PMID- 18579334
OWN - NLM
STAT- MEDLINE
DCOM- 20081016
LR  - 20211020
IS  - 0741-8329 (Print)
IS  - 0741-8329 (Linking)
VI  - 42
IP  - 6
DP  - 2008 Sep
TI  - Ethanol potentiates dopamine uptake and increases cell surface distribution of 
      dopamine transporters expressed in SK-N-SH and HEK-293 cells.
PG  - 499-508
LID - 10.1016/j.alcohol.2008.04.009 [doi]
AB  - Ethanol increases dopaminergic release in the reward and reinforcement areas of 
      the brain. The primary protein responsible for terminating dopamine (DA) 
      neurotransmission is the plasma membrane-bound dopamine transporter (DAT). In 
      vitro electrophysiological and biochemical studies in Xenopus laevis oocytes have 
      previously shown ethanol potentiates DAT function and increases 
      transporter-binding sites. The potentiating effect of ethanol on the transporter 
      is eliminated in Xenopus oocytes by the DAT mutation glycine 130 to threonine. 
      However, ethanol's action on DAT functional regulation has yet to be examined in 
      mammalian cell expression systems. To further understand the molecular mechanisms 
      of ethanol's action on DAT, we determined the direct mechanistic action of 
      short-term (< or =2 h) ethanol exposure on transporter function and cell surface 
      distribution in non-neuronal human embryonic kidney cells-293 (HEK-293) and 
      neuronal SK-N-SH neuroblastoma cells expressing the transporter. Wild-type or 
      G130T mutant DAT were overexpressed in HEK-293 and SK-N-SH cells. Ethanol 
      potentiated DAT mediated [(3)H]DA uptake in a dose (25, 50, 100 mM), but not time 
      dependent manner in cells expressing wild-type DAT. Ethanol-induced potentiation 
      of uptake was significantly reduced in cells expressing the G130T mutant. 
      Analysis of DA uptake kinetic parameters indicates 100-mM ethanol exposure 
      increased [(3)H]DA uptake velocity (V(max)), while affinity for DA (K(m)) 
      remained unchanged. The effect of ethanol on wild-type DAT surface expression was 
      measured by biotinylation cell surface labeling. DAT surface expression increased 
      40%-50% after 1-h, 100-mM ethanol exposure. These studies show ethanol 
      potentiates DAT functional regulation in both neuronal and non-neuronal cells, 
      suggesting a direct mechanistic action of ethanol on transporter trafficking in 
      mammalian systems. Our findings demonstrate ethanol's action on DAT function and 
      regulation is consistent across multiple model systems.
FAU - Riherd, D Nicole
AU  - Riherd DN
AD  - Waggoner Center for Alcohol and Addiction Research, University of Texas at 
      Austin, Austin, TX 78712, USA.
FAU - Galindo, David G
AU  - Galindo DG
FAU - Krause, Lucretia R
AU  - Krause LR
FAU - Mayfield, R Dayne
AU  - Mayfield RD
LA  - eng
GR  - R01 AA014697-02/AA/NIAAA NIH HHS/United States
GR  - R01 AA014697-04/AA/NIAAA NIH HHS/United States
GR  - U01 AA016648-03/AA/NIAAA NIH HHS/United States
GR  - R01 AA014697-01A2/AA/NIAAA NIH HHS/United States
GR  - AA014697/AA/NIAAA NIH HHS/United States
GR  - R01 AA014697/AA/NIAAA NIH HHS/United States
GR  - U01 AA016648/AA/NIAAA NIH HHS/United States
GR  - U01 AA016648-01/AA/NIAAA NIH HHS/United States
GR  - U01 AA016648-02/AA/NIAAA NIH HHS/United States
GR  - R01 AA014697-03/AA/NIAAA NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20080624
PL  - United States
TA  - Alcohol
JT  - Alcohol (Fayetteville, N.Y.)
JID - 8502311
RN  - 0 (Dopamine Plasma Membrane Transport Proteins)
RN  - 3K9958V90M (Ethanol)
RN  - VTD58H1Z2X (Dopamine)
SB  - IM
MH  - Biological Transport, Active/drug effects
MH  - Cell Line
MH  - Cell Line, Tumor
MH  - Dopamine/*metabolism
MH  - Dopamine Plasma Membrane Transport Proteins/*metabolism
MH  - Drug Synergism
MH  - Ethanol/*pharmacology
MH  - Humans
MH  - Kidney/embryology
MH  - Neuroblastoma
PMC - PMC2535921
MID - NIHMS49741
EDAT- 2008/06/27 09:00
MHDA- 2008/10/17 09:00
PMCR- 2009/09/01
CRDT- 2008/06/27 09:00
PHST- 2007/12/13 00:00 [received]
PHST- 2008/04/29 00:00 [revised]
PHST- 2008/04/29 00:00 [accepted]
PHST- 2008/06/27 09:00 [pubmed]
PHST- 2008/10/17 09:00 [medline]
PHST- 2008/06/27 09:00 [entrez]
PHST- 2009/09/01 00:00 [pmc-release]
AID - S0741-8329(08)00218-8 [pii]
AID - 10.1016/j.alcohol.2008.04.009 [doi]
PST - ppublish
SO  - Alcohol. 2008 Sep;42(6):499-508. doi: 10.1016/j.alcohol.2008.04.009. Epub 2008 
      Jun 24.