Ethanol Inhibition of Neural Stem Cell Differentiation Is Reduced by Neurotrophic Factors

Authors


  • This research was supported by the Scientific Research Fund of the Ministry of Education, Science and Culture of Japan.

Reprint requests: Masaru Tateno, Department of Neuropsychiatry, Sapporo Medical University, School of Medicine, South 1 West 16, Chuo-ku, Sapporo, Japan 0608543; Fax: +81-011-644-3041; E-mail: tatema@sapmed.ac.jp.

Abstract

Background: Ethanol exposure during development leads to various forms of neuronal damage. Because neural stem cells (NSCs) play a pivotal role in the development and maturation of the central nervous system, it is important to understand the effect of ethanol on NSC differentiation. In this study, we investigated the effect of ethanol on differentiation of cultured NSCs in the presence and absence of neurotrophic factors.

Methods: NSCs were derived from rat embryos on embryonic day 14. Cells were exposed to ethanol with or without neurotrophic factors, insulin-like growth factor-1 (IGF-1), or brain-derived neurotrophic factor (BDNF). The effect of ethanol on differentiation was quantified by measurement of optical density of each sample following to microtubule-associated protein 2 enzyme-linked immunosorbent assay and counting of the number of microtubule-associated protein 2–positive cells microscopically. In addition, cell viability of cultured cortical neurons that were exposed to similar concentrations of ethanol was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay.

Results: Ethanol (20–100 mM) inhibited NSC differentiation induced by basic fibroblast growth factor removal, whereas those concentrations of ethanol did not affect neuronal survival. Both IGF-1 and BDNF promoted generation of neurons in the absence of ethanol. Moreover, they suppressed the inhibitory effect of ethanol on NSC differentiation.

Conclusions: Ethanol inhibited NSC differentiation at concentrations much lower than what compromised neuronal survival. Ethanol-induced differential inhibition was reduced by both IGF-1 and BDNF. These results suggest that ethanol inhibits stem cell differentiation through alteration of cellular pathways related to neurotrophic factor signaling.

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