Ethanol upregulates NMDA receptor subunit gene expression in human embryonic stem cell-derived cortical neurons.
Chronic alcohol consumption may result in sustained gene expression alterations in the brain, leading to alcohol abuse or dependence. Because of ethical concerns of using live human brain cells in research, this hypothesis cannot be tested directly in live human brains. In the present study, we used human embryonic stem cell (hESC)-derived cortical neurons as in vitro cellular models to investigate alcohol-induced expression changes of genes involved in alcohol metabolism (ALDH2), anti-apoptosis (BCL2 and CCND2), neurotransmission (NMDA receptor subunit genes: GRIN1, GRIN2A, GRIN2B, and GRIN2D), calcium channel activity (ITPR2), or transcriptional repression (JARID2). hESCs were differentiated into cortical neurons, which were characterized by immunostaining using antibodies against cortical neuron-specific biomarkers. Ethanol-induced gene expression changes were determined by reverse-transcription quantitative polymerase chain reaction (RT-qPCR). After a 7-day ethanol (50 mM) exposure followed by a 24-hour ethanol withdrawal treatment, five of the above nine genes (including all four NMDA receptor subunit genes) were highly upregulated (GRIN1: 1.93-fold, P = 0.003; GRIN2A: 1.40-fold, P = 0.003; GRIN2B: 1.75-fold, P = 0.002; GRIN2D: 1.86-fold, P = 0.048; BCL2: 1.34-fold, P = 0.031), and the results of GRIN1, GRIN2A, and GRIN2B survived multiple comparison correction. Our findings suggest that alcohol responsive genes, particularly NMDA receptor genes, play an important role in regulating neuronal function and mediating chronic alcohol consumption-induced neuroadaptations.
Department of Genetics, Yale University School of Medicine, New Haven, CT, United States of America.
Yangfei
Y
Xiang
Department of Genetics, Yale University School of Medicine, New Haven, CT, United States of America.
Kun-Yong
KY
Kim
Department of Genetics, Yale University School of Medicine, New Haven, CT, United States of America; Department of Psychiatry, Yale University School of Medicine, New Haven, CT, United States of America; Department of Neurobiology, Yale University School of Medicine, New Haven, CT, United States of America; VA Medical Center, VA Connecticut Healthcare System, West Haven, CT, United States of America.
Joel
J
Gelernter
Department of Genetics, Yale University School of Medicine, New Haven, CT, United States of America.
In-Hyun
IH
Park
Department of Psychiatry, Yale University School of Medicine, New Haven, CT, United States of America; VA Medical Center, VA Connecticut Healthcare System, West Haven, CT, United States of America.
Huiping
H
Zhang
D000428:Alcohol Drinking / Q000235:genetics* / Q000473:pathology; D002454:Cell Differentiation / Q000187:drug effects; D002540:Cerebral Cortex / Q000187:drug effects / Q000473:pathology; D000431:Ethanol / Q000008:administration & dosage*; D005786:Gene Expression Regulation / Q000187:drug effects; D000066449:Human Embryonic Stem Cells / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D009435:Synaptic Transmission / Q000187:drug effects; D015533:Transcriptional Activation / Q000187:drug effects
D000431:Ethanol