 (Print)
IS  - 0028-3908 (Linking)
VI  - 49
IP  - 6
DP  - 2005 Nov
TI  - Differential effects of dopamine and psychoactive drugs on dopamine transporter 
      phosphorylation and regulation.
PG  - 759-68
AB  - The dopamine transporter (DAT) is a phosphoprotein whose activity and 
      phosphorylation state are acutely regulated by both protein kinase C (PKC) and 
      substrate transport. DAT is a major site of action for psychostimulant and 
      therapeutic drugs that either block transport or are transported substrates, but 
      the effects of such drugs on DAT phosphorylation and regulation are not well 
      understood. To examine these issues we subjected rDAT LLC-PK(1) cells to acute in 
      vitro pretreatments with the endogenous, psychostimulant, and therapeutic 
      compounds dopamine (DA), (-)-cocaine, 2 
      beta-carbomethoxy-3beta-(4-fluorophenyl)tropane (beta-CFT), GBR 12909, mazindol, 
      and methylphenidate (MPH), in the presence or absence of the PKC activator 
      phorbol 12 myristate 13 acetate (PMA), followed by analysis of DAT metabolic 
      phosphorylation and transport activity. Basal phosphorylation of DAT was not 
      affected by any of the uptake blockers tested, and PMA-stimulated phosphorylation 
      was not affected by cocaine, beta-CFT, mazindol or MPH, but was strongly 
      suppressed by GBR 12909. Pretreatment of cells with cocaine or MPH had no effect 
      on subsequent DA transport activity or the extent of PMA-induced transport 
      down-regulation, whereas GBR 12909 inhibited PMA-induced DAT internalization. 
      These findings indicate that these DAT phosphorylation and down-regulation 
      properties are unaffected by some classes of uptake blocking drugs, but that 
      differential regulatory effects may be exerted by GBR compounds. Pretreatment of 
      cells with DA had no obvious effect on basal or PMA-stimulated DAT 
      phosphorylation but led to cocaine-blockable transport down-regulation. 
      DA-induced down-regulation was blocked by the PKC inhibitor bisindoylmaleimide I 
      and was not additive with down-regulation induced by PMA, consistent with PKC 
      serving as a common step and point of integration for these DA and PMA induced 
      processes. The results of this study provide information on the potential for 
      endogenous and psychoactive compounds to modulate or be modulated by DAT 
      phosphorylation-mediated regulatory mechanisms that may contribute to drug 
      behavioral or therapeutic properties.
FAU - Gorentla, Balachandra K
AU  - Gorentla BK
AD  - Department of Biochemistry and Molecular Biology, University of North Dakota 
      School of Medicine and Health Sciences, 501 N. Columbia Road, Grand Forks, ND 
      58203, USA.
FAU - Vaughan, Roxanne A
AU  - Vaughan RA
LA  - eng
GR  - R01 DA13147/DA/NIDA NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20050921
PL  - England
TA  - Neuropharmacology
JT  - Neuropharmacology
JID - 0236217
RN  - 0 (Dopamine Plasma Membrane Transport Proteins)
RN  - 0 (Dopamine Uptake Inhibitors)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Phorbol Esters)
RN  - 0 (Phosphorus Isotopes)
RN  - 0 (Piperazines)
RN  - 0 (Psychotropic Drugs)
RN  - 10028-17-8 (Tritium)
RN  - 90X28IKH43 (vanoxerine)
RN  - VTD58H1Z2X (Dopamine)
SB  - IM
MH  - Animals
MH  - Autoradiography/methods
MH  - Biotinylation/methods
MH  - Cell Line
MH  - Dopamine/*pharmacology
MH  - Dopamine Plasma Membrane Transport Proteins/*metabolism
MH  - Dopamine Uptake Inhibitors/pharmacology
MH  - Dose-Response Relationship, Drug
MH  - Drug Interactions
MH  - Enzyme Inhibitors/pharmacology
MH  - Immunoprecipitation/methods
MH  - Phorbol Esters/pharmacology
MH  - Phosphorus Isotopes/metabolism
MH  - Phosphorylation/drug effects
MH  - Piperazines/pharmacology
MH  - Protein Transport/drug effects
MH  - Psychotropic Drugs/*pharmacology
MH  - Swine
MH  - Tritium/metabolism
EDAT- 2005/09/27 09:00
MHDA- 2006/03/11 09:00
CRDT- 2005/09/27 09:00
PHST- 2005/05/17 00:00 [received]
PHST- 2005/07/30 00:00 [revised]
PHST- 2005/08/16 00:00 [accepted]
PHST- 2005/09/27 09:00 [pubmed]
PHST- 2006/03/11 09:00 [medline]
PHST- 2005/09/27 09:00 [entrez]
AID - S0028-3908(05)00315-1 [pii]
AID - 10.1016/j.neuropharm.2005.08.011 [doi]
PST - ppublish
SO  - Neuropharmacology. 2005 Nov;49(6):759-68. doi: 10.1016/j.neuropharm.2005.08.011. 
      Epub 2005 Sep 21.