Open AccessEthanol Upregulates NMDA Receptor Subunit Gene Expression in Human Embryonic Stem Cell-Derived Cortical Neurons
Author(s) -
Yangfei Xiang,
Kun-Yong Kim,
Joel Gelernter,
InHyun Park,
Huiping Zhang
Publication year - 2015
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0134907
Subject(s) - biology , nmda receptor , immunostaining , gene expression , downregulation and upregulation , microbiology and biotechnology , receptor , gene , genetics , immunohistochemistry , immunology
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.