Neurotransplantation of stem cells genetically modified to express human dopamine transporter reduces alcohol consumption.
BACKGROUND
Regulated neurotransmitter actions in the mammalian central nervous system determine brain function and control peripheral organs and behavior. Although drug-seeking behaviors, including alcohol consumption, depend on central neurotransmission, modification of neurotransmitter actions in specific brain nuclei remains challenging. Herein, we report a novel approach for neurotransmission modification in vivo by transplantation of stem cells engineered to take up the neurotransmitter dopamine (DA) efficiently through the action of the human dopamine transporter (hDAT). As a functional test in mice, we used voluntary alcohol consumption, which is known to release DA in nucleus accumbens (NAC), an event hypothesized to help maintain drug-seeking behavior. We reasoned that reducing extracellular DA levels, by engrafting into NAC DA-sequestering stem cells expressing hDAT, would alter alcohol intake.


METHODS
We have generated a neural stem cell line stably expressing the hDAT. Uptake kinetics of DA were determined to select a clone for transplantation. These genetically modified stem cells (or cells transfected with a construct lacking the hDAT sequence) were transplanted bilaterally into the NAC of wild-type mice trained to consume 10% alcohol in a two-bottle free-choice test for alcohol consumption. Alcohol intake was then ascertained for 1 week after transplantation, and brain sections through the NAC were examined for surviving grafted cells.


RESULTS
Modified stem cells expressed hDAT and uptaken DA selectively via hDAT. Mice accustomed to drinking 10% ethanol by free choice reduced their alcohol consumption after being transplanted with hDAT-expressing stem cells. By contrast, control stem cells lacked that effect. Histologic examination revealed surviving stem cells in the NAC of all engrafted brains.


CONCLUSIONS
Our findings represent proof of principle suggesting that genetically engineered stem cells can be useful for exploring the role of neurotransmitters (or other signaling molecules) in alcohol consumption and potentially in other aspects of brain function.
Department of Medicine, University of Colorado Denver, 12800 East 19th Avenue, Aurora, CO 80045, USA. thangram@hotmail.com
Tom N
TN
Grammatopoulos
Susan M
SM
Jones
Masami
M
Yoshimura
Brian R
BR
Hoover
Mita
M
Das
Evan Y
EY
Snyder
Gaynor A
GA
Larson
Nancy R
NR
Zahniser
Boris
B
Tabakoff
W Michael
WM
Zawada
D000428:Alcohol Drinking / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D004298:Dopamine / Q000378:metabolism; D050483:Dopamine Plasma Membrane Transport Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D058999:Drug-Seeking Behavior / Q000502:physiology*; D005260:Female; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D058953:Neural Stem Cells / Q000378:metabolism* / Q000637:transplantation; D009714:Nucleus Accumbens / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D001616:beta-Galactosidase / Q000378:metabolism
D050483:Dopamine Plasma Membrane Transport Proteins; D001616:beta-Galactosidase; D004298:Dopamine