MicroRNAs (miRNAs) are small non-coding RNAs that are gaining recognition as important regulators of gene expression with the capability to modulate cellular events. Although miRNAs are involved in a wide range of activities related to the development and differentiation of cells, evidence for miRNAs that control the ethanol-induced differentiation programs of specific neural cell types has been elusive. In this study, we isolated neural stem cells (NSC) from the forebrains of E14 embryos obtained from pregnant C57/ BL6J mice. We used microarray technology to monitor the expression profiles of miRNAs during ethanol-induced differentiation of NSC. Comparison of the miRNA expression profiles of NSC and differentiated NSC showed that the expression levels of 58 miRNAs changed by twofold after differentiation. Among these differentially expressed miRNAs, we confirmed the downregulation of the two miRNAs miR-21 and miR-338-3p and the upregulation of miR-320 by real time RT-PCR. By computational analysis, we identified 48 target genes that are predicted to be regulated by miR-21, miR-338-3p and miR-320 during differentiation. We found that most of these predicted target genes are related to cellular processes, such as apoptosis, neural differentiation, cell cycle, metabolism, cell signaling and development. Finally, our findings may contribute to a better understanding of miRNA-regulated neural differentiation in response to ethanol.
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Temple, S. & Davis, A.A. Isolated rat cortical progenitor cells are maintained in division in vitro by membrane-associated factors. Development 120, 999–1008 (1994).
Temple, S. The development of neural stem cells. Nature 414, 112–117 (2001).
Choi, M.R. et al. Ethanol-induced small heat shock protein genes in the differentiation of mouse embryonic neural stem cells. Arch. Toxicol. 85, 293–304 (2011).
Gong, Z. & Wezeman, F.H. Inhibitory effect of alcohol on osteogenic differentiation in human bone marrowderived mesenchymal stem cells. Alcohol. Clin. Exp. Res. 28, 468–479 (2004).
Jung, K.H. et al. Effects of acute ethanol treatment on NCCIT cells and NCCIT cell-derived embryoid bodies (EBs). Toxicol. In Vitro 24, 1696–1704 (2010).
Li, Z., Lin, H., Zhu, Y., Wang, M. & Luo, J. Disruption of cell cycle kinetics and cyclin-dependent kinase system by ethanol in cultured cerebellar granule progenitors. Brain Res. Dev. Brain Res. 132, 47–58 (2001).
Crews, F.T., Braun, C.J., Hoplight, B., Switzer, R.C., 3rd & Knapp, D.J. Binge ethanol consumption causes differential brain damage in young adolescent rats compared with adult rats. Alcohol. Clin. Exp. Res. 24, 1712–1723 (2000).
Livy, D.J., Maier, S.E. & West, J.R. Fetal alcohol exposure and temporal vulnerability: effects of bingelike alcohol exposure on the ventrolateral nucleus of the thalamus. Alcohol. Clin. Exp. Res. 25, 774–780 (2001).
Hebert, S.S. & De Strooper, B. Alterations of the microRNA network cause neurodegenerative disease. Trends Neurosci. 32, 199–206 (2009).
Kloosterman, W.P. & Plasterk, R.H. The diverse functions of microRNAs in animal development and disease. Dev. Cell 11, 441–450 (2006).
Denli, A.M. & Hannon, G.J. RNAi: an ever-growing puzzle. Trends Biochem Sci. 28, 196–201 (2003).
Bentwich, I. et al. Identification of hundreds of conserved and nonconserved human microRNAs. Nat. Genet. 37, 766–770 (2005).
Croce, C.M. & Calin, G.A. miRNAs, cancer, and stem cell division. Cell 122, 6–7 (2005).
Krichevsky, A.M., King, K.S., Donahue, C.P., Khrapko, K. & Kosik, K.S. A microRNA array reveals extensive regulation of microRNAs during brain development. RNA 9, 1274–1281 (2003).
Conaco, C., Otto, S., Han, J.J. & Mandel, G. Reciprocal actions of REST and a microRNA promote neuronal identity. Proc. Natl. Acad. Sci. USA 103, 2422–2427 (2006).
Schratt, G.M. et al. A brain-specific microRNA regulates dendritic spine development. Nature 439, 283–289 (2006).
Miska, E.A. et al. Microarray analysis of microRNA expression in the developing mammalian brain. Genome Biol. 5, R68 (2004).
Liu, J. et al. Patterns of gene expression in the frontal cortex discriminate alcoholic from nonalcoholic individuals. Neuropsychopharmacology 31, 1574–1582 (2006).
Buller, B. et al. MicroRNA-21 protects neurons from ischemic death. FEBS J. 277, 4299–4307 (2010).
Sathyan, P., Golden, H.B. & Miranda, R.C. Competing interactions between micro-RNAs determine neural progenitor survival and proliferation after ethanol exposure: evidence from an ex vivo model of the fetal cerebral cortical neuroepithelium. J. Neurosci. 27, 8546–8557 (2007).
Przyborski, S.A., Smith, S. & Wood, A. Transcriptional profiling of neuronal differentiation by human embryonal carcinoma stem cells in vitro. Stem Cells 21, 459–471 (2003).
Shimazaki, T., Shingo, T. & Weiss, S. The ciliary neurotrophic factor/leukemia inhibitory factor/gp130 receptor complex operates in the maintenance of mammalian forebrain neural stem cells. J. Neurosci. 21, 7642–7653 (2001).
Song, M.R. & Ghosh, A. FGF2-induced chromatin remodeling regulates CNTF-mediated gene expression and astrocyte differentiation. Nat. Neurosci. 7, 229–235 (2004).
Raghunath, M. et al. A novel kinase, AATYK induces and promotes neuronal differentiation in a human neuroblastoma (SH-SY5Y) cell line. Brain Res. Mol. Brain Res. 77, 151–162 (2000).
Zhao, X. et al. MicroRNA-mediated control of oligodendrocyte differentiation. Neuron 65, 612–626 (2010).
Saba, R., Goodman, C.D., Huzarewich, R.L., Robertson, C. & Booth, S.A. A miRNA signature of prion induced neurodegeneration. PLoS One 3, e3652 (2008).
Schaar, D.G., Medina, D.J., Moore, D.F., Strair, R.K. & Ting, Y. miR-320 targets transferrin receptor 1 (CD71) and inhibits cell proliferation. Exp. Hematol. 37, 245–255 (2009).
Broccardo, C. et al. ABCA2 is a marker of neural progenitors and neuronal subsets in the adult rodent brain. J. Neurochem. 97, 345–355 (2006).
Park, J.H. et al. The characterization of gene expression during mouse neural stem cell differentiation in vitro. Neurosci. Lett. 506, 50–54 (2012).
John, B. et al. Human MicroRNA targets. PLoS Biol. 2, e363 (2004).
Grimson, A. et al. MicroRNA targeting specificity in mammals: determinants beyond seed pairing. Mol. Cell 27, 91–105 (2007).
These authors contributed equally to this work.
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Park, J.H., Halder, D., Choi, M.R. et al. Expression profiles of miRNAs during ethanol-induced differentiation of neural stem cells. BioChip J 6, 73–83 (2012). https://doi.org/10.1007/s13206-012-6110-y
- Neural stem cells
- Neural differentiation
- Microarray analysis