Expression Profiling of Neural Cells Reveals Specific Patterns of Ethanol-Responsive Gene Expression
- Christelle Thibault1,1,
- Chaoqiang Lai2,2,
- Norbert Wilke1,
- Bao Duong1,
- M. Foster Olive1,
- Sajida Rahman1,
- Helin Dong2,
- Clyde W. Hodge1,
- David J. Lockhart2 and
- Michael F. Miles1
- 1The Ernest Gallo Clinic and Research Center and Department of Neurology, University of California, San Francisco, California (C.T., N.W., B.D., M.F.O., S.R., C.W.H., M.F.M.) and2Affymetrix, Inc., Santa Clara, California (C.L., H.D., D.J.L.)
Abstract
Adaptive changes in gene expression are thought to contribute to dependence, addiction and other behavioral responses to chronic ethanol abuse. DNA array studies provide a nonbiased detection of networks of gene expression changes, allowing insight into functional consequences and mechanisms of such molecular responses. We used oligonucleotide arrays to study nearly 6000 genes in human SH-SY5Y neuroblastoma cells exposed to chronic ethanol. A set of 42 genes had consistently increased or decreased mRNA abundance after 3 days of ethanol treatment. Groups of genes related to norepinephrine production, glutathione metabolism, and protection against apoptosis were identified. Genes involved in catecholamine metabolism are of special interest because of the role of this pathway in mediating ethanol withdrawal symptoms (physical dependence). Ethanol treatment elevated dopamine β-hydroxylase (DBH, EC 1.14.17.1) mRNA and protein levels and increased releasable norepinephrine in SH-SY5Y cultures. Acute ethanol also increased DBH mRNA levels in mouse adrenal gland, suggesting in vivo functional consequences for ethanol regulation of DBH. In SH-SY5Y cells, ethanol also decreased mRNA and secreted protein levels for monocyte chemotactic protein 1, an effect that could contribute to the protective role of moderate ethanol consumption in atherosclerotic vascular disease. Finally, we identified a subset of genes similarly regulated by both ethanol and dibutyryl-cAMP treatment in SH-SY5Y cells. This suggests that ethanol and cAMP signaling share mechanistic features in regulating a subset of ethanol-responsive genes. Our findings offer new insights regarding possible molecular mechanisms underlying behavioral responses or medical consequences of ethanol consumption and alcoholism.
Footnotes
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Send reprint requests to: Dr. Michael F. Miles, Ernest Gallo Clinic and Research Center, 5858 Horton St., Suite 200, Emeryville, CA 94608
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↵1 Current address: Institut de Génétique et de Biologie Moléculaire et Cellulaire, B.P. 163, 67404 Illkirch Cedex, France.
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↵2 Current address: Cereon Genomics, 45 Sidney St., Cambridge, MA 02139.
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This work was supported by intramural funding from the Ernest Gallo Clinic and Research Center and by funds provided by the State of California for medical research on alcohol and substance abuse through the University of California, San Francisco.
- Abbreviations:
- DBH
- dopamine β-hydroxylase
- NE
- norepinephrine
- db-cAMP
- dibutyryl-cAMP
- PCR
- polymerase chain reaction
- RT-PCR
- reverse transcriptase PCR
- NET
- sodium-dependent norepinephrine transporter
- DLK1
- δ-like protein 1
- MCP1
- monocyte chemoattractant protein 1
- GAPDH
- glyceraldehyde-3-phosphate dehydrogenase
- ELISA
- enzyme-linked immunosorbent assay
- EST
- expressed sequence tag
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- Received May 16, 2000.
- Accepted August 16, 2000.
- The American Society for Pharmacology and Experimental Therapeutics
