Cell Biochemistry and Biophysics

, Volume 66, Issue 1, pp 131–137

Effects of Homocysteine on ERK Signaling and Cell Proliferation in Fetal Neural Stem Cells In Vitro

  • Hai Yan
  • Xumei Zhang
  • Suhui Luo
  • Huan Liu
  • Xuan Wang
  • Yuxia Gao
  • John X. Wilson
  • Guowei Huang
Original Paper

DOI: 10.1007/s12013-012-9461-z

Cite this article as:
Yan, H., Zhang, X., Luo, S. et al. Cell Biochem Biophys (2013) 66: 131. doi:10.1007/s12013-012-9461-z

Abstract

The aim of the present study was to determine if the excitatory amino acid homocysteine (Hcy) alters ERK signaling and cell proliferation in fetal neural stem cells (NSCs) in vitro. NSCs were isolated from fetal rats and grown in serum-free suspension medium. The cells were identified as NSCs by their expression of immunoreactive Sox2. NSCs were assigned to one of four treatment groups: vehicle control, low-dose Hcy group (Hcy-L, medium contained 30 μmol/L Hcy), middle-dose Hcy group (Hcy-M, 100 μmol/L Hcy) and high-dose Hcy group (Hcy-H, 300 μmol/L Hcy). Cell proliferation was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Protein expression levels of ERK1/2 and phosphorylated ERK1/2 were detected by Western blot. The effects of Hcy on NSC death, including apoptosis, were assessed by using flow cytometry and trypan blue exclusion. The results showed that NSCs grew as neurospheres in the serum-free medium. Hcy decreased ERK1/2 protein phosphorylation and NSC proliferation, but it did not induce cell death or apoptosis within the concentration from 30 to 300 μmol/L. The above results are consistent with the hypothesis that Hcy decreases fetal NSC proliferation by inhibiting ERK signaling.

Keywords

Neural stem cellsHomocysteineProliferationERK signalingIn vitro

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Hai Yan
    • 1
  • Xumei Zhang
    • 1
  • Suhui Luo
    • 1
  • Huan Liu
    • 1
  • Xuan Wang
    • 1
  • Yuxia Gao
    • 2
  • John X. Wilson
    • 3
  • Guowei Huang
    • 1
  1. 1.Department of Nutrition and Food Hygiene, School of Public HealthTianjin Medical UniversityTianjinChina
  2. 2.Department of CardiologyGeneral Hospital of Tianjin Medical UniversityTianjinChina
  3. 3.Department of Exercise and Nutrition Sciences, School of Public Health and Health ProfessionsUniversity at BuffaloBuffaloUSA