Altered Expression of 14-3-3 zeta Protein in Spinal Cords of Rat Fetuses with Spina Bifida Aperta

作者：Wu, Li-na; Wei, Xiao-wei; Fan, Yang; Miao, Jia-ning; Wang, Li-li; Zhang, Yi; Wu, Di; Yuan, Zheng-wei^*

来源：PLos One, 2013, 8(8): e70457.

DOI：10.1371/journal.pone.0070457

• NEURAL-TUBE DEFECTS
• DIFFERENTIAL EXPRESSION
• CELL-PROLIFERATION
• DOWN-REGULATION
• OXIDANT STRESS
• BIRTH-DEFECTS
• FETAL RATS
• P53
• SURVIVAL
• APOPTOSIS

Background: A large number of studies have confirmed that excessive apoptosis is one of the reasons for deficient neuronal function in neural tube defects (NTDs). A previous study from our laboratory used 2-D gel electrophoresis to demonstrate that 14-3-3 zeta expression was low in the spinal cords of rat fetuses with spina bifida aperta at embryonic day (E) 17. As a member of the 14-3-3 protein family, 14-3-3 zeta plays a crucial role in the determination of cell fate and anti-apoptotic activity. However, neither the expression of 14-3-3 zeta in defective spinal cords, nor the correlation between 14-3-3 zeta and excessive apoptosis in NTDs has been fully confirmed.
Methodology/Principal Findings: We used immunoblotting and quantitative real-time PCR (qRT-PCR) to quantify the expression of 14-3-3 zeta and double immunofluorescence to visualize 14-3-3 zeta and apoptosis. We found that, compared with controls, 14-3-3 zeta was down-regulated in spina bifida between E12 and E15. Excessive apoptotic cells and low expression of 14-3-3 zeta were observed in the dorsal region of spinal cords with spina bifida during the same time period. To initially explore the molecular mechanisms of apoptosis in NTDs, we investigated the expression of microRNA-7 (miR-7), microRNA-375 (miR-375) and microRNA-451 (miR-451), which are known to down-regulate 14-3-3 zeta in several different cell types. We also investigated the expression of p53, a molecule that is downstream of 14-3-3 zeta and can be down-regulated by it. We discovered that, in contrast to the reduction of 14-3-3 zeta expression, the expression of miR-451, miR-375 and p53 increased in spina bifida rat fetuses.
Conclusions/Significance: These data suggest that the reduced expression of 14-3-3 zeta plays a role in the excessive apoptosis that occurs in spina bifida and may be partly regulated by the over-expression of miR-451 and miR-375, and the consequent up-regulation of p53 might further promote apoptosis in spina bifida.

• 出版日期2013-8-6
• 单位中国医科大学